Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development / 中华儿科杂志

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

Micro-dissection testicular sperm extraction for non-obstructive azoospermia patients with the history of secondary testicular injury / 中华男科学杂志

<p><b>Objective</b>To investigate the value of micro- dissection testicular sperm extraction (micro-TESE) in the treatment of non-obstructive azoospermia (NOA) in patients with the history of secondary testicular injury.</p><p><b>METHODS</b>Totally, 121 NOA patients with the history of secondary testicular injury underwent micro-TESE in our hospital from September 2014 to December 2017. We analyzed the correlation of the sperm retrieval rate with the causes of testicular injury and compared the outcomes of the ICSI cycles with the sperm retrieved from the NOA males by micro-TESE (the micro-TESE group) and those with the sperm ejaculated from severe oligospermia patients (sperm concentration <1×10⁶/ml, the ejaculate group). Comparisons were also made between the two groups in the female age, two-pronucleus (2PN) fertilization rate, transferrable embryos on day 3 (D3), D3 high- quality embryos, D14 blood HCG positive rate, embryo implantation rate, and clinical pregnancy rate.</p><p><b>RESULTS</b>Testicular sperm were successfully retrieved by micro-TESE in 86.0% of the patients (104/121), of whom 98.4% had the history of orchitis, 75.5% had been treated surgically for cryptorchidism, and 63.6% had received chemo- or radiotherapy. No statistically significant differences were observed between the micro-TESE and ejaculate groups in the 2PN fertilization rate (59.4% vs 69.3%, P > 0.05), D14 blood HCG positive rate (44.6% vs 57.9%, P > 0.05), embryo implantation rate (31.8 %% vs 32.6%, P > 0.05) and clinical pregnancy rate (41.5% vs 48.7%, P > 0.05). However, the rate D3 transferrable embryos was significantly lower in the micro-TESE than in the ejaculate group (40.5% vs 52.2%,P < 0.05), and so was that of D3 high-quality embryos (32.5% vs 42.1%, P < 0.05).</p><p><b>CONCLUSIONS</b>Micro-TESE can be applied as the first choice for NOA patients with the history of secondary testicular injury, but more effective strategies are to be explored for the improvement of ICSI outcomes with the sperm retrieved by micro- TESE.</p>

Micro-dissection testicular sperm extraction for patients with non-obstructive azoospermia: A report of 196 cases / 中华男科学杂志

Objective@#To investigate the effect of micro-dissection testicular sperm extraction (microTESE) for patients with non-obstructive azoospermia (NOA) and the indications of the strategy.@*METHODS@#This retrospective study included 196 cases of NOA undergoing microTESE in our center from September 2014 to March 2017. We recorded the sperm retrieval rate (SRR) and analyzed its correlation with the patients' age, testis volume, level of blood follicle-stimulating hormone (FSH), and etiological factors.@*RESULTS@#Testicular sperm were successfully retrieved from 87 (44.4%) of the patients. No significant correlation was found between the SRR and the patients' age, testis volume, or blood FSH level (P >0.05). As regards etiological factors, the SRR was 100% (29/29) in the patients with orchitis, 66.7% (16/24) in those surgically treated for cryptorchidism, 55.6% (10/18) in those with other secondary testis lesions, 60.0% (3/5) in those with AZFc deletion, 40.9% (9/22) in those with severe idiopathic testicular atrophy, 21.4% (12/56) in those with idiopathic NOA, 20.5% (8/39) in those with Klinefelter's syndrome, and 0% (0/3) in those with other abnormal karyotypes.@*CONCLUSIONS@#MicroTESE is an effective strategy for sperm retrieval in NOA patients, and the SRR is correlated with etiological factors but not with the FSH level or testis volume of the patients.

