The anti-tumor clinical effect of traditional Chinese medicine gecko is very prominent. A novel anti-tumor protein component F2 was isolated and purified from geckos by our group, and the aim of this study was to investigate its antitumor effects in vitro and in vivo and its mechanism of action by inhibiting angiogenesis. The proliferation of cells was determined by colony formation assay and MTT assay. The apoptosis of A549 cells was detected by flow cytometry, and the gene expression changes of cells was analyzed by RNA-seq technology. An H22 tumor-bearing mouse model was prepared, the tumor tissue was analyzed by CD31 immunohistochemical staining. The expression levels of target proteins were detected by Western blot. The proliferation of tumor cells A549 and BEL-7402 was significantly inhibited by F2, but the growth of normal cells L02 was not affected. F2 induces apoptosis in A549 cells and inhibits its migration. RNA-seq results showed that F2 significantly inhibited MAPK signaling pathway in A549 and BEL-7402 cells, and the related genes MEK2 and ERK were significantly down-regulated. The zebrafish thrombus model test showed that the extract of gecko has a significant inhibitory effect on thrombus formation in zebrafish. F2 showed a tumor inhibition rate of 42.24% in mice. A decrease in blood vessels around the tumor and in the tumor tissue was observed. The treatment resulted in a significant down-regulation of VEGF and p-ERK expressions, while a significant up-regulation of p-JNK. In conclusion, F2 has significant antitumor activity. One of its mechanisms of action is to suppress angiogenesis in tumor tissue and surrounding tissues by inhibiting the VEGF/ERK/MAP signaling pathway.

Background and aim Transdermal drug delivery has been used in traditional Chinese medicine for thousands of years and is worth further developing. The purpose of this study was to investigate the preparation method of Tubiechong gel plaster (TGP) and evaluate its activity to promote fracture healing. Experimental procedure Using rat skin as a barrier, and the human umbilical vein endothelial cells (HUVECs) proliferation promotion rate of the receiving solution as the evaluation index, the in vitro permeability of the plaster was studied by Franz diffusion cell method. A modified Einhorn method was used to model rat tibia fractures. The bone healing process was monitored by radiography. Bone development and angiogenesis were assessed by Safranin O Fast Green staining and CD31 immunohistochemistry, respectively. Serum bone metabolites and VEGF levels were determined by ELISA method. Results and Conclusion The optimal dose of azone for TGP is 2%. The transdermal penetration equation for this plaster is consistent with the Ritger-Peppas equation. The prepared plaster was stable for at least 3 months. X-ray images showed that the fracture healing was promoted by TGP in a dose-related form at all periods. Serum BALP, OC, CTX-I, and VEGF levels also indicated better fracture healing after treatment. TGP significantly stimulated angiogenesis and bone growth at the fracture site on days 7,14 and 21, and the effect of promoting bone growth was shown on the third day. These data prove that the prepared Tubiechong gel plasters can promote fracture healing by promoting angiogenesis.

One of the factors that predispose to fractures is liver damage. Interestingly, fractures are sometimes accompanied by abnormal liver function. Polyene phosphatidylcholine (PPC) is an important liver repair drug. We wondered if PPC had a role in promoting fracture healing. A rat model of tibial fracture was developed using the modified Einhorn model method. X-rays were used to detect the progression of fracture healing. Progress of ossification and angiogenesis at the fracture site were analyzed by Safranin O/fast green staining and CD31 immunohistochemistry. To investigate whether PPC has a direct angiogenesis effect, HUVECs were used. We performed MTT, wound healing, Transwell migration, and tube formation assays. Finally, RT-qPCR and Western blot analysis were used to study the underlying mechanism. The results showed that PPC significantly shortened the apparent recovery time of mobility in rats. PPC treatment significantly promoted the formation of cartilage callus, endochondral ossification, and angiogenesis at the fracture site. In vitro, PPC promoted the proliferative viability of HUVECs, their ability to heal wounds, and their ability to penetrate membranes in the Transwell apparatus and increased the tube formation of cells. The transcription of VEGFA, VEGFR2, PLCγ, RAS, ERK1/2 and MEK1/2 was significantly up regulated by PPC. Further, the protein level results demonstrated a significant increase in the expression of VEGFA, VEGFR2, MEK1/2, and ERK1/2 proteins. In conclusion, our findings suggest that PPC promotes angiogenesis by activating the VEGFA/VEGFR2 and downstream signaling pathway, thereby accelerating fracture healing.

