Background: Thyroid dysfunction plays an important role in the pathology of diabetes-associated cognitive dysfunction (DACD). However, thyroid hormone (TH) signaling and action changes in DACD brains remain unknown. This study evaluated the alternations in TH signaling and action in the brains of DACD mice and explored the beneficial effects of levothyroxine (L-T4) treatment.Methods: KK-Ay mice, serving as a spontaneous type 2 diabetes mellitus model, underwent intragastric administration of 10 ng/g and 20 ng/g of L-T4 solution or normal saline for 8 weeks. Age-matched C57BL/6J mice were used as normal controls. Cognitive and memory functions were examined through the open field and Morris water maze tests. Hippocampal TH signaling and pathogenic status were evaluated. The potential signaling pathways involved in the neuroprotective action of L-T4 were investigated through RNA sequencing and further verified through quantitative real-time PCR (qPCR), Western blotting (WB), immunofluorescence (IF), and fluorescent multiplex immunohistochemistry (mIHC) in vivo and vitro.Results: The expressions of hippocampal TH transporters (Mct8 and Oatp1c1), Dio2, and TH receptor were upregulated, whereas Dio3 as well as the TH-positive regulated genes MBP, Enpp2, and Klf9 were downregulated in DACD mice. Exogenous L-T4 partially alleviated cognitive and memory dysfunction and restored hippocampal neuronal activity by optimizing TH signaling. RNA sequencing provided insights into the role of type I interferon (IFN-I) signaling and necroptosis on the amelioration of hippocampal damage after L-T4 treatment. WB and qPCR further confirmed that the levels of key proteins for IFN-I signaling and necroptosis (p-STAT1, p-STAT2, IRF9, ZBP1, p-RIP3, and p-MLKL) were increased, but largely returned after L-T4 administration in vivo and T3 treatment in vitro. IF and mIHC revealed that IRF9 and p-MLKL colocalized in neurons, but not in astrocytes or microglia, of the hippocampus in DACD mice. The diabetes mellitus group had an increased number of IRF9+ p-MLKL+ NeuN+ cells, which decreased after L-T4 treatment. The elevated IFN-I signaling-mediated necroptosis in HT22 cells was also decreased by T3.Conclusion: We demonstrated abnormal hippocampal TH signaling and action in DACD. Promoting TH action with exogenous L-T4 ameliorated hippocampal impairment through inhibiting IFN-I signaling-induced necroptosis.

The crucial role of plasmid-encoded protein Pgp3 in Chlamydia pathogenesis has been demonstrated in various animal models. Previous studies have revealed that the Pgp3-deficient C. muridarum mutant fails to induce hydrosalpinx after vaginal inoculation in mice. Structural analysis of C. trachomatis Pgp3 trimer has indicated that Trp234 may play a critical role in trimeric crystal packing interactions and that Tyr197 is involved at predominant cation-binding sites. In this study, we constructed C. muridarum transformants harboring Pgp3, Trp234, or Tyr197 point mutations (Pgp3W234A and Pgp3Y197A). C3H/HeJ mice infected with Pgp3W234A mutant failed to induce severe hydrosalpinx in the oviduct tissue, which largely phenocopied the full-length Pgp3-deficient C. muridarum. The Pgp3Y197A variant induced an intermediate severity of pathology. The attenuated pathogenicity caused by the Pgp3W234A mutant may be due to its decreased survival in the lower genital tracts of mice, reduced ascension to the oviduct, and milder induction of inflammatory cell infiltration in the oviduct tissue. Thus, our results point to an important amino acid residue involved in Pgp3 virulence, providing a potential therapeutic target for chlamydial infection.

Oxidative stress plays an important role in acute and chronic inflammatory diseases. However, the clinical treatment options for oxidative stress-related diseases are limited. Nanozyme with excellent reactive oxygen species (ROS) catalytic capacity are promising for preventing ROS-related inflammation. Herein, we designed CeO2:Yb7Tm0.5 with a butterfly-like structure of upconversion nanoparticles (UCNPs) with enzyme-mimicking ability. The CeO2:Yb7Tm0.5 UCNPs have catalase and glutathione peroxidase-mimicking enzyme properties, which can treat acute lung injury (ALI). The CeO2:Yb7Tm(0.5) UCNPs turn blue, green, white, and red under upconversion luminescence with increasing temperature from 298 K to 358 K. Here, the changing of color are examined over the temperature range of in 298 K-373 K in 10 K increments. Maximum absolute and relative sensitivities (S-a and S-r) of 2.3 % K-1 and 2.7 % K-1 are obtained at 298 K and 373 K, respectively. The butterfly-like structure of CeO2:Yb/Tm is promising for biological application, such as accurate cell temperature detection, bio-catalysis diagnostics and ALI treatment.