Expressions of hypoxia inducible factor 1 alpha and P53 in the epididymis of varicocele rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of epididymal hypofunction of rats with varicocele (VC) by observing the changes in the epididymal index, motility of epididymal sperm, expressions of hypoxia-inducible factor 1 alpha (HIF-1 alpha) and the tumor suppressor protein p53, and epididymal epithelial cells.</p><p><b>METHODS</b>Ninety SD rats were equally randomized to a VC model (A), a sham operation (B), and a normal control group (C). At 49 days after surgery, all the rats were executed after weighing. Then the volume of the left epididymis was obtained, the epididymal sperm motility was detected by computer-assisted sperm analysis (CASA), the expressions of HIF-1 alpha and p53 in the epididymal tissue were determined by Western-blot, and the epididymal epithelial cells were observed by HE staining.</p><p><b>RESULTS</b>VC models were successfully established in 27 of the rats. One-way ANOVA test showed no statistically significant differences in the epididymis index among groups A ([40.53 +/- 1.76] x 10 (-5)) , B ([43.31 1.58] x 10( -5)) , and C ( [44. 10 +/- 2.62] x 10 -5) (P > 0.05). Sperm motility and the percentage of progressively motile sperm were significantly lower in group A ([71.86 +/- 5.07]% and [42. 26 +/-4.45]%) than in B ([78.51 4.50]% and [49.08 +/-4. 19]% ) and C ( [79.24 +/- 2.70] % and [52. 23+/- 2. 23] % ) (both P <0.05) , while the expressions of HTF-1 a and p53 were remarkably higher in A (1.74 +/- 0. 16 and 1.71 +/- 0. 11) than in B (0.32 +/- 0. 08 and 0.56 +/- 0.13) and C (0.12 +/- 0. 03 and 0.25 +/-0.06) (both P < 0.05). The epididymal epithelial cells in group A were obviously decreased in number and arranged in loose and disorderly patterns as compared with those in B and C.</p><p><b>CONCLUSION</b>Varicocele can cause hypoxia in the epididymal tissue, which in turn may lead to epididymal hypofunction.</p>

Gene-gene interaction of GJB2, SOD2, and CAT on occupational noise-induced hearing loss in Chinese Han population / 生物医学与环境科学（英文）

The effects of genetic factors on the noise-induced hearing loss (NIHL) are still unclear. In the present study, eight single-nucleotide polymorphisms (SNPs) included rs1227049 and rs3802711 (CDH23), rs1695 (GSTP1), rs137852540 (GJB2), rs2289274 (PMCA2), rs4880 (SOD2), rs7943316, and rs769214 within CAT that might associated with NIHL were further validated in Chinese workers. The results showed that the carriers of the T allele (AT+TT) of rs7943316 and A allele (GA+AA) of rs769214, were significantly associated with an increased risk of NIHL compared to those with AA genotype (P<0.05) and GG genotype (P<0.05). Moreover, a significant three-locus model (P=0.0107) involving rs2016520, rs9794, and rs1805192 were observed that might associated with NIHL, with 53.95% of testing accuracy. Thus, our present study provided the evidence that GJB2, SOD2, and CAT genes might account for the NIHL development in independently and/or in an interactive manner.

Roles of peroxisome proliferator-activated receptors polymorphisms, haplotypes, levels on C-reactive protein and their interactions with abnormal body weight / 中华流行病学杂志

Objective To explore the roles of peroxisome proliferator-activated receptors (PPARs) on the levels of serum C-reactive protein (CRP) and the interactions of PPARs haplotypes with abnormal body weight.Methods Subjects (n=644) were randomly selected from the cohort ‘Prevention of Multiple metabolic disorders and Metabolic syndrome in Jiangsu province (PMMJS)'Variance test,t test and lineal regression were used to analyze the associations between PPARs polymorphisms and the levels of CRP.The association between PPARs haplotypes and serum CRP levels as well as the interaction of PPARs haplotypes with abnormal body weight were analyzed,under the SNPStats software.Results After adjusting for sex,age,blood pressure,cigarette smoking,alcohol drinking and so on,data showed that both rs1800206 and rs9794 were associated with the changes along with the levels of CRP (P＜0.05).After adjusting for the same factors,haplotypes of AVG and CVG in PPARα,CG in PPARδ appeared to be associated with the increase (P＜0.05) while haplotypes of CC in PPARδ,CPCAC in PPARγ were associated with the decrease of CRP levels (P＜0.05).Results from the Interaction analysis also noted that the interactions did exist between abnormal body weight and both AVG,CVG in PPARαt,and CG in PPARδ.Conclusion PPARs polymorphisms and haplotypes were associated with CRP.Interaction between PPAR α/δand abnormal body weight might contribute to the levels of CRP.

Clinical characteristics of pediatric victims in the Lushan and Wenchuan earthquakes and experience of medical rescue / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To get a more comprehensive understanding of the clinical characteristics of pediatric victims in earthquake and to summarize the experience of medical rescue.</p><p><b>METHODS</b>The clinical information was collected from the pediatric victims who were admitted to West China Hospital, Sichuan University following the Lushan earthquake in 2013 and Wenchuan earthquake in 2008. The clinical data were compared between the pediatric victims in the two earthquakes.</p><p><b>RESULTS</b>Thirty-four children under 14 years of age, who were injured in the Lushan earthquake, were admitted to the West China Hospital before April 30, 2013. Compared with the data in the Wenchuan earthquake, the mean age of the pediatric victims in the Lushan earthquake was significantly lower (P<0.01), and the mean time from earthquake to hospitalization was significantly shorter (P<0.01). In the Lushan earthquake, 67.6% of the injured children had variable limb fractures; traumatic brain injury was found in 29.4% of hospitalized children, versus 9.5% in the Wenchuan earthquake (P<0.05). Among the 34 children, no amputation and death occurred, and all the 13 severe cases started to recover.</p><p><b>CONCLUSIONS</b>There were higher proportions of severely injured children and children with traumatic brain injury in the Lushan earthquake than in the Wenchuan earthquake. But these cases recovered well, which was possibly due to timely on-site rescue and transfer and multi-sector, multi-institution, and multidisciplinary cooperation.</p>

