The effect of empirical antifungal treatment on the diagnostic sensitivity of galactomannan in bronchoalveolar lavage fluid in chronic obstructive pulmonary disease patients combined with invasive pulmonary aspergillosis / 中华老年医学杂志

Objective:To investigate the effect of empirical antifungal treatment on the diagnostic sensitivity of galactomannan(GM)in bronchoalveolar lavage(BALF)in chronic obstructive pulmonary disease(COPD)patients combined with invasive pulmonary aspergillosis(IPA).Methods:COPD patients with IPA were enrolled between January 2015 and January 2019 as research objects.Patients who were treated with antifungal drugs prior to bronchoscopy were considered as the empirical group, and other patients were considered as the diagnosis-driven group.The results of BALF GM were compared between the two groups.Results:A total of 66 COPD patients with IPA were enrolled in this study, with 5 cases confirmed and 61 cases clinically diagnosed.Of them, 17 cases were in the empirical group and 49 in the diagnosis-driven group.There was no significant difference in the sensitivity of blood GM and microbiological examination between the two groups( χ2=0.248 and 0.379, P=0.619 and 0.538). With BALF GM 0.5 as a cutoff value, the sensitivity of BALF GM was slightly lower in the empirical group than in the diagnosis-driven group but with no significant difference(88.2%, 95% CI: 62.3%-97.9% vs.93.9%, 95% CI: 82.1%-98.4%, χ2=0.051, P=0.821). However, with BALF GM 1.0 as a cutoff value, the sensitivity decreased greatly in the empirical group compared with the diagnosis-driven group(52.9%, 95% CI: 28.5%-76.1% vs.80.6%, 95% CI: 67.4%-90.8%, χ2=4.036, P=0.045). Logistic regression analysis showed that after adjusting for mechanical ventilation( OR=0.807, 95% CI: 0.215-3.026, P=0.750)and use of semisynthetic penicillins( OR=0.498, 95% CI: 0.140-1.776, P=0.283), the false negative rate of BALF GM was associated with empirical antifungal therapy( OR=0.243, 95% CI: 0.068-0.949, P=0.030). Conclusions:Empirical antifungal therapy prior to bronchoscopy can decrease the diagnostic sensitivity of BALF GM in COPD patients with IPA.

hCD47 induces immune tolerance of human macrophages to porcine endothelial cells / 器官移植

Objective To investigate the effect of human CD47 (hCD47) in inducing the immune tolerance of human macrophages to porcine endothelial cells. Methods The porcine iliac endothelial cell (PIEC) transfected with pCDH-hCD47-FLAG plasmid was assigned into the pCDH-hCD47 group, PIEC transfected with pCDH-FLAG empty vector plasmid was assigned into the pCDH group, PIEC transfected with hCD47-dN was assigned into the pCDH-hCD47-dN group and human umbilical vein endothelial cell (HUVEC) was assigned into the positive control group. The cells were co-cultured with human macrophages to detect and analyze the phosphorylation of signal regulatory protein α (SIRPα) and the killing effect of human macrophages on PIEC. Furthermore, porcine arteriae endothelial cell (PAEC) was isolated from GT－/－ and GT－/－/ hCD 47 gene editing pigs to analyze the phosphorylation of SIRPα and the killing effect of human macrophages on PAEC. Results The pCDH group cells could not induce the phosphorylation of SIRPα, whereas the pCDH-hCD47 group cells could activate the phosphorylation of SIRPα after 10 min co-culture with human macrophages, and the degree of phosphorylation of SIRPα was increased with the prolongation of the co-culture time. The pCDH-hCD47-dN group cells failed to activate the phosphorylation of SIRPα. Human macrophages exerted significant effect on killing the pCDH group cells. The pCDH-hCD47 group cells could evidently inhibit the killing effect of human macrophages (P < 0.05), whereas the pCDH-hCD47-dN cells failed to suppress the killing effect of human macrophages. GT－/－-PAEC could not activate the phosphorylation of SIRPα after co-culture with human macrophages. However, GT－/－/hCD47-PAEC significantly activated the phosphorylation of SIRPα after co-culture with human macrophages. Human macrophages exerted significant killing effect on GT－/－-PAEC, and GT－/－/hCD47-PAEC could obviously inhibit the killing effect of human macrophages (P < 0.05). Conclusions The expression of hCD47 in the porcine endothelial cells can inhibit the killing effect of human macrophages on endothelial cells by activating the phosphorylation of SIRPα.

