Polymerase chain reaction-based assays facilitate the breeding and study of mouse models of Klinefelter syndrome / 亚洲男科学杂志(英文版)

Klinefelter syndrome (KS) is one of the most frequent genetic abnormalities and the leading genetic cause of nonobstructive azoospermia. The breeding and study of KS mouse models are essential to advancing our knowledge of the underlying pathological mechanism. Karyotyping and fluorescence in situ hybridization are reliable methods for identifying chromosomal contents. However, technical issues associated with these methods can decrease the efficiency of breeding KS mouse models and limit studies that require rapid identification of target mice. To overcome these limitations, we developed three polymerase chain reaction-based assays to measure specific genetic information, including presence or absence of the sex determining region of chromosome Y (Sry), copy number of amelogenin, X-linked (Amelx), and inactive X specific transcripts (Xist) levels. Through a combined analysis of the assay results, we can infer the karyotype of target mice. We confirmed the utility of our assays with the successful generation of KS mouse models. Our assays are rapid, inexpensive, high capacity, easy to perform, and only require small sample amounts. Therefore, they facilitate the breeding and study of KS mouse models and help advance our knowledge of the pathological mechanism underlying KS.

The progress in clinical management of difficult decannulation after ureteral stent retention / 医学研究生学报

The long-term retention of ureteral stent (double J tube) leads to the displacement and fracture of double J tube, and the formation of peritube stones, which are the main causes of the difficult decannulation through conventional cystoscopy. Its clinical treatment is more complex, involving different minimally invasive endoscopic techniques, and even by traditional open surgery. In recent years, more and more reports on the difficulty of removing double J tubes after retention. Among them, the multi-mirror combined operation method has been recommended, and the KUB scoring system based on imaging examination contributes to evaluate the difficulty of operation and prognosis of patients before operation. This article reviews the diagnosis of double J tube retention, the causes of difficult decannulation, preoperative preparation and progress of surgical management.

Rapid screening for Klinefelter syndrome with a simple high-resolution melting assay: a multicenter study / 亚洲男科学杂志(英文版)

Klinefelter syndrome (KS) is the set of symptoms that result from the presence of an extra X chromosome in males. Postnatal population-based KS screening will enable timely diagnosis of this common chromosomal disease, providing the opportunity for early intervention and therapy at the time point when they are most effective and may prevent later symptoms or complications. Therefore, through this study, we introduced a simple high-resolution melting (HRM) assay for KS screening and evaluated its clinical sensitivity and specificity in three medical centers using 1373 clinical blood samples. The HRM assay utilized a single primer pair to simultaneously amplify specific regions in zinc finger protein, X-linked (ZFX) and zinc finger protein, Y-linked (ZFY). In cases of KS, the ratios of ZFX/ZFY are altered compared to those in normal males. As a result, the specific melting profiles differ and can be differentiated during data analysis. This HRM assay displayed high analytical specificity over a wide range of template DNA amounts (5 ng-50 ng) and reproducibility, high resolution for detecting KS mosaicism, and high clinical sensitivity (100%) and specificity (98.1%). Moreover, the HRM assay was rapid (2 h per run), inexpensive (0.2 USD per sample), easy to perform and automatic, and compatible with both whole blood samples and dried blood spots. Therefore, this HRM assay is an ideal postnatal population-based KS screening tool that can be used for different age groups.

Forensic data analysis on 34 death cases of medical disputes due to aortic dissecting aneurysm rupture / 复旦学报（医学版）

Objective To investigate the reasons for the death of patients,the relationship of medical disputes with their death by aortic dissecting (AD) aneurysm (known as AD) ruptures during diagnostic and therapeutic procedures and the judgement for contribute ratios.Methods A total of 34 sudden death cases due to AD were collected from 2001 to 2016 due to medical tangle and underwent forensic pathological analysis.Clinical data were checked according to the results of forensic pathology.The reasons for the death of patients and the relationship of medical disputes with their death and contribute ratios of medical disputes were analyzed according to literature reports.Results In the 34 cases,23 cases were from medical institutions at the country level,and 11 cases were from municipal medical institutions.All patients had other diseases,in whome 27 cases had definite hypertension,31 cases died of cardiac tamponade,and 3 cases of hemorrhagic shock.The pathological analysis showed that aorta cystic in the middle necrosis,coronary atherosclerosis and coronary artery stenosis (grade Ⅰ-Ⅳ).Fifteen cases had coronary and aortic atherosclerosis,and 1 case had pulmonary artery dissection.All cases were not given definite clinical diagnosis and effective treatment for AD.Therefore,there was cause-and-effect relationship between clinical procedures of misdiagnosis and mistreatment and the death of patients.Conclusions The pathogenesis of AD is complicated,and a lack of awareness in grass-roots doctors leads to the misdiagnosis and mistreatment easily primary care doctor.In medical disputes of sudden death induced by AD,clinical procedures of misdiagnosis and mistreatment lead to indirect causal relationship with the death of patients.

Opportunity of forensic medical identification for simple traumatic subarachnoid hemorrhage / 复旦学报（医学版）

Objective To investigate the identification time and attention issue for simple traumatic subarachnoid hemorrhage (STSAH).Methods A total of 51 cases of STSAH from Center of Forensic Science,Siping Policy Security Bureau during 2014 to 2016 were analyzed retrospectively.Statistical methods used for sex,age,bleeding site,injury style and treatment outcome in forensic medical identification.Results In 46 cases of STSAH,high-density shadow in bleeding site narrowed gradually with regular change by CT scan,meanwhile clinical features disappeared gradually after longer treatment and was given the recognition in clinical identification.In addition,5 cases of STSAH had no obvious change 14 days after the injury confirmed by repeated CT,which were not given the recognition by forensic medical identification.Conclusions Early evaluation of injury degree in STSAH cases was carefully made to ensure the reliability of forensic medical identification;meantime false positive results could be avoided by follow-up CT scan.

