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.

Primary gastric Burkitt's lymphoma: a clinicopathological analysis of four cases / 临床与实验病理学杂志

Purpose To investigate the clinicopathological features,diagnosis,treatment and prognosis of primary gastric Burkitt's lymphoma. Methods The clinical and pathological features were retrospectively analyzed in 4 case of primary gastric Burkitt's lymphoma. The endoscopic features,immunophenotype and molecular pathology were also analyzed and discussed with review of the literatures. Results Primary gastric Burkitt' s lymphoma mainly occurred in children,with an average age of 8 years (3～15 years) . Endoscopic findings were tuberous protuberance with surface ulcers (3 cases) . The diffuse distribution of atypical lymphoid cells with"starry sky"phenomenon in gastric mucosa was observed under microscope,the cells were of medium size and single shape,the nucleus was round or oval and the shape was uniform,the chromatin was thicker,and there were several small nucleoli,the cytoplasm was rare and slightly basophilic,and the nuclear mitosis was more frequently noted. Immunophenotyping showed that both CD20 and CD79a were all strongly positive,and the Ki-67 proliferative index was higher than 95％. The positive intensity of CD10 and BCL-6 varied from one to another. MUM1 and CD3 were all negative. One case was positive for EBER (in situ hybridization) . Meanwhile, all of these cases showed MYC gene breakage,one of them was detected by IGH/BCL-2 fusion gene but no fusion was found. Conclusion Primary gastric Burkitt's lymphoma is rare. Its endoscopic features,histomorphology,immunophenotype and molecular pathology are relatively typical and it is mainly distinguished from high-grade B cell lymphoma,lymphoblastic lymphoma and myeloid sarcoma. Burkitt's lymphoma is a highly malignant tumor,but the current treatment is relatively mature. Early diagnosis and early treatment can significantly improve the prognosis of these patients.

Expression of c-MET, EGFR and HER-2 of human gastric adenocarcinoma / 临床与实验病理学杂志

Purpose To investigate the expression status and their clinical significances of c-MET, epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 (HER-2) in gastric adenocarcinoma (GC). Methods Tissue samples from 442 cases of GC in patients who accepted D2/D3 radical gastrectomy with R0 resection were stained by immunohistochemistry against c-MET, EGFR and HER-2.Results Over expression of c-MET, EGFR and HER-2 was identified in 195/442 (44.1％), 47/442 (10.6％) and 152/442 (34.4％) GC patients, respectively. Over expression of cMET was more often identified in GC patients with deeper T (P= 0.016), nerve involvement (P = 0.006) and the Lauren diffuse type (P = 0.029). EGFR in cases with vessel permeation (P = 0.012), and HER-2 in cases of distant metastasis (P =0.031), non-nerve involvement (P = 0.024), the Lauren intestinal type (P < 0.001) and G1/G2 grade (P < 0.001). Conclusion The receptors tyrosine kinase (RTKs) markers of cMET, EGFR and HER-2 might involve in the advance of GC, such as invasion and metastasis. The expression status of them could be used as the risk prediction and even the basis for future personalized therapy, especially for tyrosine kinase inhibitor (TKI) therapy, for GC patients.

Follow-up Results and Clinical Significance of Magnetic Resonance Spectroscopy Combined with Fluorescence Microscopy in Resection of High Grade Supra-tentoria Gliomas / 中国医学科学院学报

Objective To investigate the clinical effectiveness of magnetic resonance spectroscopy (MRS) combined with sodium fluorescein(FL) in the treatment of high grade gliomas(HGG). Methods From August 2013 to 2015 November,the clinical data of 72 supratentorial HGG(WHO grade Ⅲ-Ⅳ) patients who had received surgical treatment in our hospital were retrospectively studied,among whom 43 cases received MRS combined with intra-perative FL navigation(observation group),and 29 cases only received conventional surgery(control group). Post-operative radiotherapy and chemotherapy were applied for more than 3 months. Routine enhanced MRI were performed 24-48 hours after the operation to investigate the extent of tumor resection. Six months after the operation,the quality of life of patients was evaluated by using the Karnofsky score,and 1-year postoperative survival rate and progression-free survival(PFS) were observed. Results Postoperative MRI showed that the rate of gross total resection(GTR) in observation group was significantly higher than that in control group(72.09%vs.51.72%;χ=23.88,P=0.001),and the GTR rate of WHO grade Ⅳ tumors was significantly higher than that of WHO grade Ⅲ tumors in observation group(92.86% vs.62.07%;χ=6.06,P=0.042). The postoperative Karnofsky score in the observation group was significantly higher than that in control group(μ=2.34,P=0.021). The mean time of follow-up was(16.4±2.4) months(8-21 months) and there was no statistical significant difference between observation group and control group in 1-year survival rate(74.07% vs.77.50%;χ=4.90,P=0.165) and PFS [(13.2±1.2) months vs.(12.7±2.0) months;χ=7.26,P=0.067]. In observation group,the PFS of WHO grade Ⅳ patients was significantly higher than that in control group [(14.2±0.3) months vs.(10.0±1.1) months;χ=11.03,P=0.031]. There was also no statistical significant difference between WHO grade Ⅳ tumors in two groups in terms of 1-year survival rate(71.43% vs.72.54%;χ=5.33,P=0.089),and there was no statistical significant difference between WHO grade Ⅲ tumors in two groups in 1-year survival rate(75.86% vs. 72.22%;χ=3.78,P=0.250) and in PFS [(13.7±1.4) months vs.(12.4±0.8) months;χ=4.85,P=0.083]. Conclusions MRS combined with intraoperative FL navigation technology can improve the resection rate and improve survival quality of patients,and there is no evidence that MRS combined with intraoperative FL navigation prolong the overall survival of patients with high-grade gliomas. Different outcome may be found with longer follow-up and increased simple size.