ObjectiveTo determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability.MethodsA modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels.ResultsTu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment.ConclusionTF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Ethnopharmacological relevance: The external use of traditional Chinese medicine (TCM) to treat fractures has a long history of clinical application and theoretical basis, and is also one of the characteristic treatment methods of TCM with significant efficacy and many advantages. Among the commonly used external Chinese medicines, Tubiechong is noteworthy.Aim of the study: To elucidate whether local patching of Tubiechong can promote fracture healing and explore its mechanism of action. Materials and methods: A rat tibia fracture model was constructed by the modified Einhorn modeling method. Xray films were taken to evaluate the progress of fracture healing. Serum bone alkaline phosphatase (BALP), osteocalcin (BGP) and the C-terminal content of collagen type I (CTX-I) were analyzed by ELISA. CD31 immunohistochemistry was used to evaluate angiogenesis in the tibia segment. The effects of Tubiechong decoction (TD) on HUVEC proliferation, migration and invasion were detected by MTT assay, wound healing assay and Transwell migration assay, respectively. RNA-seq was performed to identify differentially expressed genes (DEGs). Enrichment of functions and signaling pathway analysis were performed based on the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Quantitative real time polymerase chain reaction (qRT-PCR) was used to study gene expression levels. Western blotting (WB) was used to detect the expression of relevant regulatory proteins.Results: The healing time of rat tibia fractures in the three TD dose groups was shortened. The serum levels of BALP, BGP and CTX- I in the TD-treated group were higher than those in the NC group. The X-ray results showed that on the 7th day after surgery, the fracture healing degree of the high-dose TD group was significantly better than that of the NC group, and the fracture healing degrees of each TD treatment group were significantly higher than those of the NC group on the 14th, 17th, and 21st days after the operation. The CD31 immunohistochemistry results showed that the number of blood vessels and the vascular area in the TD treatment group were higher than those in the NC group. In vitro, TD promoted the proliferation, wound healing and migration of HUVECs. GO analysis of transcriptome sequencing results showed that TD significantly altered the expression of genes related to cell growth, metabolism, and motility. According to KEGG annotations, VEGFA was upregulated. Eight DEGs were enriched in the VEGFA-VEGFR2 signaling pathway, of which six were upregulated. KEGG signaling pathway analysis showed that the most abundant DEGs were in mitogen-activated protein kinase (MAPK) signaling pathway. qRT-PCR showed that VEGFA gene expression in HUVECs was 7.8 times that of the control group after 1 mg/mL TD treatment for 24 h, and WB experiments showed that its protein expression was 3 times that of the control group. WB results showed that the phosphorylated ERK gene was highly expressed, while the expression levels of phosphorylated P38 and phosphorylated JNK protein remained unchanged.Conclusion: Tubechong patching therapy promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE:Tubiechong comprises mainly Eupolyphaga and Steleophaga is widely distributed in China. It has been used in the traditional medicine systems in Asian countries specially in China，Japan and Singapore for thousand years.AIM OF THE REVIEW:The aim of this work is to review the scientific work about Tubiechong regarding their ethnomedicinal uses, bioactive chemical constituents and pharmacological activities.MATERIALS AND METHODS:Relevant literature of Tubiechong was collected for its traditional uses, pharmacological activities, and bioactive compounds released from inception until May 2022. The online databases such as Web of Science, PubMed, Google Scholar, Science Direct, Scopus, SciFinder Scholar, Springer Link, China National Knowledge Infrastructure (CNKI), Wanfang Data, and VIP database were used as electronic search engines for articles with the various specific keywords. Additionally, references from ancient texts and local information such as PhD and MSc theses, books, and Chinese journals were also included.RESULTS:The clinical researches have revealed that Tubiechong alone has been successfully used to treat bone disease, ache, sprain, herpes zoster, paronychia and so on. Tubichong's main clinical application is to form formulations with other herbs. The most widely used 34 kinds of Chinese patent medicine containing Tubiechong were included in Chinese Pharmacopoeia (2020 Edition) for the treatment of traumatic injury, low back pain, cardiovascular disease, tumors or mass and nodule, cervical spondylopathy, osteoarthritis and psoriasis. Its other derived formulas have been used in the clinical treatment of various diseases, such as blood stasis, hepatic cirrhosis, cyclomastopathy, chronic active hepatitis, nephropathy, gynaecopathia, cancer diseases. To date, the bioactive substances reported are limited to protein and peptides, fatty acids, polysaccharides and alkaloids from Eupolyphaga sinensis Walker. So far, the pharmacological activities of Tubiechong and its various extracts have been evaluated, including anticoagulant and antithrombotic, anticancer, bone repair, immunomodulation, analgesia, antioxidant, antihyperlipidemic, antimicrobial and protective and repair functions for damage to the liver, heart, brain and skin. As an edible insect, its safety has also been confirmed by acute toxicity tests and 30-day feeding trials.CONCLUSION:Tubiechong is an important insect medicine with the effect of promoting blood circulation and removing blood stasis, which has been used in traditional Chinese medicine for thousands of years for the treatment of trauma and abdominal lumps, and has now been clinically extended to the treatment of a variety of diseases. Its multiple pharmacological activities indicate that it has great potential for development and application. However, its chemical constituents with pharmacological activity require further excavation and detailed study. In addition, the in-depth molecular pharmacological mechanisms deserve further explanation.