We have previously shown that circRNAs in host cells are involved in the process of Chlamydia trachomatis infection. In this study we aimed to identify significantly altered circRNAs/lncRNAs/mRNAs in Chlamydia muridarum infected cells and investigate their biological functions in the interaction between Chlamydia muridarum and host cells. For this purpose, circRNA, lncRNA and mRNA expression profiles were screened and identified in HeLa cells with or without Chlamydia muridarum infection by microarray. Bioinformatics analyses including Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) analysis were then carried out and the circRNA-miRNA ceRNA network was constructed. The differentially expressed circRNAs and lncRNAs were selected for validation by RT-qPCR. The results shown that a total of 834 circRNAs, 2149 lncRNAs and 1283 mRNAs were found to be differentially expressed. Enrichment analysis of GO and KEGG showed that the dysregulated genes involved nuclear-transcribed mRNA catabolic process, protein binding, RNA catabolic process and translation, the MAPK signaling pathway, apoptosis, Toll-like receptor signaling pathway, cAMP signaling pathway and Notch signaling pathway may play important roles in Chlamydia infection. Our study provides a systematic outlook on the potential function of non-coding RNAs in the molecular basis of Chlamydia infection.

目的 获得鼠肺炎沙眼衣原体(CM)质粒编码蛋白Pgp3氨基端(n)、羧基端(c)和中间段(m)分别删除的蛋白Pgp3Δn、Pgp3Δm和Pgp3Δc,为近一步研究其功能及沙眼衣原体(C.t)致病机制提供基础.方法 用聚合酶链式反应(PCR)扩增目的基因Pgp3Δn、Pgp3Δm和Pgp3Δc,分别将其定向插入原核表达载体PET-30a(+)中,连接产物转化入大肠埃希菌DH5α、涂板、挑取单克隆提取质粒进行酶切、测序鉴定.鉴定正确的重组质粒分别转化入大肠埃希菌BL21,15℃诱导表达16 h和37℃诱导表达4 h,考马斯亮蓝染色和蛋白质印迹法鉴定目的蛋白.结果 PCR扩增的目的基因Pgp3Δn、Pgp3Δm和Pgp3Δc的长度分别为597、525、324 bp,NdeI和HindⅢ双酶切重组质粒后电泳显示酶切片段大小符合预期,测序结果显示插入片段序列均与基因库中一致;考马斯亮蓝染色和蛋白质印迹显示目的蛋白分子量分别约为22000、18700和12000,3个蛋白片段均在15℃诱导表达16 h的条件下表达量更多.结论 成功表达Pgp3Δn-His、Pgp3Δm-His和Pgp3Δc-His融合蛋白,为进一步研究Pgp3的功能位点和作用机制奠定基础.

[目的]分析沙眼衣原体感染HeLa细胞后宿主细胞中长链非编码RNA(long noncoding RNA,LncRNA)的差异表达谱,并探讨可能的潜在机制.[方法]抽提沙眼衣原体E型标准株感染的HeLa细胞和对照HeLa细胞的RNA,利用LncRNA芯片技术分析并筛选2组之间差异表达的LncRNA.定量PCR验证基因芯片的结果,对差异表达的LncRNA进行GO(gene ontology)功能分析和KEGG通路分析.[结果]共筛选获得2936个差异表达LncRNA,其中上调表达的有1366个,下调的有1570个.GO功能分析和KEGG通路分析结果显示,这些LncRNA与病毒防御反应、炎症反应、细胞内吞、细胞内信号转导、免疫反应相关,并且主要富集在Notch信号通路、肿瘤坏死因子α(tumor necrosis factor α,TNF-α)信号通路、细胞因子-细胞因子受体相互作用等相关通路上.[结论]沙眼衣原体感染宿主细胞后LncRNA的表达有显著差异,提示其可能在沙眼衣原体与宿主细胞的相互作用中发挥重要作用.