Effect of brucine on metabolism of osteoblasts and osteoclasts in multiple myeloma / 中国实验血液学杂志

This study was aimed to explore the influence of brucine on the early differentiation of osteoblasts and the metabolic pathway of osteoclast in multiple myeloma (MM) and to compare the effects of brucine and bortezomib on MM. The half inhibitory concentration (IC(50)) of brucine and bortezomib on MM cell line U266 was determined by MTT method; the mRNA levels of alkaline phosphatase (ALP), osteocalcin (OC), osteoprotegerin (OPG) and osteoprotegerin ligand (RANKL) were detected by RT-PCR after the supernatant of cultured U266 cells was added into the culture system for inducing the differentiation of osteoblast line MC3T3-E1 and culturing. The results showed that the IC(50)of bortezomib and brucine on U266 cells for 48 hours were 22.4 nmol/L and 0.16 mg/ml respectively. As compared with osteoblasts treated by supernatant of cultured MM cells alone, the mRNA levels of ALP, OC and OPG in osteoblasts treated by brucine combined with supernatant of cultured MM cells were enhanced (p < 0.05), while the RANKL mRNA level was lowered (p < 0.05), moreover the enhanced and lowered degree also was large (p < 0.05). It is concluded that the influence of brucine on metabolism of osteoblasts and osteoclasts in MM may be realized through the regulation of osteoclasts by osteoblasts. The therapeutic efficacy of brucine on MM is superior to bortezomib.

Effects of survivin siRNA on growth, apoptosis and chemosensitivity of ovarian cancer cells SKOV3/DDP / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore a new approach in gene therapy of ovarian cancer, we used RNAi to inhibit survivin gene expression, and explore the effect of survivin and neu RNAi on growth, apoptosis and chemosensitivity of ovarian cancer cell line SKOV3/DDP cells.</p><p><b>METHODS</b>Expression vector of survivin gene-targeting siRNA was constructed using pSilencer 1.0-U6 vector containing U6 promotor (pSilencer-survivin) and transfected into SKOV3/DDP cells by lipofectamine. The untransfected group and pSilencer-control group were used as control. The expressions of survivin mRNA and protein were identified by RT-PCR and Western blot assay. The proliferation of SKOV3/DDP cells was determined by MTT assay. The apoptosis rate and cell cycle distribution were analyzed by flow cytometry. Cisplatin (DDP) resistance experiment was performed in SKOV3/DDP cells with RNAi.</p><p><b>RESULTS</b>Survivin RNAi plasmid knocked-down survivin expression in SKOV3/DDP cells obviously, arrested the cells at G(1)/G(0) phase, inhibited the cell proliferation and promoted cell apoptosis. The IC(50) of DDP to SKOV3/DDP cells transfected with survivin siRNA was dropped.</p><p><b>CONCLUSION</b>Survivin RNAi can partly suppress the expression of survivin in SKOV3/DDP cells, inhibit the cell proliferation and promote cell apoptosis. Survivin RNAi can enhance the cell sensitivity to apoptosis induced by cisplatin, which implies that survivin RNAi may partly reverse the drug resistance of ovarian cancer. RNAi could be a new approach for gene therapy of cancer.</p>

Differential expression of genes related to transcription in cultured hepatoma cells with intact genome of hepatitis C virus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome.</p><p><b>METHODS</b>Huh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed.</p><p><b>RESULT</b>HCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening.</p><p><b>CONCLUSION</b>Intact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.</p>