Induced differentiation of human adipose-derived stem cells into Leydig cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the feasibility of inducing the differentiation of human adipose tissue-derived stem cells (ADSCs) into Leydig cells in vitro.</p><p><b>METHODS</b>We isolated ADSCs by digestion with Collagenase I from the subcutaneous adipose tissue, cultured them in the DMEM/F12 medium with 10% fetal bovine serum, and detected the expression of vimentin by immunohistochemistry. We exposed the ADSCs to different concentrations of human chorionic gonadotropin (HCG) for different times, determined the expression of StAR mRNA by real-time PCR, and measured the HCG-induced proliferation of ADSCs by MTT. After a week of induction by HCG and DMSO, we conducted 3beta-HSD immunohistochemistry, and detected the testosterone level in the supernatant and lysis of the cells by radioimmunoassay.</p><p><b>RESULTS</b>The ADSCs grew well with a positive expression of vimentin. The expression of the StAR gene was positively correlated with the increased concentration of HCG, reaching the peak at HCG 10 U/ml in 1 week culture. The proliferation of ADSCs was significantly increased by HCG induction. A positive expression of 3beta-HSD was observed after 1 week induction with HCG 10 U/ml and DMSO 3.2 x 10(-6)mol/L.</p><p><b>CONCLUSION</b>HCG enhances the expression of the StAR gene and the proliferation of ADSCs. Induced by HCG 10 U/ml and DMSO 3.2 x 10(-6) mol/L, ADSCs tend to differentiate into Leydig cells.</p>

Identification, structure and function of a novel tetraspaninhomologue from Spirometra erinaceieuropaei

OBJECTIVE@#To identify a full length c DNA sequence of a novel tetraspanin (TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.@*METHODS@#Using the NCBI, EMBI, Expasy and other online sites, the open reading frame (ORF), conserved domain, physical and chemical parameters, signal peptide, transmembrane domain, epitope, topological structures of the protein sequences were predicted. And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.@*RESULTS@#The target sequence was 1132 bp length with a 681 bpbiggest ORF encoding 226 amino acids protein with typical TSP conserved domain. It was confirmed as full length c DNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number: JF728872). The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da, respectively. Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni, it showed similarity of 59% and 59%, respectively. SeTSP16 contained four transmembrane domains (TM1-4), intracellular N and C-termini, one short small extracellular loop and one large extracellular loop. Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.@*CONCLUSIONS@#The full length c DNA sequences of SeTSP16 are identified. It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.

In utero exposure to dichlorvos induces apoptosis of Leydig cells in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the influence of the organophosphate insecticide dichlorvos on the apoptosis of Leydig cells in the male offspring of the SD rats exposed to dichlorvos, and to investigate the role of the changes of Leydig cells in genitourinary malformation.</p><p><b>METHODS</b>Twenty-one pregnant SD rats were divided into a corn oil control group and 6 dichlorvos groups, the former given by gavage 1.0 ml corn oil daily, and the latter dichlorvos at the dose of 1, 4, 8, 16, 20 and 24 mg/kg daily from the 12th to 17th day of conception. After birth, 5 male neonates were randomly selected from each of the control and dichlorvos groups, and their testes were harvested to be analyzed by HE staining, immunohistochemistry with anti-caspase-3 antibodies and DAPI fluorescent staining. At 90 days after birth, another 5 of the male offspring were taken from each group and their testes were collected for the same analyses.</p><p><b>RESULTS</b>Statistically significant differences were found in the number of both the caspase-3 positive and DAPI labeled Leydig cells in the testes of the rat offspring between the corn oil and the 4, 8, 16, 20 and 24 mg/kg dichlorvos groups (P < 0.05), but not between the control and the 1 mg/kg dichlorvos groups (P > 0.05). The apoptosis of Leydig cells was increased in the male offspring of the dichlorvos-exposed SD rats in a dose-dependent manner.</p><p><b>CONCLUSION</b>Exposure of pregnant rats to dichlorvos can increase the apoptosis of Leydig cells in the male offspring, which, in turn, may reduce the number of Leydig cells, interfere with the testis function during the embryonic period, and damage the development of the genitourinary system.</p>