While glycogen synthase A deficiency can reduce the growth and proliferation of Chlamydia muridarum, the effect of glycogen synthase A on the pathogenic process of C. muridarum remains unclear. To characterize the effect of glycogen synthase A deficiency on the pathogenicity of C. muridarum in the genital tract, BALB/c mice were intravaginally inoculated with wild-type, plasmid-free and glycogen synthase A-deficient C. muridarum, and the genital tract tissue was isolated to assess the severity of hydrosalpinx and the levels of oviduct dilatation at day 60 after infection. The glycogen storage capacity and in vitro infection ability of different C. muridarum strains were analyzed by periodic acid-Schiff staining and quantification of progeny elementary body(EB) formation. The tissue homogenate was used to determine the recovery of different C. muridarum strains. The results show that glycogen synthase A-deficient C. muridarum induced reduction of hydrosalpinx and attenuated the extent of oviduct dilatation in mice, and exhibited reduced growth and proliferation in the mouse lower genital tract. In addition, glycogen synthase A point mutations at different sites reduced the glycogen storage capacity and in vitro infectivity of C. muridarum to different degrees. Glycogen synthase A deficiency also reduced the host inflammatory reaction and ascending infection of C. muridarum.

Objective:Placental iodide transport is necessary for maintaining an adequate iodide supply to the developing fetus. We hypothesized that compounds from the placental barrier can compensate for decreases in maternal iodine intake and normalize fetal iodine levels. Methods:Pregnant rats administered different amounts of iodine (1.24, 2.5, 5, or 10 mu g/day) were evaluated on gestational day (gd) 16 and 20. The iodine levels in maternal blood, amniotic fluid (AF), and placental tissue were estimated using As-Ce catalytic spectrophotometry. The protein and/or messenger RNA (mRNA) levels of sodium iodide symporter (NIS), pendrin, alpha-smooth muscle actin (alpha-SMA), and CD31 in the placental labyrinth, trophoblast cells isolated using laser capture microdissection (LCM), and/or fetomaternal thyroid were detected using immunoblotting, real-time polymerase chain reaction, and/or immunohistochemistry. Results:When iodine intake was reduced, iodine levels in maternal blood gradually decreased; however, placental iodine levels were not significantly different between groups on gd16 and gd20. Minimal changes were observed in AF iodine levels on gd16, and a mild decreasing trend was observed (iodine dose, 10 to 1.24 mu g/day) on gd20. NIS protein, which was linearly distributed along the basolateral membrane of maternal-fetal thyroid follicles, gradually increased with decreasing iodine levels. Regarding iodine deficiency in the placental labyrinth on gd16 and gd20, pendrin and glycosylated NIS proteins were significantly upregulated in a dose-dependent manner. However, the mRNA levels were unchanged. Furthermore, the conversion of NIS protein from the nonglycosylated to the glycosylated form increased. In trophoblast cells isolated using LCM,PDSmRNA levels increased in the 1.24-mu g/day group on gd16 but notNISmRNA levels. There was a smaller alpha-SMA(+)area in the labyrinth zone on gd16 and gd20; however, the proportional CD31(+)area increased on gd16 and reduced on gd20 with decreased iodine levels. Conclusions:All mechanisms upregulating the expression of iodine transporters and changes in villous stroma and microvessel area in the placental labyrinth can promote iodide transfer from mother to fetus in iodine deficiency, especially before the onset of fetal thyroid function. Compensatory NIS protein regulation in the placenta against decreased iodine intake mainly occurs during translation and glycosylation modification after translation. Pendrin may be more important than NIS in the mediation of placental iodide transport.

Photodynamic therapy (PDT) as a noninvasive technique is widely used to treat cancer diseases due to its low side effects. PDT based on upconversion nanoparticles (UCNPs) improved tissue penetration and photo-stability. However, traditional photosensitizers and UCNPs were difficult to incorporate, which limited the circulation of the UCNPs in blood and decreased the PDT effect. Herein, we designed NaErF4@ZnO UCNPs for potential application in thyroid tumor cell PDT. With ZnO coated on NaErF4, the blue (415 nm), green (525 nm/545 nm) and red (661 nm) upconversion luminescence enhanced compared with that of NaErF4 core nanoparticles. Particularly, the generation of UV upconversion emission by NaErF4 sensitized ZnO, which catalyzed H2O and O2 to produce ROS reactive oxygen species (ROS) to induce papillary thyroid carcinoma (PTC) cell lines BHP 5-16. With 1000 μg mL-1 of NaErF4@ZnO UCNPs, the viability of BHP 5-16 cells decreased to about 41% as measured by CCK8 assay with 980 nm NIR irradiation. Moreover, it was confirmed that NaErF4@ZnO UCNPs had low toxicity for BHP 5-16 cells. All these results indicated that NaErF4@ZnO upconversion nanoparticles were an excellent platform for PDT treatment.

