High expression of follicle stimulating hormone receptor in testicular tissue of idiopathic azoospermic patients with severe spermatogenic defects / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Follicle stimulating hormone is necessary for normal reproduction in men. The biochemical actions of follicle stimulating hormone result from binding to the follicle stimulating hormone receptor in the plasma membrane of Sertoli cells. Here, we investigated the expression of the follicle stimulating hormone receptor in different testicular histological phenotypes of patients with idiopathic azoospermia.</p><p><b>METHODS</b>Fifty-seven cases of idiopathic azoospermia were classified into three groups according to the results of testicular biopsy: patients with hypospermatogenesis, patients with maturation arrest, and patients with Sertoli cell-only syndrome. Thirteen azoospermic patients identified by testicular biopsy as being capable of completing spermatogenesis acted as the control group. Immunohistochemistry and real-time quantitative reverse-transcriptase polymerase chain reaction were performed in each case, and the serum hormone level was also measured in all patients.</p><p><b>RESULTS</b>The serum follicle stimulating hormone level in patients with Sertoli cell-only syndrome was significantly higher than in patients with hypospermatogenesis, maturation arrest, and complete spermatogenesis (P < 0.01). The serum follicle stimulating hormone level in patients with maturation arrest was significantly higher than in patients with hypospermatogenesis and complete spermatogenesis (P < 0.05). There was no difference in serum follicle stimulating hormone levels in patients with hypospermatogenesis and complete spermatogenesis. The follicle stimulating hormone receptor expression level of testicular samples with Sertoli cell-only syndrome was significantly higher than in those with hypospermatogenesis, maturation arrest, and complete spermatogenesis (P < 0.05), but no significant difference was observed among hypospermatogenesis, maturation arrest, and complete spermatogenesis testicular samples.</p><p><b>CONCLUSIONS</b>Different serum follicle stimulating hormone levels and follicle stimulating hormone receptor expression were found in the different testicular histology phenotypes in azoospermic patients. Differential follicle stimulating hormone receptor expression in testicular tissue of patients with idiopathic azoospermia may be associated with the degree of spermatogenesis.</p>

Morphology and cell proliferation evaluation of follicles from cryopreserved human ovarian tissue by vitrification / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the morphology and proliferation of follicles from cryopreserved human ovarian tissue by vitrification.</p><p><b>METHODS</b>Ovarian biopsy specimens were taken from 12 patients. The specimens were randomly distributed into fresh group (Group A) and vitrification group (Group B). Histological examination and ultrastructural observation were performed after cryopreservation. Both were embedded in paraffin block and proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical staining.</p><p><b>RESULTS</b>The proportions of primordial and primary follicles from Group A and Group B were 86.4%, 13.6% and 84.5%, 15.5%, respectively (P>0.05). There was no significant difference in proportions of morphologically normal primordial follicles between Group A and Group B (P>0.05); but the proportion of morphologically abnormal primary follicles was significantly higher in Group B than that in Group A (P<0.05). The ultrastructural studies showed that in histologically normal primordial follicles, there was no difference between Group A and Group B, while there were a few abnormalities of primary follicles in Group B. Granulosa cells and oocytes of primordial and primary follicles and stromal cells were positive for PCNA staining both in fresh and cryopreserved ovarian tissues; there were no differences between two groups.</p><p><b>CONCLUSION</b>Vitrification is a favorable method in human ovarian cryopreservation.</p>

Preimplantation genetic diagnosis of translocation / 中华妇产科杂志

Objectives To observe the genetic characteristics of chromosomes and the rates of implantation and pregnancy in couples of translocation carriers who undergo preimplantation genetic diagnosis (PGD) and to evaluate the significance of PGD in the treatment of translocation carriers. Methods Fluorescence in situ hybridization (FISH) was performed to analyze the embryos of 12 carriers of reciprocal translocation and 22 carriers of Robertsonian translocation. The results of diagnosis and the implantation and pregnancy rates were analyzed. Results A total of 253 embryos from 36 couples were retrieved and FISH was applied for the examination. The characteristics of chromosomes were diagnosed in 225 embryos and the rate of successful PGD was 88.9%. Fifty-eight embryos were found to have normal chromosome or balanced translocation and were transferred into the uterus. The rate of implantation was 36% (5/14) and 14% (6/44) and the rate of pregnancy was 4/9 and 26% (5/19) for carriers of Robertsonian translocation and reciprocal translocation, respectively. Conclusions The FISH-based PGD is effective in the diagnosis of Robertsonian translocation and reciprocal translocation of embryos. It provides the possibility of a high rate of implantation and pregnancy, and avoids recurrent abortion and unwilling termination of pregnancy.