目的:制备性状良好的土元凝胶贴膏.方法:采用单因素试验结合Box-Behnken设计响应面法,以初黏力、持黏力和剥离强度为评价指标,对土元凝胶贴膏基质配比进行优选,同时使用正交设计法筛选最佳载药量.结果:最佳基质配比为聚丙烯酸钠(NP-700)10.00 g、甘羟铝0.40 g、酒石酸0.32 g、甘油36.00 g、高岭土8.00 g、水24.00 g,载药10％土元冻干粉.结论:优选得到的凝胶贴膏黏性适中、剥离强度小,可用于后续土元凝胶贴膏的开发与应用.

目的 探究人参皂苷Rb2在体内调控破骨细胞的作用及机制.方法 通过去势建立小鼠骨质疏松模型,分为4组(假手术组,去势组,低剂量组,高剂量组),腹腔注射不同浓度Rb2溶液[低剂量组5 mg/(kg·d),高剂量组20 mg/(kg·d),假手术组、去势组注射蒸馏水],常规饲养12周.使用Elisa试剂盒检测血清破骨细胞活性指标抗酒石酸酸性磷酸酶(tartrate-resistant factor acid phosphatase,TRAP)、Ⅰ型胶原C端肽(C-terminal telopeptide-Ⅰ,CTX-1)水平,股骨远端TRAP染色,从髓腔中提取和培养原代破骨细胞,TRAP染色并计数检测破骨细胞分化情况,Western-blot技术检测原代破骨细胞核因子κB (nuclear factor kappa-B,NF-κB)p65蛋白及自噬相关分子自噬标志轻链3(autophagy marker light chain 3,LC3)、哺乳动物雷帕霉素蛋白(mammalian target of rapamycin,mTOR)、5'-磷酸腺苷活化蛋白激酶(adenosine 5'-monophosphateactivated protein kinase,AMPK)、磷酸化腺苷酸活化蛋白激酶(phosphorylated adenosine monophosphate activated protein kinase,pAMPK)蛋白表达水平.结果 去势小鼠血清TRAP(64.10±1.18) pg/ml、CTX-1(70.43±1.71)ng/ml浓度最高,其余各组血清指标明显降低,高剂量组尤甚[分别为(44.62±2.02) pg/ml和(49.55±2.72)ng/ml](P＜0.01).去势组股骨远端TRAP染色阳性细胞密集,阳性区域最大,随着Rb2浓度增加,TRAP染色阳性区域面积减少(P＜0.05).去势组多核破骨细胞数量最多(152.00±18.55/孔),低剂量组(118.40±18.46/孔)和高剂量组(99.60±13.89/孔).TRAP染色阳性细胞数量均有不同程度减少,低剂量组最少(84.00±15.23/孔),各组P＜0.01.与去势组相比,各组NF-κB在细胞质增加(P＜0.05),在细胞核减少(P＜0.01);各组mTOR表达减少,AMPK、pAMPK表达增加,LC3 Ⅱ/LC3 Ⅰ比值增高(P＜0.05).结论 人参皂苷Rb2在体内通过抑制NF-κB信号通路以及调控mTOR介导的自噬信号通路抑制破骨细胞.

