O bjective:To evaluate the effectoflosartan,an angiotein Ⅱ receptorantagonist,on large arterialbuffering function in elderly patients with hypertension.M ethods: One hundred and eightpatients,of them 85 m ale and 23 fem ale,m edian aged (66.48 ± 5.16)years were given losartan 50 m g per day.The carotid- fem oralartey pulse wave velocity (PW V)m easurem ents were undergone in the subjects before treatm entand 3 m onthsaftera treatm ent.R esults:Three m onthsaftertreatm ent,the systolic and diastolic blood pressure values were significantly reduced (P=0.002 2,P=0.008 1),and carotid-fem oralartery PW V was significantly decreased [(13.60 ± 2.31)m/svs ( 11.89 ± 2.11 ) m/s , P=0.041 6]. C onclusion:Losartan isan effective anti鄄hypertensive drug, and can im prove large artery buffering function in patientswith essentialhypertension.
Objective: To synthesize polyoxyethylene-β-cyclodextrin (PE-β-CD) and study its hemolysis in vitro and its effect on the solubilization of quercetin in water. Methods: β-cyclodextrin was used to react on epoxy ethane in alkali solution to obtain polyoxyethylene-β-cyclodextrin which was identified by IR spectrophotometry and 13C-NMR spectral analysis. The hemolysis was deteated by means of red blood cells isolated from whole rabbit blood and the effect of PE-β-CD on solubilization of quercetin in water was studied by UV spectrophotometry. Result: The hemolytic potential of PE-β-CD was much smaller than that of β-CD, and also smaller than that of HP-β-CD. PE-β-CD also exhibited a remarkable effect on solubilization of quercetin in water. Conclusion: PE-β-CD may well be applied as a novel auxiliary material for injection.
Objective:To investigate the morphology and viability of granulosa cells in primordial follicles from fresh and cryopreserved human ovarian tissues.Methods:Two or three ovarian cortexes were randomly selected for histological examination from freshly obtained human ovarian tissue.The remaining ovarian material was incubated with collagenase type I and deoxyribonuclease I or performed to store in liquid nitrogen.After partial disaggregation,follicles were isolated by manual dissection.A double-staining method was used to assess granulose cells and oocyte viability.Results:In the fresh group,the viable percent of follicles with damaged granulosa cells and oocytes in tissue section were 12.4% and 20.6% compared with 23.3% and 24.4% in the frozen-thawed group by histological analysis.However,by double staining,72.7%,3.6% of freshly and 79.5%,2.5% of preservedly non-damaged granulosa cells and 50% of live granulosa cells in primordial follicles were viable by live-dead staining.When oocyte viability was considered alone,85.5% and 86.1% of the cells were viable in fresh and frozen-thawed follicles,respectively.Conclusion:Granulosa cells,not oocytes of human primordial follicles are affected by the slowly-freezing procedure.
Objective: To evaluate the effect of heparanase antisense oligodeoxynucleotide (ASOND) on the expression of CD44v6 in human gastric carcinoma cell line SGC-7901. Methods: Human gastric adenocarcinoma cell line SGC-7901 was transfected with liposome-mediated heparanase AS-OND. The expression of heparanase mRNA was detected after transfection by semi-quantitative total RNAs RT-PCR. The change of CD44v6 was observed by using immunocytochemistry. Results: The heparanase mRNA expression and CD44v6 expression of SGC-7901 cells after treated with AS-ODN 48 hours at different final concentrations were significantly decreased compared with those in the controls (P0.01). The expression rate of CD44v6 was 69.4% before transfection, and 56.5%, 54.1%, 46.5% and 41.3% after treated with 0.1 μmol/L, 0.2 μmol/L, 0.4 μmol/L and 0.8 μmol/L AS-ODN of the final concentrations, respectively. Conclusion: Heparanase AS-ODN has a significant inhibitory effect on CD44v6 expression in human gastric carcinoma cell line SGC-7901 in a dose-dependent manner.
Objective: To investigate the toxic effects of herpes simplex virus-thymidine kinase (HSV-TK) gene system combining with recombinant IFNα-2a (rIFNα-2a) on human prostate cancer cell line PC-3m. Methods: HSV-TK gene was transfected into PC-3m cells through retrovirus vector, reverse transcription-polymerase chain reaction(RT-PCR) was used to demonstrate the successful transfection and the transcription of HSV-TK gene. Microculture tetrajolium test(MTT) method, flow cytometry (FCM) were used to determine the killing effect of Ganciclovir (GCV), rIFN-α2a, or both on PC-3m cells being transfected with HSV-TK gene. Results: Significant cytotoxic activities were found by GCV on HSV-TK gene-transduced PC-3m cells, the 50% inhibitory concentration (IC50) was 8.34μg/ml, but the bystander effect was modest. The simultaneous treatment of GCV and rIFN-α2a could significantly strengthen the bystander effect, coefficient of two drug interaction (CDI) was less than 0.7. Conclusion: HSV-TK gene therapy combining with rIFN-α2a results in synergistic toxicity on PC-3m cells in vitro.