Glutathione S-transferase M1 and T1 genotypes and endometriosis risk: a case-controlled study / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the correlation between glutathione S-transferase (GST) M1 and T1 genotypes and endometriosis risk (EM).</p><p><b>METHODS</b>Polymerase chain reaction (PCR) technique was used to detect the presence or absence of the GSTM1 and GSTT1 genes in genomic DNA isolated from the blood samples of 68 Han Chinese women with endometriosis and 28 without endometriosis.</p><p><b>RESULTS</b>The frequencies of GSTM1 and GSTT1 null genotypes in women with endometriosis were 0.721 (49/68) and 0.779 (53/68), respectively, and in women without endometriosis were 0.429 (12/28) and 0.321 (9/28), respectively. There was a significant difference with regard to the frequencies of GSTM1 and GSTT1 null genotypes between the women with and without endometriosis (P < 0.01). Furthermore, the frequencies of GSTM1 and GSTT1 null genotypes were significantly higher in the patients with stage III and IV endometriosis [0.731 (38/52) and 0.788 (41/52), respectively] than in women without endometriosis (P < 0.01), and the frequency of GSTT1 null genotype was statistically higher in patients with stage I and II endometriosis [0.75 (12/16)] than in the women without endometriosis (P < 0.01). No correlation between GSTM1 and GSTT1 null genotypes and age, induced abortion or dysmenorrhea was detected in this study (P > 0.05).</p><p><b>CONCLUSION</b>GSTM1 and GSTT1 null genotypes may be risk factors for the development of endometriosis.</p>

Polymorphism in insulin receptor gene exon 17 in women with polycystic ovary syndrome / 中华妇产科杂志

Objective To investigate insulin receptor (INSR) genotype exon 17 frequencies in women with polycystic ovary syndrome(PCOS) and to elucidate its role in the pathogenesis of PCOS. Methods The study involved 33 women with PCOS and 28 healthy control women who were genotyped for polymorphism of INSR gene exon 17 by single strand conformation polymorphism (SSCP) analysis. Body mass index (BMI), insulin sensitive index (ISI), the expression of INSR beta subunit, and serum concentration of luteinizing hormone(LH), total testosterone between the genotypes were compared. Results (1) The T -to- C mutation was observed in the INSR gene exon 17 (1008 bp). The frequency of the C/C genotype was significantly higher in patients (39%) than in the controls (11%) (P

Cloning of a new human cytochrome P450 2A6 cDNA / 中国病理生理杂志

AIM:To clon human cytochrome P450 2A6 cDNA. METHODS:Using reverse transcription polymerase chain reaction(RT-PCR) and DNA recombinant technique, a full-length cDNA encoding cytochrome P450 2A6( CYP2A6 ) from human liver was cloned into pBluescript vector. The cDNA segment was identified by DNA sequencing.RESULTS: Comparing with the CYP2A6 sequence, the cloned CYP2A6 cDNA had two different bases, codon 8 CTG(Leu)→TTG(Leu), codon 479 GGC(Gly)→GTC(Val) and was quite different in their 3′end noncoding region. Comparing with CYP2A7 seqence reported by Fernandez-Salguero,the cloned CYP2A6 cDNA had some different in 5′ end coding sequence and several differencey in the 3′ end coding and noncoding sequence, but both codon 479 were GTC(Val).Comparing with the CYP2A7 seqence reported by Yamano,the cloned CYP2A6 cDNA had some difference in the coding sequence but the 3′ end no-coding area was the same. CONCLUSION: The cloned cDNA was a new cDNA of CYP2A6 which may be transcripted from a new allele of CYP2A6.

Nifedipine induced reversal of multidrug resistance can be deteriorated by S9 mix prepared from the transgenic cell line CHL-3A4 / 中国病理生理杂志

AIM:To demonstate the expression of human cytochrome P450 3A4 isozyme in the transgenic cell line CHL - 3A4 established in this laboratory. METHODS:Nifedipine (NIF) can induce the reversal of the brug resistance for adriamycin of multidrug resistant(MDR) cell K562r. The NIF is the specific substlate for CYP3A4. To determine the NIF oxidase activity of the transgenic CHL - 3A4 cells by comparing the biological effect of NIF, which preinculbated with or without CHL - 3A4 S9 mix. RESULTS: When the multidrug resistance cell K562r was cultured in medium with NIF (12. 5 ?g? L- 1 ), a calcium channel blocker and specific substrate for CYP3A, it's IC50 for adri- amycin declined from(6.47?0.60) mg ? L- 1 to (0. 89?0. 15) mg? L-1. When cells were cultured in NIF(exactly the same concentration ) pretreated with CHL - 3A4 S9 mix,no reversal of MDR was observed [IC50 for adriamycin is (6.10?0.50) mg? L-1 ] while if NIF was pretreated with CHL S9 or inactivated CHL - 3A4 S9 mix, its biological effect was not deteriorated [IC50 for adriamycin is (0. 32?0.90) and (0.32?0.04) mg?L-1, P

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3 / 中国病理生理杂志

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-?-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78 6 kD on SDS-PAGE analysis,but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.