鳄鱼是现存最古老的爬行动物,国内人工养殖技术已经成熟,鳄鱼血有很高的经济价值及药用价值.本试验研究了人工养殖暹罗鳄血清、血浆及白细胞提取液体外对甲型流感病毒、单纯疱疹病毒、乙肝病毒的作用.结果表明,人工养殖暹罗鳄血清对甲型流感病毒的半数抑制浓度(IC50)为原液的0.103％,乙肝病毒HBsAg、HBeAg的IC50分别为原液的0.312％和0.54％,对单纯疱疹病毒无抑制作用;血浆对甲型流感病毒的IC50为原液的0.086％,对乙肝病毒HBsAg、HBeAg的IC50分别为原液的0.36％和0.13％,对单纯疱疹病毒无抑制作用;白细胞提取液对甲型流感病毒及单纯疱疹病毒无抑制作用,对乙肝病毒HBsAg的IC50为原液的0.002％,对乙肝病毒HBeAg无抑制作用.结果说明,暹罗鳄血清及血浆中存在某些可以体外抑制流感病毒的物质.

To determine the contents of EGF,IGF-1 and NGF in different sections of sika deer antler and explore the effects of hot processing treatment.The velvet antler was treated by traditional hot processed method.The contents of the soluble proteins in the velvet antler were measured by Bradford method.The contents of the growth factors in the upper,middle and lower sections of the velvet antler were determined by ELISA.Results:In terms of fresh velvet antler,the EGF,IGF-1,NGF levels were 2.45ng/mL,18.18ng/mL and 14.59pg/mL in the upper,0.94ng/mL,7.79ng/mL and 7.95pg/mL in the middle respectively;in the lower section,no EGF was detected and the IGF-1,NGF levels were 1.19ng/mL and 2.47pg/mL respectively.The data for the processing antler in the upper section were 0.28ng/mL,1.05ng/mL and 3.99pg/mL for EGF,IGF-1 and NGF respectively;in the middle section,the NGF levels were 2.0 pg/mL and no EGF,IGF-1 were detected;in the lower section,none of the three growth factors was detected.Conclusion:Deer velvet antler growth factors' levels were decreased from the top to the base.The traditional hot processing treatment significantly lowered the growth factor contents in various sections of the velvet antler.