Comparison of blood glucose-lowering function of transplant islets between subcutaneous adipose tissues of inguinal region and renal capsule in mice / 器官移植

Objective To compare the effect of transplant islets between the subcutaneous inguinal white adipose tissues and renal capsule in the treatment of type 1 diabetes mellitus in mouse models. Methods The mice with type 1 diabetes mellitus undergoing islet transplantation were divided into the white adipose group (n=10) and renal capsule group (n=10). The islets were isolated, purified and transplanted to the subcutaneous white adipose tissues of inguinal region and renal capsule. The random blood glucose level and glucose tolerance function of the recipient mice in two groups were continuously monitored after operation. Islet grafts of the surviving recipient mice were harvested at postoperative 100 d for histopathological examination. Results In the white adipose group, the blood glucose levels of 6 recipient mice were restored to normal at 1 month after transplantation, whereas the blood glucose levels of the other 4 recipient mice were high, which died before the end of monitoring. In the renal capsule group, the blood glucose levels of 10 recipient mice returned to normal within 10 d after transplantation. Islet grafts of the recipient mice in two groups could lower the blood glucose levels, whereas the islet grafts in the white adipose group required a longer time to exert the effect. The glucose tolerance function of the mice in the renal capsule group was significantly better than that of those in white adipose group (P < 0.05). Histopathological examination demonstrated that the insulin of the islet grafts was normally expressed in two groups. Conclusions The islets transplanted into the subcutaneous white adipose tissues of inguinal region can play an effective role in regulating the changes of blood glucose level. Although the blood glucose-lowering function is slightly weaker than that of the islets graft in the renal capsule, it has multiple advantages resembling the ideal islet transplantation sites, which is a promising replacement site for islet transplantation.

Study of immunoregulatory effect of sirolimus on xenotransplantaion with arterial patch / 器官移植

Objective To investigate the immunoregulatory effect of sirolimus on the xenotransplantation with arterial patch. Methods The xenotransplantation of arterial grafts was taken from the wild-type Bama pigs to cynomolgus monkeys. The peripheral blood mononuclear cells of recipient monkeys at 14 days after xenotransplantation (POD14) were selected as subjects. Dimethyl sulphoxide (DMSO) was used in the control group (volume ratio of 1:1 000) and sirolimus was administered in the sirolimus experimental group (final concentration of 0.1 μmol/L and 0.5 μmol/L). The cells were cultured for 1.0 and 5.5 d, respectively. The activity of POD14 cells was evaluated. The DMSO control and sirolimus experimental groups (final concentration of 0.1 μmol/L) were established and cultured for 5.5 d. The quantity of T and B cells in POD14 cells was counted and the expression levels of cytokines and messenger RNA (mRNA) were quantitatively measured. Results Compared with the DMSO control group, the activity of POD14 cells was significantly decreased after sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 1.0 d (P<0.01-0.001). After sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 5.5 d, the activity of POD14 cells was significantly decreased (both P<0.001). Compared with the DMSO control group, the quantity of CD3+CD4+T cells and CD3+CD8+T cells in POD14 cells was significantly reduced after sirolimus treatment at a final concentration 0.1 μmol/L (P<0.05-0.01), whereas the quantity of CD3-CD20+B cells was considerably elevated (P<0.01). Compared with DMSO control group, the levels of interferon(IFN)-γ, interleukin(IL)-2, IL-4, IL-5 and IL-6 in the sirolimus experimental group were significantly down-regulated (P<0.05-0.001). The expression levels of IFN-γ, tumor necrosis factor(TNF)-α, IL-2, IL-4, IL-5 and IL-6 mRNA were significantly down-regulated (P<0.05-0.001). Conclusions Sirolimus inhibits the proliferation of POD14 cells in the recipient monkeys after xenotransplantation with arterial patch. The underlying mechanism is that the sirolimus can reduce the quantity of T cells and suppress the expression and secretion of immune rejection-related cytokines.