Effect of fluvastatin on vascular endothelial growth factor in rats with osteoporosis in process of fracture healing / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To explore the effect of fluvastatin on vascular endothelial growth factor (VEGF) in rats with osteoporosis in the process of fracture healing.</p><p><b>METHODS</b>Fractures at the intermediate piece of the femur were made on 72 Sprague Dawley (SD) rats (weighing initially 290-340 g and aged 6 months) with osteoporosis after ovariectomy for three months, then these rats were divided randomly into the medication administration group (the experimental group) and the control group, 36 rats each. In the experimental group, the rats received fluvastatin lavage (10 mg/kg per day) since the next day of operation lasting for 6 weeks, and the rats in the control group received placebo. Then the expression of VEGF and VEGF mRNA in bony callus of the two groups was measured respectively with immunohistochemistry and in situ hybridization on days of 3rd, 7th, 14th, 21st, 28th, and 42nd, and image analysis was made with real-color image analysis machine.</p><p><b>RESULTS</b>No difference was found in the cellular localization of VEGF and VEGF mRNA gene expression between the experimental group and the control group in process of fracture healing and their expression modes were almost similar. On the 14th day postoperatively, the positive extent of positive cells in the experimental group was higher than that of the control group (P < 0.05).</p><p><b>CONCLUSION</b>Fluvastatin can promote the VEGF level in rats with osteoporosis in process of fracture healing.</p>

Pathological changes in the testes of the rats with hypospadia induced by dichlorvos / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the mechanism of dichlorvos leading to hypospadia of rats.</p><p><b>METHODS</b>From the 12th to the 17th day of conception, 20 pregnant female rats (the experiment group) were given 10 mg/(kg x d) dichlorvos, while another 10 (the control group) administered 1.5 ml 0.9% NaCl/day. Out of 88 male newborns of the 20 experimental mother rats, 22 had hypospadia, while out of the 33 male newborns of the 10 controls, none had the problem. Five hypospadia newborns from the experiment group and another 5 normal ones from the control group were raised to sexual maturity, and then their testes were excised and embedded in paraffin, and the tissue sections were analyzed by regular HE staining and SP immunohistochemical staining with Calretinin.</p><p><b>RESULTS</b>HE staining showed that the number of Leydig cells in the testis tissues of the hypospadia rats decreased significantly compared with the normal ones, but no change was observed either in the number or in the morphology of the seminiferous tubules. Moreover, the Calretinin positive Leydig cells were reduced dramatically in the testes of the hypospadia rats.</p><p><b>CONCLUSION</b>Pregnant female rats, when exposed to dichlorvos, may cause reduction of testis Leydig cells in their male offsprings. Thus the probable mechanism of rat hypospadia induced by dichlorvos may lie in the decrease of the testosterone level caused by damage to Leydig cells from dichlorvos toxicity.</p>

A prospective study of the anterior cruciate ligament reconstruction: allograft versus autograft / 中华外科杂志

<p><b>OBJECTIVE</b>To compare the clinical outcomes and differences of arthroscopic anterior cruciate ligament (ACL) reconstruction by using allografts and autografts.</p><p><b>METHODS</b>Fifty-three patients with grade III ACL injuries were divided randomly into 2 groups. Twenty-five cases of group A with autografts of patellar bone-tendon-bone (B-PT-B;n = 15) and 4 strands of semitendinosus/gracilis (n = 10); 28 patients of group B with allografts of B-PT-B (n = 18), 4 strands of semitendinosus/2 strands of gracilis tendons (n = 6) and double tibialis posterior (n = 4) as well as Achilles bone-tendon (n = 2). All procedures were performed endoscopically by some surgeons. All patients were followed up for 12 to 31 months, average 19 months.</p><p><b>RESULTS</b>There were statistically significant differences revealed in physical examination. IKDC and Lysholm-Tegner Score as well as KT-2000 testing was used preoperation and postoperation for both group, but there were no significant differences between the two groups except that group B had less time in operation and longer time in postoperative fever comparing with group A. (2-sample t test, P < 0.05). The KT-2000 side to side difference less than 3 mm were 88% and 86% and more than 5 mm were 4% and 7.1% respectively. The infection rates were 0% and 3.5% for the two groups.</p><p><b>CONCLUSION</b>The clinical outcomes of the allografts and autografts in ACL reconstruction is almost the same. Allografts are a reasonably alternative choice for ACL reconstruction in patients over middle ages and for multiple ligament surgery as well as revision procedures.</p>