Objective: To investigate the anti-tumor effect of macromolecular fractions of fresh gecko (M-AG) in vivo and their differentiation-inducing activity in Bel-7402 cells in vitro. Methods: An H22 hepatocarcinoma-bearing mouse model was used to evaluate the anti-tumor activity of M-AG samples. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was applied to analyze cell viability. Cell morphology was observed by phase contrast microscopy. The quantity of the alpha-fetoprotein was detected by a radioimmunoassay. Chromatometry was used to assay the albumin quantity. Activities of alkaline phosphatase and γ-glutamyl trans-peptidase were measured by biochemical methods. Finally, western blotting was applied to assess proteins in the mitogen-activated protein kinase (MAPK) signaling pathway. Results: Macromolecular fractions of fresh gecko exerted a significant anti-tumor effect in mice. The inhibition rate of tumor growth was 63% in the moderate M-AG dose group. Cells treated with M-AG displayed a differentiated state. The treatment lowered alpha-fetoprotein secretion and significantly decreased the activities of γ-glutamyl trans-peptidase and alkaline phosphatase in Bel-7402 cells. In contrast, M-AG increased the amount of albumin in the cell culture medium. All biochemical indices demonstrated that M-AG induced Bel-7402 cell differentiation. Western blotting showed no changes in the quantities of extracellular signal-regulated kinase (ERK) 1/2, p38 MAPK, or c-Jun N-terminal protein kinase 1/2. However, M-AG significantly activated the phosphorylation of ERK1/2 in a dose-dependent manner. In addition, M-AG had no significant influence on the expression of nuclear factor-kappa B. Conclusion: Macromolecular fractions of fresh gecko has an anti-tumor activity in H22 hepatocarcinoma-bearing mice in vivo and inhibits Bel-7402 cell proliferation in vitro by inducing cell differentiation related to activation of ERK1/2.

Objective To investigate effects and mechanism of ginsenoside Rb2 on osteoclasts in vitro. Methods The mouse osteoclast precursor cells RAW 264.7 were cultivated to induce osteoclasts. Rb2 ( 0.1 μM, 1 μM, 10 μM ) was respectively added in the culture solution. CCK-8 was used for the detection of drug toxicity. The morphology and count of osteoclasts were observed by TRAP staining. The expression of osteoclast specific gene mRNA, such as TRAP and NFATc1, was detected by RT-PCR. Western blot technique was used to detect the expression levels of LC3 I, LC3 II and mTOR proteins. Results Compared with the control group ( 168.2 ± 22.6 ), the number of osteoclasts decreased significantly: group 0.1 μM ( 131.0 ± 16.3 ), P = 0.018; group 1 μM ( 98.8 ± 17.9 ), P<0.01;group 10 μM(85.8 ± 17.3),P<0.01.The expression of TRAP and NFATc1 osteoclast specific gene mRNA decreased significantly, the ratio of LC3, II / LC3 and I increased significantly, and the amount of mTOR decreased, P< 0.01. Conclusions Rb2 may affects the expression of osteoclast genes by autophagy pathway to reduce the osteoclast formation. mTOR pathway plays an important role in this process.

Objective To investigate the effect of sensory and motor nerve homogenates at different concentrations on the proliferation and osteogenesis differentiation of bone marrow mesenchymal stem cells (BMSCs) in rats.Methods The saphenous nerve and the muscle branch of the sciatic nerve in rats were extracted surgically as sensory and motor nerve tissues,respectively.The primary nerve homogenates (10 mg/mL) were prepared as per 10 mg tissue with 1 mL osteoblast inducing conditional media,and 10 times diluted after filtration purification to prepare sensory and motor nerve homogenates at concentration gradients of 1.0,0.1,0.01,0.001 and 0.0001 mg/mL.Cultivation GFP ± rat pups BMSCs in vitro were trained to P3 generation.The experiment was carried out in 3 groups.The sensory and motor nerve homogenates of 500 μL at the above 6 concentration gradients were added during cultivation respectively in the sensory nerve group (n =18) and the motor nerve group(n =18) while 500 μL of osteoblast inducing conditional media was added in the control group(n =3).Cell proliferation quantity detection and alkaline phosphatase (ALP) staining were used to assess the proliferation and differentiation of BMSCs after 14 days.Results According to the results of CCK-8,the cellular absorbance values at concentrations of 1.0 and 0.1 mg/mL homogenate in the sensory nerve group (1.957 ±0.065 and 1.751±0.073) were significantly greater than in the control group (1.145±0.087) while the cellular absorbance value at concentration of 10.0 mg/mL homogenate in the motor nerve group (0.304 ± 0.619) was significandy smaller than in the control group (1.145 ± 0.087) (P ＜ 0.05).According to the ALP staining,the amounts of cellular calcium nodules in the sensory and motor nerve groups (2.667 ± 0.816 and 3.000 ± 0.632,respectively) were significantly smaller than in the control group (11.833 ± 1.471) (P ＜ 0.05).Conclusion Sensory nerve homogenate is different from motor nerve homogenate in that it may promote proliferation of BMSCs and inhibit osteogenesis differentiation of BMSCs in a certain rage of concentrations.