Detection methods of non-Gal xenoantibody in human serum / 器官移植

Objective To investigate the optimal condition for the detection of anti-non-galactose (Gal) xenoantigen and antibody in human serum.Mehtods Peripheral blood mononuclear cell (PBMC) obtained from Wuzhishan miniature pig models with α-1,3-galactosyltransferase gene knockout (GTKO) were used as target cells,mixed and incubated with healthy human serum of different concentrations (4.8％,16.7％ and 100％) for 0.5,1.0,2.0,3.0 and 6.0 h,respectively.The abilities of PBMC to bind with IgM and IgG were detected by flow cytometry.Results At the serum concentration of 16.7％,the ability ofnon-Gal IgM to bind with PBMC was significantly enhanced from 0.5 h to 3.0 h incubation (P＜0.01),whereas no statistical significance was noted in terms of IgG (P＞0.05).Increasing serum concentration could also enhance the ability of non-Gal IgM to bind with PBMC.At the serum concentration of 100％ and incubation for 3 h,the ability of IgM to bind with PBMC was the highest among all groups (P＜0.01).At the serum concentration of 100％ and incubation for 6 h,the ability of IgG to bind with PBMC was significantly enhanced (P＜0.05).Prolonging incubation time and increasing serum concentration did not affect the activity of PBMC.Conclusions The optimal condition for detection of anti-non-Gal xenoantigen and antibody is determined.A quantity of 1×105 PBMC from pig should be incubated with 100％ human serum for 3 h for detection of IgM level,or incubated with 100％ human serum for 6 h for measurement of IgG level.This optimized condition contributes to screening the donor pigs which lowly express non-Gal antigen.

Monitoring of immune rejection after abdominal aortic patch suture in cynomolgus monkeys / 器官移植

To establish a platform to monitor the immune rejection after abdominal aortic patch suture in a xenotransplantation model.Methods The carotid was excised from wild-type Bama pigs,cut into 2.5 cmx 1.0 cm pieces in shuttle shape and subsequently sutured to the abdominal aorta of cynomolgus monkeys.No immunosuppressive agent was administered.General conditions of the recipient monkeys were observed.The morphological changes of the graft artery were assessed by pathological examination at postoperative 1 year.Before and 7,14,28 and 49 d after surgery,the blood samples were collected from the recipient monkeys.The serum levels of IgM and IgG antibodies were quantitatively measured by the red blood cell and peripheral blood mononuclear cell (PBMC) from Bama pigs.The quantity of lymphocytes in the recipient monkeys was detected by routine blood test and flow cytometry.Results All 3 monkeys undergoing transplantation survived well.At postoperative 1 year,the lateral tissues of the vascular wall at the artery graft were seen in dark red color.Hematoxylin-eosin (HE) staining revealed a large quantity of red blood cell and platelet deposition,accompanied with lymphocyte infiltration.Using porcine red blood cell and PBMC as target cells,the serum levels of anti-pig IgM and IgG antibodies peaked at postoperative 28 d,and slightly declined at postoperative 49 d.The quantity of lymphocytes and T cell subset also peaked at postoperative 28 d and began to decrease at postoperative 49 d.Conclusions Artery patch suture is a simple and reliable xenotransplantation model.The recipients can maintain normal physiological state without the use of immunosuppressive agents.The grafts can effectively activate the immune system of the recipients,induce the production of anti-pig antibodies and provoke cellular immune rejection.Therefore,this model can be utilized to monitor the immune rejection throughout the xenotransplantation process.

Sedative effect of administration of different doses of buccal dexmedetomidine in pediatric patients under-going tonsillectomy and/or adenoidectomy / 临床麻醉学杂志