Objective:To explore the humoral immunological response induced by classⅠand class Ⅱ anti-human leucocyte antigen(HLA)antibodies after implantation of conduit valved homograft(CVH).Methods:From October 2001 to October 2003,20 patients suffered from complex congenital heart disease(CHD)underwent transplantations of cryopreserved homografts artery.The other 20 complex CHDs underwent corrected operation without CVH implantations.The serums were taken and the frequency of panel reactive antibodies directed against HLA classⅠ(HLA-A,B,and C)and class Ⅱ(HLA-DR/DQ)alloantigens were detected by LAT ELISA kit before and 1 month,3 months,1year after operation.Results:The antibodies were both negative before and after operation in control patients.In CVH implanted patients,the frequency of panel reactive antibodies against HLA classⅠwas(9.28±6.64)% at 1 month,(62.14±11.95)% at 3 months and(66.79±13.42)% at 1 year after CVH implantation;that against HLA classⅡ was(22.92±15.74)% at 1 month,(41.67±18.73)% at 3 months and(52.92±20.01)% at 1 year after operation.And the panel reactive frequency of classⅠand class Ⅱ anti-HLA antibodies presented an upward trend within 1 year and a negative correlation with age.Age negatively correlated with the degree of elevation of classⅠand class Ⅱ anti-HLA antibodies.Conclusion:Implantation of CVH can induce a marked humoral immunological response involved classⅠand class Ⅱ anti-HLA antibodies;this response is stronger in young than in eld.
Objective: To explore the relations between neuron apoptosis and hippocampus sclerosis of human temporal lobe epilepsy. Methods: The specimens were obtained from 24 patients with temporal lobe epilepsy, investigated neuron apoptosis by light microscopy, electron microscopy and TdT-mediated dUTP-biotin nick end labeling (TUNEL) method, and determined the expression of Bcl-2, Bax and P53 by immunohistochemistry. Results: The evidence of neuron apoptosis was not found by light microscopy, electron microscopy and TUNEL method in control group. But some early signs of neuron apoptosis in 3 patients(3/8) were found by electron microscopy in epilepsy group. Unlike that in normal adult brain, Bcl-2 immunoreactivity was observed in some neurons in epilepsy group (P0.01); compared with normal group, Bax immunoreactivity was not increased in epilepsy group(P0.05); P53 immunoreactivity was found in some neurons and reactive astroglias in epilepsy group(P0.05). Conclusion: Neuron apoptosis is involved in hippocampus sclerosis of human epilepsy. Bcl-2 and P53 protein may play a role in this process.
Objective: To investigate the role of mutated mismatch repair gene hMLH1 and hMSH2 in the carcinogenesis of sporadic colorectal carcinoma (SCC). Methods: hMSH2,hMLH1 and p53 gene in normal and tumor tissue in 45 SCC specimens were detected by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) EB staining. Results: The mutation rate of hMLH1,hMSH2 and p53 expression was 4.44%(2/45),13.33%(6/45)and 48.89%(22/45)in SCC respectively. The frequency of mutated mmr (hMLH1 and hMSH2) in mutated p53 patients was 27.27%,while it was 8.69% in un-mutated p53 patients (P0.05). Conclusion: There is a subset of SCC displaying MMR gene defect,in which the hMLH1 gene plays more important role in SCC than hMSH2 does. Furthermore,there is a closely relationship between mutation of MMR gene and mutation of p53 gene.
Objective: To study the mechanism of spermatogenesis and its influence factors, the spermatogonia had to be separated and purified to provide technological groundwork and cells source. Method: Continuous enzymatic digestion was used to prepare spermatogenic cell suspension of male mouse of 7~8 days; Percoll density gradient centrifugation combined with plating culture method was used to isolate and purify spermatogonia;alkaline phosphotase of the cells suspension drop and the testis tissues of the same time were detected. Results: Spermatogonia were mainly distributed in Percoll gradient between 45%~55%,purity of spermatogonia was 75.2% after being purified. Conclusion: Percoll density gradient centrifugation combined with continuous enzymatic digestion and plating culture method can isolate and purify spermatogonia from mouse 7~8 days postnatal.