Cerium oxide nanoparticles (NPs) due to their advanced catalytic performance have been widely used to treat oxidative damage. However, Ce2O3 NPs have not been further investigated in the treatment of acute oxidative injury in vivo. It is meaningful to improve the efficiency for treatment of acute oxidative injury with NPs in vivo. In this report, we designed Er3+-doped Ce2O3 (Er/Ce2O3) NPs with a size of 7.9 nm, which were used to treat acute liver injury. Er/Ce2O3 NPs realized high-efficiency catalysis of hydrogen peroxide (H2O2) at room temperature. An acute liver damage model was established through intraperitoneal injection of lipopolysaccharide (LPS) in C57 mice. By analyzing histopathological and biochemical indexes, Er/Ce2O3 NPs showed a significant improvement in LPS-induced acute liver injury. Acute liver oxidative damage can be treated within 24 hours, which proved the high catalytic efficiency of Er/Ce2O3 NPs in vivo. The activities of SOD, GPx and CTA increased and production of ROS decreased with Er/Ce2O3 NP treatment in comparison with LPS-induced injury, indicating that the mechanism of Er/Ce2O3 NPs in the treatment of acute oxidative damage of liver was mainly via catalysis of ROS products. Moreover, the protein expression levels of TNF-α, CD45 and IL-1β in liver decreased in the Er/Ce2O3 NPs-treated group, which indicated that Er/Ce2O3 NPs have the function of anti-inflammation property. Therefore, Er/Ce2O3 NPs can be applied to treat and prevent diseases caused by acute oxidative damage.

Objective To study the effects of selenium combined with vitamin B6 on blood lipid and lipid metabolism in patients with hyperlipidemia,and to explore its regulatory mechanism.Methods A total of 100 patients with hyperlipidemia detected by physical examination during June 2017 to December 2017 were divided into control group and experimental group according to stratified-block random method,with 50 cases in each group.Patients in experimental group took the selenium (50 μg/d) enriched yeast vitamin E tablets and vitamin B6 tablets (10 mg/d).Patients in control group took vitamin E tablets with ordinary yeast.Both of them were treated for 30 days.The routine physical examination (including rounting blood test and blood biochemical examination) was performed on the 0th day and the 30th day,respectively.Serum level of selenium,vitamin B6,total cholesterol,triglyceride,low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol (HDL-C) were measured.On the 30th day of the experiment,the levels of serum 3-hydroxyl-3-methylamyl two coenzyme A reductase (HMGR),liver lipase,lipoprotein lipase (LPL) and total lipase were detected in both groups.The mRNA level of silencing information regulator 1 (SIRT1),peroxisome proliferator-activated receptor αt (PPARαt) and sterol regulatory element binding protein-1c (SREBP-1c) in the whole blood were analyzed by real-time transcription polymerase chain reaction.Results The physical examination indexes of experimental group were not different from those in control group on the 0th day and the 30th day after the intervention (all P ＞ 0.05).Compared with the 0th day,the level of serum albumin in control group and experimental group on the 30th day were decreased (t =5.942,6.162,all P ＜ 0.05).Other physical examination indexes of control group and experimental group were not changed on the 30th day (all P ＞ 0.05).The serum level of total cholesterol and LDL-C on the 30th day in experimental group were lower than those of control group and the 0th day when compared with itself (t =2.960,2.990,all P ＜ 0.05;t =2.190,2.086,all P ＜ 0.05).On the 30th day,compared with control group,the serum level of HMGR was decreased but the liver lipase,LPL and total lipase levels were increased (t =5.839,-2.072,3.115,-2.323,all P＜0.05).The level of SIRT1 and PPARα mRNA in the whole blood of experimental group were higher than those of control group,but the level of SREBP-1 c mRNA was lower than that of control group (t =6.295,7.662,6.857,all P ＜ 0.05).The serum level of selenium and vitamin B6 in experimental group were higher than those of control group at the 30th day and the 0th day when compared with itself (t =-2.606,-2.710,all P ＜0.05;t =-4.296,-3.690,all P ＜ 0.05).Conclusions Moderate supplementation of selenium enriched yeast combined with vitamin B6 can reduce the level of serum total cholesterol and LDL-C among people with hyperlipidemia.The mechanism may be caused by the regulation of the expression of key lipid metabolic regulators such as SIRT1,PPARαt and SREBP-1c,as well as the secretion of lipid metabolism enzymes such as HMGR,liver lipase and LPL.