Objective To investigate the sedative effect of different doses of buccal dexmedeto-midine premedication during peri-anesthesia in pediatric patients undergoing tonsillectomy and/or ade-noidectomy.Methods Eighty pediatric patients undergoing tonsillectomy and/or adenoidectomy in department of otorhinolaryngology,54 males and 26 females,aged 4-12 years,ASA Ⅰ or Ⅱ,from June,2014 through May,201 6 were enrolled,n =20 in each group.Children were randomly assigned to receive buccal dexmedetomidine 0 μg/kg (group A),1 μg/kg (group B),2 μg/kg (group C)and 4μg/kg (group D)60 min before transporting to operating room.Sedation score (OAA/S scale)was monitored before and after administering buccal dexmedetomidine.Time of post-operative first spon-taneous respiration,opening eyes,extubation,anxiety score (SAS scale),as well as OAA/S scale, pain intensity (FLACC),and adverse events 60 min after surgery were recorded.Results Compared with group A and group B,markedly superior OAA/S within 60 min after administering buccal dexmedeto-midine in group C and group D were observed (P < 0.05 ).Compared with group A and group B,the OAA/S score 5 min after extubation was lower in group D.FLACC scores within 30 and 60 min after extu-bation in group D were lower than those in group A.Group D showed obviously prolonged time of post-op-erative first spontaneous respiration,opening eyes and extubation compared with the other groups (P <0.05).All the rates of adverse events were similar.Conclusion 2 or 4 μg/kg premedecation of buccal dexmedetomidine 60 min before transporting to operating room can effectively and safely sedate pediatric pa-tients when entered operating room,improve parental separation,mask and sevoflurane acceptance,as well as decrease the stress induced by intubation and post-operative pain.

An integrative bioinformatics study of gene expression profile in prostate cancer / 安徽医科大学学报

Objective To analyze gene expression profile for exploring the function and regulatory network of differ-entially expressed genes in prostate cancer by bioinformatics. Methods The data of gene expression profile in prostate cancer were obtained from GEO database. R software and affy, limma, pheatmap, ggplot2 and other R packages were applied for data mining and bioinformatics analysis. Combined with DAVID and GeneMANIA , dif-ferentially expressed genes and their regulatory networks were annotated. Results These differentially expressed genes with statistical significance were 56 genes, 15 upregulated genes, 41 downregulated genes;these genes were enriched into different subgroups. cav1, slc16a2, cav2, slc16a5, magi2, ptrf, pdlim5, lmod1 and abcc6 were en-riched into the cell membrane component subgroup ofcell component category. cav1, cav2 and ptrf regulated the function of caveolae, they may play an important role in the occurrence and development of prostate cancer. Conclusion Differentially expressed genes between prostate cancer and adjacent tissues assemble a complex regu-latory network. Bioinformatics is a tool for data mining of the regulatory network , which provides ideas and data for the molecular mechanisms in prostate cancer.

Effect of long non-coding RNA HOTAIR on the proliferation and apoptosis of renal cancer cells / 安徽医科大学学报

Objective To investigate the effect of long non-coding RNA (LncRNA) Hox transcript antisense inter-genic RNA ( HOTAIR ) on the cell proliferation and apoptosis of renal cancer cell lines 786-O and ACHN. Methods Small interfere RNA ( siRNA ) that aims to down-regulate HOTAIR expression was transfected into two renal cell lines respectively, and the transfection efficiency was evaluated by qRT-PCR. Then the MTT assay,Ho-chest staining assay and enzyme-linked immunosorbnent assay(ELISA) were used to detect cell proliferation and apoptosis. Results The expression of HOTAIR could be down-regulated effectively by the siRNA (P < 0. 05). Down-regulation of HOTAIR could inhibit the proliferation(P < 0. 05) and increase apoptosis(P < 0. 05) of two re-nal cancer cells. Conclusion HOTAIR plays a role in promoting cell growth of renal cancer.

Preparation and characterization of monoclonal antibody against hTSC29 pep-tides / 中国免疫学杂志

Objective:To prepare monoclonal antibody ( mAb ) against human testis-specific conserved gene ( hTSC29 ) peptides and characterize its immunological and biological features.Methods:According to bioinformatics analysis and prediction of the antigenicity, surface property, hydrophilicity and flexibility of hTSC29, a 18-amino acid residue partial peptide of hTSC29 was synthesized,then immunized the BALB/c mice for preparing antiserum.The mAb against hTSC29 was produced using the routine hybridoma technique.The properties of the mAb against hTSC29 were identified by ELISA, Western blot and immunohistochemistry staining.Results:After cell fusion and subcloning, one hybridoma cell lines secreting specific mAb against hTSC29 protein were obtaind.The Ig subclass of the mAb was IgG2b(κ).ELISA detection showed that the titer of mAbs in cultured was 1∶104.Western blot analysis proved that the mAb could specifically recognize Mr 60 000 protein in human testis total protein.The hTSC29 protein main located at circumference of spermatocyte and spermatid in human testis tissue by immunohistochemistry staining and immunofluorescence assay.Conclusion:One hybridoma cell lines which can secrete specific mAb against hTSC29 protein with high titers and specificity have been established successfully.The mAb will provide efficient tools for functional studies of hTSC29 expressed in spermatogenesis.