Establishment of rat hypophyseal compression model and changes in its postoperative biological characteristics / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a rat model of hypophyseal compression and observe and analyze the changes in its biological characteristics after operation.</p><p><b>METHODS</b>The rats were subjected to compression of the pituitary gland by stuffing the autologous muscular tissue into the hypophyseal fossa. The postoperative mortality of the rats was recorded and the volume of the hypophyeseal fossa, body weight, daily food intake, water intake, urine volume and urine specific gravity were measured.</p><p><b>RESULTS</b>Rat models of the hypophyseal compression model were successfully established by this procedure, which resulted in an increase of the volume of hypophyseal fossa by 35%. Rapid body weight loss occurred within 5 weeks after the operation (by as much as 31% on day 10). The rats exhibited recovery of appetite after 2 weeks, but their food intake was still less than that in the control group. Manifestations of central diabetes insipidus occurred gradually, which were especially obvious at 2 weeks and persisted afterwards, and at this time point, significant increment of urine volume (55.4-/+15.9 vs 18.5-/+5.8 ml) and lowered urine gravity (1.011-/+0.004 vs 1.036-/+0.006) were observed.</p><p><b>CONCLUSION</b>Rat models of hypophyseal compression can be established successfully by the described procedure, and the compression results in alteration of the rats' metabolic behaviors, which may differ from the effects of hypophysectomy and damage of pituitary stalk.</p>

Expression of survivin and its significance in esophageal cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the expression of survivin in esophageal cancer and elucidate its function in esophageal cancer.</p><p><b>METHODS</b>Expression of surviv in was detected in paired normal and tumor tissues from patients with esophageal cancer by semi-quantitative RT-PCR. A dominant-negative survivin (surT34A) was transfected into esophageal cancer EC9706 cells (EC9706surT34A). Colony formation and apoptosis of the parental and surT34A-transfected EC9706 cells were examined in soft agar and by flow cytometry, respectively.</p><p><b>RESULTS</b>Survivin mRNA expression of tumor tissues was higher than normal tissues in 18/27 (66.7%) samples. The expression level of survivin mRNA in tumor tissues (2.08 +/- 1.32) was significantly higher than that in normal tissues (1.22 +/- 1.09). EC9706 surT34A cells formed fewer colonies on agar than the non-transfected ones. After serum withdrawal, EC9706surT34A had higher apoptotic ratio than control, but survivin could reduce the apoptotic ratio.</p><p><b>CONCLUSION</b>Overexpression of survivin is a common eventin esophageal cancer. The dominant-negative survivin can partially inhibit the malignant phenotype of esophageal cancer.</p>

Long term survival results in advanced prostate cancer treated with combined androgen blockade / 中华男科学杂志

<p><b>OBJECTIVE</b>To understand long-term survival rate after combined androgen blockade (CAB) in patients with advanced prostate cancer.</p><p><b>METHODS</b>A selected population of 59 patients with advanced prostate cancer were treated with CAB. 28.81% (17/59) of patients had clinical locally advanced disease (stage T3-4N0M0), and 45.76% (27/59) of patients had metastatic disease (stage TxNxM+). Overall, patients were followed for a median of 62 (range 6-136) months.</p><p><b>RESULTS</b>Of the 59 patients with advanced prostate cancer, 3-year, 5-year and 7-year overall survival rates were 79.36%, 61.46% and 49.15%, respectively. The 5-year survival rate were 80.77% and 32.65% for clinical locally advanced disease and metastatic disease. Specifically, men with poorly differentiated prostate cancer had a 5-year survival of only 30% when compared with men with well-differentiated prostate disease who had a 5-year survival of 86.21%.</p><p><b>CONCLUSION</b>Based on these findings, men with poorly differentiated cancer, stage T3c-4NxMx or TxNxM+ and PSA level above 30 microg/L had a high probability of dying from their advanced prostate cancer.</p>

Suppression of c-myc expression by interference RNA in HepG2 hepatocellular carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect of RNA interference (RNAi) on c-myc expression in hepatocellular carcinoma cell line, HepG2.</p><p><b>METHODS</b>Expression vector of c-myc gene-targeting small interference RNA (siRNA) was constructed (psilencer-c-myc) and transfected into HepG2 cells by lipofectamine, and the unloaded vector was used as control (mock). The expression of c-myc mRNA and protein was identified by quantitive PCR and Western blot. Apoptosis of the transfected cells was examined by flow cytometry and immunofluorescent microscopy.</p><p><b>RESULTS</b>After HepG2 cells were transfected with psilencer-c-myc, the expression of c-myc mRNA and protein was suppressed with an inhibition rate of 67% compared with the mock-transfected cells. Apoptosis was identified in the transfected HepG2 cells.</p><p><b>CONCLUSION</b>The expression of c-myc at transcriptional and translational levels in HepG2 cells transfected with siRNA is markedly inhibited, which may be associated with the induction of apoptosis.</p>