Objective To investigate the expression of MCT8,DCX,SHH and ARC/ARG3. 1 in brain neurons of neonatal rats exposed to thyroid dysfunction in uterus.Methods Wistar pregnant rats were randomly divided into control group and experimental groups in which rats were drunk water with 1, 3, or 5 ppm propylthiouracil

Non-coding circular RNAs (circRNAs) have been shown to have important roles in many diseases; however, no study has indicated circRNAs are involved in Chlamydia trachomatis infection. In this study, we used circRNA microarray to measure the global circRNA expression profiles in HeLa cells with or without C. trachomatis serovar E (Ct.E) infection. CircRNA/miRNA/mRNA interactions were predicted and bioinformatics analyses were performed. The differentially expressed circRNAs were selected according to our criterion for validation by reverse-transcription and quantitative polymerase chain reaction (RT-qPCR). The mRNA microarray was used to detect the mRNA expression profiles after Ct.E infection. Among 853 differentially expressed circRNAs, 453 were upregulated and 400 were downregulated after Ct.E infection. Target miRNAs and miRNA-targeted mRNAs of these circRNAs were predicted. RT-qPCR analysis indicated hsa circRNA 001226, hsa circRNA 007046 and hsa circRNA 400027 were elevated similar to those determined in the circRNA microarray analysis. The mRNA microarray results showed 915 genes were upregulated and 619 genes were downregulated after Ct.E infection. Thirty-four differentially expressed genes overlapped in the bioinformatics and mRNA microarray results. KEGG pathway analysis revealed several signaling pathways, including endocytosis, MAPK and PI3P-Akt signaling pathways, that were targeted by circRNAs may play important roles in Chlamydia infection. This study provides evidence that circRNAs in host cells are involved in the process of Chlamydia infection.

目的 获取鼠肺炎沙眼衣原体(Chlamydia muridarum)质粒蛋白pgp5的基因及纯化的蛋白并鉴定其免疫原性.方法 设计引物,PCR扩增目的基因,将其定向插入到原核表达载体pET28a中,然后将重组质粒转化入大肠杆菌E.coli DH5α中,并用PCR扩增及序列测定等方法对重组质粒进行鉴定.再将重组质粒转化入感受肽Rosetta(DE3)并诱导表达,用镍柱纯化pgp5-his融合蛋白.用纯化后的目的蛋白免疫新西兰家兔,酶联免疫吸附试验(ELISA)测定抗体效价,Western blot、细胞免疫荧光方法检测抗体与pgp5蛋白的结合.结果 所获得的pgp5基因片段经测序长度为795 bp,检索确认其序列与GeneBank一致.十二烷基硫酸钠一聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹实验均显示获得相对分子质量约29 000的纯化蛋白.ELISA检测多克隆抗体效价达1∶100 000.细胞免疫荧光检测结果显示抗体可与体外培养的鼠肺炎沙眼衣原体特异性结合.结论 成功表达pgp5-his融合蛋白,并制备了高效价、高特异性的抗pgp5抗体,为进一步研究奠定了基础.