Efficacy of plasma procalcitonin in evaluating severity of community-acquired pneumonia in elderly patients / 中华老年医学杂志

Objective To analyze the plasma procalcitonin (PCT) as a predictor of the severity of community acquired pneumonia (CAP) in elderly patients.Methods Totally 90 elderly patients hospitalized with community acquired pneumonia from 2010 to 2011 were analyzed retrospectively for the relation between plasma procalcitonin and severity of pneumonia.All cases were divided into two groups,the severe group (n=36) and the non-severe group (n=54) according to diagnostic criteria.Results The level of plasma PCT was much lower in the severe group (median 2.44 μg/L) than that in the non-severe group (median 0.11 μg/L) (U=335.50,P=0.000).Among all patients,when PCT was lower than 0.5 μg/L,the incidence of non-severe CAP was 76％,however,when PCT was equal or above 2.0 μg/L,the incidence of non-severe CAP was reduced to 9％.In Binary logistic regression analysis,PCT was a risk factor of aged person with severe community acquired pneumonia independent of age and CRUB-65 scores [OR =1.328 (95 ％ confidential interval:1.072,1.645)].PCT had a positive correlation with CRUB-65 scores (U=10.162,P=0.006).In all cases,the patients who improved well had lower PCT value than the remaining (median 0.21 μg/L,17.0μg/L; U=10.000,P=0.000),which also happened in severe cases (median 1.47 μg/L,17.0 μg/L;U=8.000,P=0.000).The area under the receiver operating characteristic curve was 0.872 (95％ confidential interval:0.741,0.914).At a PCT cut-off level of greater than or equal to 2.0 μg/L,the sensitivity and specificity to predict the severity of aged person with CAP was 55.6％ and 98.9％ respectively.Conclusions Plasma PCT may be a good predictor to evaluate the severity of CAP in elderly patients.

Association of MTHFR C677T and MS A2756G polymorphism with semen quality / 中南大学学报(医学版)

OBJECTIVE@#To investigate the association of MTHFR C677T and MS A2756G polymorphism with semen quality in China.@*METHODS@#The experimental group included 75 males with oligospermia, asthenospermia or teratospermia. The control group included 72 fertile males with normal fertility and sperm quality. The differences in the frequency of genetic polymorphism of MTHFR C677T and MS A2756G in the 2 groups were analyzed, and the plasma homocysteine (Hcy) level in both groups was detected.@*RESULTS@#The frequency of MTHFR C677T genotypes (CT, TT and CT+TT) in the abnormal sperm group was higher than that in the control group (P0.05). The Hcy level in abnormal sperm group was higher than that in the control group. In all subjects, the Hcy level of the MTHFR genotypes (CT, TT and CT+TT) was higher than that of the CC genotype, with no difference among the three MS A2756G genotypes.@*CONCLUSION@#CT and TT genotypes of MTHFR C677T are associated with abnormal sperm, which might be part of the pathogenesis of abnormal sperms. T allele may be the risk factor in China. The one mechanism of the association between MTHFR C677T polymorphism and semen quality could be higher Hcy level. MS A2756G polymorphism may not associate with semen quality in China.

Relationship between daytime sleepiness and oxygen desaturation in the male elderly with obstructive sleep apnea/hyponea syndrome / 中华老年医学杂志

Objective To analyze the relationship between daytime sleepiness and oxygen desaturation in the male eldcrly with obstructive sleep apnea /hyponea syndrome (OSAHS).Methods 147 male elderly (aged 60-80 years) with OSAHS in our sleep laboratory from March 2008 to February 2011 were divided into daytime sleepiness and daytime non-sleepiness groups. Epworth sleeping scale (ESS) and polysomnography (PSG) were employed to detect the degree of sleepiness.Oxygen desaturation were evaluated by oxygen desaturation index (ODI),average and minimum blood oxygen,time percentage of 90％ oxygen desaturation (CT90). Resulls There were significant differences between the two groups in body mass index (BMI) (t=4.157),ODI (t=-10.063),average (t=5.800) and minimum (U=916) blood oxygen,CT90(U=887)and apnca/hypopneaindex (AHI) (t =-11.361) (all P＜0.001).ESS scores were related with desaturation parameters of ODI,average and minimum blood oxygen and CT90 (r=0.683,-0.450,-0.583,0.507,all P＜0.001).ODI was correlated with average and minimum blood oxygen and CT00 (r=-0.628,-0.763,0.689,all P＜0.001).Among parameters of oxygen desaturation,only ODI had influence on ESS scores (odds ratio=1.11,95％CI:1.07 1.15),The sensitivity and specificity predicting daytime sleepiness at ODI ≥23 times/h were about 79.2％ and 84.3％,respectively.Conclusions Daytime sleepiness is associated with oxygen desaturaion and may be predicted by ODI through which average and minimum blood oxygen and CT00 are related with daytime sleepiness in the male elderly with OSAHS.