Objective: To explore the indications and the therapeutic efficacy of mastoid and middle ear surgery in chronic otitis media with effusion. Methods: A retrospective study on 30 patients (36 ears) with chronic secretory otitis media was performed. All the 36 ears underwent intact canal wall masto-atticotomy and exploratory tympanotomy, of them, 28 were performed ventilation tube insertion, and 5 ears had ossicular reconstruction simultaneously. The histopathological examination showed inflammative granuloma(n=28) and cholesterol granuloma (n=8). Results: The tympanogram results showed that 27 ears were type A, 8 ears were type C, and 1 ear was type B. 27 ears (27/28) were extubated at 1 to 3 months after surgery with healed drums, the other 1 ear had effusion in middle ear recurred and had the ventilation tube to now. Postoperatively followed up for 0.5-3 years, all had hearing improvement, 31 ears (86.11%)had their average speech threshold within 30dB. 2 ears transferred from type C to type A. Conclusion: The mastoid and middle ear surgery should be performed if the chronic secretory otitis midia can not be cured by several kinds of treatment, because of its completely clearance of the diseased tissue, reconstruction of the ventilation function of the tympanic cavity, tympanic sinus and mastoid process, and prevention of recurrence.
Objective: To evaluate the immunosuppressive effect of Cyclosporin A(CsA) on aortic valve allograft durability and its prevention on calcification. Methods: Aortic valve homograft was heterotopically allografted onto abdominal aorta from Sprague-Dawley (SD) rats to Wistar rats. The rats were divided into 3 groups, test group treated with CsA (0.15 mg/kg), isogeneic group and control group. The rats were sacrificed in batches at 2, 4, 8,12 and 16 weeks postoperatively. Blood samples were obtained for accessing the expression of CD25, CD71, and AVH were obtained for accessing the content of calcium. At the same time, the changes of endotheliocyte and the smooth muscle cells were observed under transmission electron microscope. Results: The expressions of CD25 and CD71 in control group were much higher in 2~4 weeks after operation than those in other times. The expressions of CD25 and CD71 in test group and isogeneic group were much lower than those of control group at early stage of transplantation. The content of calcium in control group increased at 4 weeks after operation and reached the highest level at 12 weeks after operation. The content of calcium in isogenenic group had no difference with that of pre -transplant. The content of calcium in test group was lower than that of control group at 4-16 weeks after operation. Conclusion: CsA treatment can prevent calcification after aortic valve homograft transplant.
Objective: To investigate the changes of serum levels of TNF-α,STNFRI,IL-10 and TGF-β1 in CHF patients with different classes of cardiac function. Methods: The severe levels of serum TNF-α,STNFRI,IL-10 and TGF-β1 were determined by ELISA in 56 patients with moderate to congestive heart failure (CHF) and 30 healthy subjects. Results: Compared with matched controls,CHF patients had significantly elevated serum levels of TNF-α,STNFRI,IL-10 and TGF-β1 (P0.01). TNFα/STNFRI and TNFα/IL-10 were also notably elevated in these patients (P0.05,P0.01). Conclusion: There is enhanced activation of inflammatory cytokines in CHF and the levels of these cytokines are closely related with cardiac function of CHF.
Objective: To investigate the effects of interleukin-1beta (IL-1β) on the expression of RANTES in endometrial cells and evaluate the relationship between proinflammtory cytokine IL-1β and chemokine RANTES and the importance of IL-1β in the development of endometriosis. Methods: The endometrial stromal cells of endometriosis cultured within 3～5 passages were exposed to various concentrations of IL-1β. The expression of RANTES mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The amount of RANTES protein was assessed by enzyme-linked immunosorbent assay (ELISA). Results: ① Expression of RANTES mRNA was positive in unstimulated endometrial stromal cells. However, after stromal cells were incubated with IL-1β, the intensity of RANTES mRNA expression was obviously increased and demonstrated a dose- and time-dependent manner. The expression of RANTES mRNA markedly increased in the presence of IL-1β at 0.1ng/ml for 4 hours, and reached the peak at 1.0 ng/ml for 8 hours, but decreased progressively by time . ② RANTES protein was detected but tiny in culture supernatant of which the cells were not treated. Increase of RANTES protein in culture supernatant was observed following stimulation with 0.1ng/ml IL-1β for 4 hours, and the maximal stimulation effect of RANTES protein in supernatant happened in the presence of 1.0ng/ml IL-1β for 12 hours. Conclusions: IL-1β induces endometrial stromal cell of endometriosis to express RANTES not only at transcription level but also at post-transcription level in a dose- and time-dependent manner.
Objective: To investigate the expression and subcellular localization of the spike (S) protein of SARS-CoV fused with tandem affinity purification (TAP) tag in Vero cells and to serve the identification of the cellular binding proteins of S protein. Methods: The S gene with Kozak sequence was cloned by PCR from the plasmid pGEM-4Z-S, then inserted into the vector pcDNA3.1(-)with the TAP tag comprised calmodulin binding peptide(CBP) and protein A motifs. The expression and subcellular localization of the fusion protein S-CBP-ProteinA[S-TAP(C)] in Vero cells were respectively analyzed by indirect immunofluorescence assay(IFA) and Western-blotting assay(WB) following the transfection with the construct and co-infection with the recombinant vaccinia virus vTF7-3 expressing T7 RNA polymerase. Results: The eukaryotic expression vector pcDNA3.1-S-TAP(C) was constructed correctly. After the transient transfection with the plasmid, expression of the fusion protein S-TAP(C) was detectable by both IFA and WB. The fusion protein was located in the cell compartment of plasma and membranes. The expression level was dramatically enhanced by co-infection with vTF7-3 before transfection. Conclusion: The fusion protein comprised the spike protein and the TAP tag is successfully expressed in Vero cells with a desired subcellular location and can be used in the future screening of cellular binding proteins.