Pgp3 consists of globular N- and C-terminal domains connected by a triple-helical coiled-coil middle domain. We demonstrated previously that Pgp3 is required for induction of hydrosalpinx by Chlamydia muridarum. We constructed C. muridarum transformants harboring deletion of the Pgp3 N-terminus (pgp3Δn), C-terminus (pgp3Δc), or middle domain (pgp3Δm). C3H/HeJ and CBA/J mice infected with pgp3Δn or pgp3Δm failed to induce hydrosalpinx in oviduct tissue. However, the pgp3Δc transformant induced mild hydrosalpinx in 20% of C3H/HeJ mice (severity score 0.2 ± 0.6) and in 40% of CBA/J mice (severity score 0.8 ± 1.3). The attenuated pathogenicity of the transformants harboring Pgp3 domain deletions was correlated with impaired in vitro growth and significantly reduced infectivity in the mouse lower genital tract. Moreover, the oviduct tissue of C3H/HeJ and CBA/J mice infected with the Pgp3-domain-deficient transformants displayed less inflammatory cell infiltration. Thus, the structural integrity of plasmid-encoded Pgp3 is essential for induction of hydrosalpinx by C. muridarum.

Chlamydia has a unique intracellular developmental cycle, which has hindered the single protein function study of Chlamydia. Recently developed transformation system of Chlamydia has greatly advanced the chlamydial protein's function research and was used to find that a chlamydial plasmid-encoded Pgp5 protein can down-regulate plasmid-dependent genes. It is assumed, that chlamydial genomic MinD protein has a similar function to Pgp5. However, it is unknown whether MinD protein regulates the same plasmid-dependent genes. We replaced pgp5 gene in the shuttle vector pGFP::CM with minD gene of C. trachomatis (CT0582) or C. muridarum (TC0871). The recombinant plasmid was transformed into plasmid-free organisms-CMUT3 and qRT-PCR was used to detect the transcription level of plasmid-encoded and -dependent genes in these pgp5 deficient organisms. As a readout, GlgA, one of the plasmid-regulated gene products was detected by immunofluorescence assay. After recombination, transformation and plaque purification, the stable transformants CT0582R and TC0871R were generated. In these transformants, the plasmid-dependent genes were up-regulated, alike in the pgp5 premature stop mutant and pgp5 replacement with mCherry mutant. GlgA protein level was also increased in all pgp5 mutants, including CT0582R and TC0871R. Thus, our study showed that genomic MinD protein had different function than Pgp5, which was useful for further understanding the chlamydiae.

Objective To observe the mRNA and protein expression of thyroglobulin (Tg) and thyroid peroxidase (TPO) in each trimester of pregnant and lactating Wistar rats.Methods Ninety-six SPFNAF Wistar rats (84 female and 12 male),weighting 220-260 g were involved.All female Wistar rats were randomly divided into 7 groups according to their body mass via the random number table method:control group,early pregnancy group (7 d),midpregnancy group (14 d),late pregnancy group (21 d),early lactation group (7 d),midlactation group (14 d) and late lactation group (21 d),12 rats in each group.The rats were fed with conventional feed and drank deionized water freely.Female rats of the last 6 groups were mated with male rats.Thyroids were collected on the 7 d,14 d and 21 d of their pregnancy and lactation,respectively.The mRNA expression levels of Tg and TPO were detected by quantitative real-time PCR,and the protein expression levels of Tg and TPO were detected by Western blotting.Results The expression levels of Tg mRNA in thyroid tissue in the control group,early,middle and late pregnancy and early,middle and late lactation (1.05 ± 0.01,3.20 ± 0.23,1.88 ± 0.12,2.69 ± 0.20,1.53 ± 0.19,2.37 ± 0.31,2.23 ± 0.12) were significantly different between groups (F =42.864,P ＜ 0.05),and those of pregnancy and lactation groups were higher than those in the control group (P ＜ 0.05).The expression levels of Tg protein were 0.15 ± 0.01,0.38 ± 0.01,0.32 ± 0.02,0.37 ± 0.01,0.21 ± 0.01,0.35 ± 0.01,0.44 ± 0.01,respectively.The differences between groups were statistically significant (F =232.250,P ＜ 0.05).And those of pregnancy and lactation groups were higher than those in the control group (P ＜ 0.05).The expression levels of TPO mRNA in thyroid tissue in the control group,early,middle and late pregnancy and early,middle and late lactation (0.57 ± 0.01,0.74 ± 0.03,0.78 ± 0.13,1.08 ± 0.10,0.98 ± 0.10,1.00 ± 0.07,0.76 ± 0.05) were significantly different between groups (F =15.448,P ＜ 0.05).And those of pregnancy and lactation groups were higher than those in the control group (P ＜ 0.05).The expression of TPO protein were 0.23 ± 0.01,0.41 ± 0.01,0.72 ± 0.02,0.78 ± 0.01,0.49 ± 0.01,0.52 ± 0.01,0.45 ± 0.02,respectively.The differences between groups were statistically significant (F =563.692,P ＜ 0.05).And those of pregnancy and lactation groups were higher than those in the control group (P ＜ 0.05).Conclusions The mRNA and protein expression levels of TPO and Tg have increased in pregnant and lactating rats.This performance may be raleted to thyroid hormone deficiency and mild hypothyroidism.