Relationship between obstructive sleep apnea-hypopnea syndrome and metabolic syndrome in obese middle-aged and older men / 中华老年医学杂志

Objective To explore the relationship between obstructive sleep apnea-hypopnea syndrome (OSAHS) and metabolic syndrome (MS) in obese middle-aged and older men. Methods We selectively recruited 154 obese middle-aged and older men matched for body mass index (BMI) and age. The polysomnography was performed for diagnosing OSAHS and for discriminating disease severity. The BMI, waist circumference, blood pressure, plasma glucose and lipid profiles were measured and analyzed in all subjects. Appropriate statistical methods were used to compare the components of MS in each group. Logistic regression was taken to elucidate the relationship between OSAHS and MS. Results Compared to control group, severe OSAHS group had significantly lower high density lipoprotein cholesterol level [( 1.03 ± 0.29 ) mmol/L vs. ( 1.31 ± 0. 38) mmol/L,P＜0. 05] and higher fasting glucose [(6.61±1.76) mmol/L vs. (5.47±0.64) mmol/L, P＜0. 05]as well as higher systolic blood pressure [( 133 ± 13) mm Hg vs. ( 125 ± 12) mm Hg, P＜0. 05] and diastolic blood pressure [(99±10) mm Hg vs. (80±5) mm Hg, P＜0. 05]. The prevalence of MS was significantly higher in OSAHS group than in control group (mild OSAHS group: 25.7%,moderate OSAHS group: 46. 5%, severe OSAHS group: 84.4%, control group: 16. 1 %, all P＜0. 01). OSAHS was independently associated with an increased prevalence of MS Odds ratio, 6.16).Conclusions OSAHS is independently associated with MS in obese middle-aged and older men.

Expression of Tpap gene in mouse testis / 中国组织工程研究

BACKGROUND: These serial processes for forming male gametes are basically controlled by the programmed expression of a number of stage-specific genes. However, many aspects of the mechanisms of spermatogenesis have remained elusive because of a lack of suitable in vitro or in vivo models.OBJECTIVE: To screen genes involved in spermatogenesis, and to analyze its expression characteristics. METHODS: Testes cDNA samples from Balb/C mice of different postnatal days (4,9,18,35, 54 days and 6 months, respectively) were hybridized with mouse whole genome Affymetrix chip to screen the testis-ralated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. RT-PCR was used here to identify the expression of the selected genes in mice testis.RESULTS AND CONCLUSION: The Affymetrix chip probe of mouse Tpap was graduated higher expression with developmental stages of mouse testis. The scaling hybridization signal intensities of the tested testis on days 4, 9,18, 35, 54, and 6 months of postnatal were 4.4 (Absent expression, A), 12.9 (A), 262.4 (Present expression, P), 1136.7 (P), 1617.5 (P) and 1128 (P),respectively. These results indicated that the expression of mouse Tpap wasn't detected on days 4 and 9, but was detected on days 18, 35, 54, and 6 months of mouse testis in our Affymetrix chip analysis. By combination with the RT-PCR analysis of mouse Tpap, we observed mouse Tpap began to express at the age of day 18 in mouse. Tpap is an age-dependent gene in mouse testis.The expression of Tpap corresponds to the appearance of spermatids of mice and indicates that Tpap may have an important role in male mammalian spermatogenesis.