Objective: To study the mRNA expressions and levels of gelatinase-A and B in pneumococcal meningitis. Methods: Thirty-six wistar rats were infected intracisternally with streptococcus pneumoniae, and their pathological changes were observed. The mRNA expressions of gelatinase-A and B in brain parenchyma were verified by RT-PCR analysis, and gelatinolytic activities of gelatinase-A and B in cerebrial spinal fluid(CSF) were determined by gelatin zymographic analysis. Results: Acute inflammation was observed in subarachnoid space. The mRNA expression and relative gelatinolytic activity of gelatinase-B in pneumococcal group were significantly increased (1.04±0.34;1 554.2±264.5, respectively), but there were no changes in the expression and relative gelatinolytic activity of gelatinase-A (P0.05). Conclusion: There is an up-regulation of gelatinase-B which may contribute to the pathogenesis of pneumococcal meningitis.
Objective: To observe whether Ca2+-activated Cl- channels play a role in norepinphrine (NE)-induced contraction of rat tail artery. Methods: The tail artery was isolated from the anesthetized Wistar rats. Under a dissecting microscope the artery was cut helically into strip and then mounted vertically on a stainless steel holder in an organ chamber containing PSS aerated with a mixture of 95% O2-5% CO2. The isometric contraction was recorded. The contractile reactivity of the vessel strip to different chemicals was expressed as a percentage of 100 mmol/L KCl-induced contraction of the same strip. Results: ①The Cl- channel blocker-niflumic acid (NFA) produced a dose-dependent inhibitory effect on NE-induced contraction of the rat tail artery strip. The Ca2+ channel blocker-Verapamil (Vera) had an inhibitory effect on NE-induced contraction of the artery strip. The inhibitory effects of NFA and Vera were additive and coordinated. ② Decreasing the extracellular Cl- concentration increased the contractile response to NE significantly,which was still inhibited by NFA. Conclusion: The Ca2+-activated chloride channel activity contributes to NE-induced contraction of the rat tail artery.
Objective: To evaluate the relationships of serum ferritin(SF) with arteriosclerosis and other cardiovascular risk factors. Methods: IMT and plaque were detected in 88 patients with high risks of arteriosclerosis by ultrasound of high resolution power. The Crouse was scored. The level of SF was determined by microwell ferritin EIA. Patients were separated to normal and abnormal group according to: (1) whether the IMT≥0.8 mm or not; (2) whether the plaque existed or not. Results: The SF was significantly higher in abnormal group than that in normal group(P0.01), patients were significantly older in the abnormal group than that in the normal group(P0.01). The average value of carotid IMT(IMTmean), the peak value of carotid IMT(IMTmax) and the Crouse score of patients with SF≥300 mg/L was significantly higher than those with SF300 mg/L(P0.01, P0.01, P0.01). The level of SF was positive correlated with average value of carotid IMT(IMTmean), the peak value of carotid IMT(IMTmax) and the Crouse score on linear correlation and regression analysis(r=0.294,P0.01;r=0.238,P0.05; r=0.391, P0.01). The level of SF was not correlated with age, blood pressure, lipids, blood glucose and CRP on multiple stepwise regression analysis. The level of SF was significantly higher in patients with smoking than that in patients without smoking(P0.05). Conclusion: Iron plays an important role in the development of arteriosclerosis.
Objective: To evaluate the application of magnetic resonance imaging 、MRI) in detecting the depth of myometrial invasion in patients with endometrial carcinoma. Method: MRI was performed in 51 patients with endometrial carcinoma to diagnose the depth of myometrial invasion. Results: MRI made right diagnosis in all 51 patients with endometrial carcinoma before operation. The total accurate rate of MRI in detecting the depth of myometrial invasion was 86.27% 、44/51). The accurate rate in detecting superficial and deep myometrial invasion was 88.46%、23/26) and 88.23%、15/17) respectively. The accurate rate of dynamic gadolinium-enhanced MR imaging was 90.5% 、19/21). Conclusion: MRI, especially dynamic MRI, plays an important role in the diagnosis of endometrial carcinoma and in judging the depth of myometrial invasion, and should be an useful way in preoperative diagnosis of endometrial carcinoma.