Chlamydia trachomatis infection is one of the most prevalent sexually transmitted diseases. Our research pertains to the inhibitory effect and molecular mechanism of the chlamydiaphage capsid protein VP1 on the growth of Chlamydia trachomatis. In this research, the capsid protein VP1 of the guinea-pig conjunctivitis chlamydiaphage phiCPG1 was expressed, purified and identified, and then, it was applied to the cultivation of different serovars of Chlamydia trachomatis and Chlamydia psittaci. The inhibitory effect was observed in each serovar of Chlamydia trachomatis (D, E, F, G, H, I, K, and L2) and Chlamydia psittaci inoculated with VP1 protein. The inhibition affection of VP1 on the growth of Chlamydia trachomatis was caused by the changes of expressions of some related proteins including 36 proteins up-regulated and 81 proteins down-regulated in the development cycle of Ct through the label-free test, and the transcription levels of these proteins, including Hc1, pmpD, and MOMP, were confirmed by RT-PCR. It provides information that is essential for understanding the mechanism of chlamydiaphage capsid protein VP1 on chlamydia and a new direction for further clinical treatment of chlamydial infection.

Objective To observe and compare the different orthotopic models of papillary thyroid cancer ( PTC) cell lines of RET/PTC1 rearrangement and BRAFV600E mutation in nude mice. Methods Human PTC cell lines TPC-1, BHP5-16 and BHP2-7 were used. The genotypes of RET/PTC1 rearrangement and BRAFV600E mutation were determined by realtime-PCR and DNA sequencing analysis. The cells(2×105) were injected into the thyroid gland of nude mice. The nude mice were executed at 4th, 12th week, and then their thyroid tumors were removed and weighed. The levels of thyroid hormone were detected using chemiluminescent immunoassay. Results Both TPC-1 and BHP2-7 cells were identified as RET/PTC1 rearrangement by real time-PCR, and the expression of RET/PTC1 rearrangement in BHP2-7 cell was higher than that of TPC-1 cell. BRAFV600E mutation was found in BHP5-16 cell by DNA sequencing analysis, but was not found in TPC-1 and BHP2-7 cells. There were different characteristics in three orthotopic nude model groups. Tumorigenic rates of TPC-1 and BHP5-16 groups were 100%, but the growth of tumor was more rapid in BHP5-16 group than that in TPC-1 group, with more weight tumor. The changes of thyroid hormone levels in BHP5-16 group and TPC-1 group were the same, which were normal at 4th week and sharply decreased at 12 th week(P<0. 05). However, the tumorigenic rate of BHP2-7 group was only 6. 25%. Compared with normal control group, there was no statistical difference in the levels of thyroid hormone in BHP2-7group(P>0. 05). Conclusions It showed difference in the orthotopic models of PTC cell lines of RET/PTC1 rearrangement and BRAFV600E mutation in nude mice. BRAFV600E mutation has obvious impacts on increasing tumorigenic rate and promotion of tumor growth in the orthotopic model. It should not be ignored that advanced thyroid tumor will lead to the destruction of thyroid function.

Transmembrane protein transporters mediate cellular uptake and efflux of thyroid hormones.Monocarboxylate transporter 8 (MCT8) plays an essential role in the supply of T3 to neurons in the central nervous system.So far, MCT8 is the only one with specific clinical significance and can importantlly transport THs into brain.MCT8 (encoded gene is SLC16A2) mutations lead to Allan-Herndon-Dudley Syndrome (AHDS) with severe neurological impairment and altered concentrations of thyroid hormones.The endocrine component in Mct8-deficiency mice is likely to be similar to the humans'.However, unlike in humans with an MCT8 deficiency, there is almost not neurological impairment in these mice.After all, deep insight for clinical features with MCT8 mutations can be partly explained and the pathophysiological role of thyroid hormone transporters can be partially identified in Mct8-deficiency mice.