Expression profile of a novel germ cell-specific gene, TSCPA, in mice and human / 华中科技大学学报（医学）（英德文版）

In order to identify novel genes involved in spermatogenesis, testis cDNA samples from Balb/C mice of different postnatal days were hybridized with the whole mouse genome Affymetrix chip to screen the testis-specific genes. The characteristics of the selected genes were analyzed by RT-PCR as well as other bioinformatic tools. A novel differentially expressed testis-specific gene (GenBank Accession No: NM_029042) in the developmental stages of testes was identified, and named TSCPA. Cellular mapping prediction of TSCPA indicated that its protein was probably expressed in nuclei, and one putative domain (aa 332-377) was anchoring domain of cAMP-dependent type II PK. The result of subcellular localization of GFP-TSCPA fusion protein in Cos-7 cells showed that TSCPA protein was expressed in nuclei. RT-PCR analysis revealed that TSCPA was expressed specifically in mouse and human testis. TSCPA gene was expressed weakly in 21-day-old mouse testis and the expression was increased gradually from 38th day to 6th month of mouse testes. No expression of hTSCPA was found in cryptorchidism and Sertoli-cell-only syndrome patients. It was concluded that the expression profile of TSCPA in human and mice indicated that TSCPA might play an important role in spermatogenesis.

Development of early human fetal testes after xenografting into mice / 解剖学报

Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of human spermatogenetic mechanism.

Preliminary study on the development of germ cells from human fetal testicular tissues xenografted into the mouse / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the development of xenografted primitive human germ cells by using fetal testicular tissues as donor tissues and an immunodeficient mouse as the recipient.</p><p><b>METHODS</b>Testicular tissue fragments of a 26-week fetus were grafted under the back skin of a castrated immunodeficient mouse. Grafts were taken out after 135 days and processed for morphological and histological analyses.</p><p><b>RESULTS</b>The mass of grafts grew from about 1 mm in diameter and 5 mg in wet weight to about 3 mm and more than 20 mg 135 days after grafting. Histological observations showed a significant expansion of seminiferous tubules after grafting (80 +/- 25 microm in diameter) in comparison with seminiferous cords at the time of grafting (60 +/- 15 microm in diameter). The seminiferous cords developed into seminiferous tubules with the epithelial border and lumen. After 135 days of grafting, most of the dispersedly distributed primitive Sertoli cells and germ cells migrated to the basal part of seminiferous epithelium, located on the basement membrane and few of germ cells differentiated into spermatogonia.</p><p><b>CONCLUSION</b>Human fetal testicular tissues could survive and continuously develop after being xenograft into castrated immunodeficient mice.</p>

Expression of amphiregulin in human endometrium during the menstrual cycle / 北京大学学报(医学版)

Objective:To investigate the expression of amphiregulin in human endometrium during the menstrual cycle.Methods: Endometrial tissues were collected from the patients undergoing hysterectomy or endometrial biopsy.Real-time RT-PCR,in situ hybridization and immunohistochemistry were used to detect the expression characteristics of amphiregulin in human endometrium in proliferative and secretory phases.Results: Real-time RT-PCR showed the expression of amphiregulin mRNA in secretory phase was 32 times that in proliferative phase.The results from in situ hybridization and immunohistochemistry showed that amphiregulin was located in the cytoplasm and mainly expressed in the gland of endometrium.The expressions of amphiregulin mRNA in proliferative and secretory phases were 0.54?0.22 and 2.96?0.47(P

Hormone regulation of the expression of vascular endothelial growth factor and its receptors in mouse uterus / 北京大学学报(医学版)

Objective: To investigate the effects of estrogen and progesterone on the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR) in mouse uterus. Methods: 3-week-old immature female mice were randomly divided into 7 groups and treated with corn oil, estradiol (E2) of 1.5, 3.0, 10, 25 ng, progesterone (P) of 100 ?g and (E_2 10 ng + P 100 ?g)/mouse, respectively. After the treatment for 48 h, mouse uterus was collected to isolate total RNA. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA isoforms of VEGF and its receptors in mouse uterus. Results: Compared with control, both E_2 and P significantly increased the expression of VEGF164 and VEGF120 mRNA in mouse uterus. The expression of VEGFR2 mRNA, not VEGF1 mRNA, was decreased by E_2 treatment in a dose-independent manner. Conclusion: Both estradiol and progesterone up-regulated the expression of VEGF, but estradiol down-regulated the expression of VEGFR2 in mouse uterus.