Objective To investigate the effects of structural changes of scaffolds on the proliferation of nerve cells.Methods The primary cultured and purified Schwann cells(SCs)were included from the sciatic and brachial plexus nerve of 1-3 days oldfetal SD rats.The purified SCs were inoculated on the scaffolds with different pore sizes by the drip method to determine the proliferation of SCs. Morphology and molecular biology were used for the examination.Results The cell proliferation in the large aperture group was significantly better than that in the small aperture group.The CCK-8 cell proliferation curve showed thatthe cell proliferation at 96 h in the large aperture group(0.43±0.01)was significantly higher than that in the small aperture group(0.38±0.01)(P<0.05).The findings of ELISA showed that the expression of nerve growth factor in the large aperture group was significantly higher than that in the small aperture group(P<0.05).Conclusion Although the small aperture scaffoldmaterial stimulates the proliferation of cells at the early stage,the biological behavior of SCs was inhibited. Although the early cell base of the large aperture scaffoldmaterial is insufficient,the three-dimensionalspatial structure of the scaffold material is enough to accommodate cells,so that the number of cells gradually increases. Thus,the different three-dimensional spatial structure of the scaffold material may directly affect the proliferation of nerve cells on the scaffold material.

Liposomes have successfully been used for decades to encapsulate and protect drugs that are prone to deactivation in the body. The present study aimed to demonstrate the use of liposomes to encapsulate cordycepin, an adenosine analog that quickly loses its activity in vivo. The cordycepin-loaded liposomes were prepared by the ammonium sulfate gradient approach, and its in vitro and in vivo antitumour activities were evaluated using BEL-7402 cells and hepatocellular carcinoma H22 transplanted tumors, respectively. An MTT assay was used to observe the cytotoxicity of cells treated with cordycepin and cordycepin-loaded liposomes in vitro. High-content screening (HSC) was carried out using Hoechst 33342 to detect apoptotic cells and the ratio of cells in different cell cycle stages. The data demonstrated that both the cordycepin and the cordycepin-loaded liposomes resulted in clear cytotoxicity with IC50 values of 18.97 and 29.39 mu g/mL, respectively. The latter showed significantly strong inhibitory effects on H22 tumor growth in mice, while the former did not show any inhibitory effects on tumor growth. In addition, the HSC assay showed that the cordycepin-loaded liposomes resulted in a higher rate of apoptosis than the cordycepin alone in BEL-7402 cells. Further data analysis revealed that the cells treated with cordycepin-loaded liposomes were predominately arrested at the G2/M phase (p < 0.05), while those treated with cordycepin alone were arrested in the G0/G1 phase (p < 0.05). In conclusion, these results suggest that liposomes can enhance and maintain the in vivo anti-tumor activity of cordycepin.

OBJECTIVE: To assess the acute organ toxicity of Strychnos nux-vomica with zebrafish model visually.METHODS: To assess acute toxicity, we. initially determined the lethal concentration after Strychnos nux-vomica treatment for 24 h. Zebrafish was treated with five concentrations <= LC10 for 24 h, and the effects of Strychnos nux-vomica on morphology, function of heart, central nervous system, liver, kidney and organ-specific cell death were assessed. Next, we assessed the reversibility of toxic effect.RESULTS: Strychnos nux-vomica has an effect, on the different organs of zebrafish, including heart, central nervous system, liver, and kidney, and cadio-toxEcity induced by Strychnos nux-vomica was reversible to some extent.CONCLUSION: Zebrafish model is suitable for confirming the toxic target organs for Chinese traditional medicine. (C) 2016 JTCM. All rights reserved.

目的:探讨交感神经分泌的神经肽Y(NPY)和感觉神经分泌的钙基因相关肽(CGRP)在体内骨折愈合的不同阶段的变化及意义.方法:选择6-8月龄的雄性大鼠,建立大鼠的股骨闭合骨折模型,术后2、4、8、12周取材.进行扫描电镜,免疫组织荧光染色和血清Elisa检测.结果:①骨折愈合不同时期感觉神经肽类物质CGRP和交感神经肽类物质NPY都有表达,且其含量有先增加后减少的趋势,并在骨折后8周含量达到最高.②骨折愈合不同阶段的大鼠血清感觉神经肽类物质CGRP和交感神经肽类物质NPY均呈上升趋势,差异有统计学意义(P＜0.05),且NPY的含量比CGRP的含量高.骨折后2-4周,CGRP含量增加较快;骨折后4-8周NPY含量增加较快.结论:骨折愈合的不同阶段,感觉神经肽类物质CGRP和交感神经肽类物质NPY含量先升后降,对不同阶段的骨形成及骨吸收产生影响.

Hepatocellular carcinoma is one of the leading causes of malignancy-related death in China. Its therapy in clinics is a big challenge. Ginsenoside Rh2 is one of the most notable cancer-preventing components from red ginseng and it has been reported that ginsenoside Rh2 exhibited potent cytotoxicity against human hepatoma cells. Rh2 exists as two different stereoisomeric forms, (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2. Previous reports showed that the Rh2 epimers demonstrated different pharmacological activities and only (20S)-ginsenoside Rh2 showed potent proliferation inhibition on cancer cells in vitro. However, the in vivo anti-hepatoma activity of (20R)-ginsenoside Rh2 and (20S)-ginsenoside Rh2 has not been reported yet. This work assessed and compared the anti-hepatoma activities of (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 using H22 a hepatoma-bearing mouse model in vivo. In addition, hematoxylin and eosin staining, the deoxynucleotidyl transferase dUTP nick-end labeling assay, and the semiquantitative reverse transcriptase polymerase chain reaction method were used to further study the apoptosis of the tumors. The results showed that both (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 suppressed the growth of H22 transplanted tumors in vivo, and the highest inhibition rate could be up to 42.2 and 46.8%, respectively (p < 0.05). Further, hematoxylin/eosin staining and the deoxynucleotidyl transferase dUTP nick-end labeling assay indicated that both (20R)-ginsenoside Rh2 and (20S)-ginsenoside Rh2 could induce H22 hepatoma tumor cell apoptosis, with apoptosis indexes of 3.87%, and 3.80%, respectively (p < 0.05). Moreover, this effect was accompanied by downregulating the level of Bcl-2 mRNA. In conclusion, both (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 can suppress the growth of H22 hepatomas without causing severe side effects, and this effect is associated with the induction of apoptosis.

This study aimed to find better methylesterification method for analyzing the fatty acid composition of the tomato ,Hami melon ,grapes or pomegranate seed oils .The seed oils of the four fruits were methylated by acid-catalyzed ,alkali-catalyzed ,acid-alkali combination method ,re-spectively ,and the fatty acids composition were analyzed by gas chromatography-mass spectrometry .The results showed that there were 12 ,11 ,12 or 12 kinds of fatty acids identified in the tomato ,Hami melon ,grape or pomegranate seed oil ,respectively ,by KOH-methanol solution method;8 ,8 ,6 or 5 kinds of fatty acids by H2 SO4 -methanol solution method;11 ,15 ,14 or 11 kinds of fatty acids by acid-alkali combination method . These data suggest that KOH-methanol solution method is suitable for methylesterification of tomato seed oil or pomegranate seed oil;KOH-methanol solution and H2SO4 -methanol solution combination method are fitting for Hami melon or grape seed oil .