Identification of origins of marker chromosomes using fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the value of fluorescence in situ hybridization (FISH) and bacterial artificial chromosome FISH (BAC-FISH) for the diagnosis for patients with marker chromosomes.</p><p><b>METHODS</b>Sixteen patients with marker chromosomes were analyzed with technologies including GTG-banding, Q-banding, multiplex FISH and BAC-FISH.</p><p><b>RESULTS</b>The marker chromosomes in the 16 patients were verified as der(Y) (2 cases), psu dic(Y) (1 case), psu dic(15) (1 case), dic(15) (1 case), del(Y) (1 case), r(X) (5 cases), i(14 or 22) (2 cases), i(18) (1 case).</p><p><b>CONCLUSION</b>FISH and BAC-FISH can both verify the origin of marker chromosomes and provide accurate information for the diagnosis and treatment of patients.</p>

Cigarette smoking decreases sperm nucleoprotein transition in infertile males / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the impact of cigarette smoking on sperm nucleoprotein transition and its association with sperm motility in infertile males.</p><p><b>METHODS</b>We examined the semen quality and sperm nucleoprotein transition of 116 non-smokers and 113 heavy smokers (aged 25 -50 years) who visited the Research Institute of Obstetrics and Gynecology for male infertility. We determined the rate of individual sperm nucleoprotein transition by aniline blue staining and analyzed the correlation of cigarette smoking with routine semen parameters and the rate of sperm nucleoprotein transition. Based on the smoking index (SI) derived from smoking frequency (no. of cigarettes/d) multiplied by smoking duration (yr), the men with SI = 0 were considered as non-smokers, and those with SI > or = 200 as heavy smokers.</p><p><b>RESULTS</b>The rate of abnormal sperm nucleoprotein transition was significantly higher in the asthenozoospermic (23.5 +/- 9.4, P < 0.01) and oligoasthenozoospermic (28.2 +/- 9.2, P < 0.01) than in the normozoospermic males (19.0 +/- 9.0). Compared with the non-smokers, cigarette smoking remarkably reduced sperm nucleoprotein transition in both the men with normal sperm motility (21.9 +/- 9.8 vs 16.8 +/- 7.7, P < 0.01) and those with abnormal sperm motility (26.0 +/- 9.9 vs 22.7 +/- 8.8, P < 0.05). A weak correlation was observed between the rate of sperm nucleoprotein transition and routine semen parameters.</p><p><b>CONCLUSION</b>Cigarette smoking is not significantly correlated with sperm motility but decreases sperm nucleoprotein transition in infertile males.</p>

Study on effects of kurarinol combined with glycyrrhizic acid on cellular immunity of patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore effects of kurarinol combined with Diammonium Glycyrrhizinate on specific cellular immunity of patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>Sixty-three CHB patients were randomly divided into two groups, 32 cases in group of kurarinol combined with Diammonium Glycyrrhizinate group (combined therapy group) were treated with 600 mg kurarinol glucose injection intravenously, once a day for one month, then 200 mg kurarinol capsule was used orally, three times a day for two months. 150 mg Diammonium Glycyrrhizinate for Injection was added to 250 ml 10% glucose injection for intravenous drip, once a day for one month, then 150 mg Diammonium Glycyrrhizinate capsule was used orally, three times a day for two months; 31 case in kurarinol group (single drug group) only used kurarinol, methods and dosage were the same as those of treatment group. HBV specific CTL, T cell subgroups, change of Th1 and Th2 level, HBV-DNA and HBeAg negative rate of the two groups were compared.</p><p><b>RESULTS</b>Three months after treatment, HBV specific CTL, CD4 + and Th1 of combined therapy group were higher than those before treatment, and higher than those of single drug group after treatment (P < 0.01).</p><p><b>CONCLUSION</b>HBV-DNA and HBeAg negative rate between the two groups had no statistic significance (P > 0.05).</p><p><b>CONCLUSION</b>Kurarinol combined with Diammonium Glycyrrhizinate can further increase HBV specific CTL, CD4+ and Th1 level of CHB patients.</p>

Multiplex ligation-dependent probe amplification for detecting AZF microdeletions on the Y chromosome in infertile men with azoospermia or severe oligozoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the possibility of applying multiplex ligation-dependent probe amplification (MLPA) to the detection of azoospermia factor (AZF) microdeletion on the Y chromosome in infertile men with azoospermia or severe oligozoospermia.</p><p><b>METHODS</b>DNA samples were obtained from 147 azoospermia or severe oligozoospermia patients and 154 normal controls. After denatured at 95 degrees C, the samples were hybridized to the specific probes designed for the AZF region. With the ligase, the hybrid products were amplified by a pair of universal primers labeled with FAM fluorescence, and then separated by capillary electrophoresis for data analysis. Meanwhile all the samples were subjected to multiplex-PCR (mPCR) analysis for sequence-tagged sites (STS) in the AZF region.</p><p><b>RESULTS</b>STS deletion was detected in 22 (15.0%) of the 147 patients but not in the normal controls. By MLPA, 40 (27.2%) of the patients were found with specific probe omission in the AZF region, as compared with 20 cases in the control group.</p><p><b>CONCLUSION</b>Compared with mPCR, MLPA has a better sensitivity in detecting AZF microdeletions, and it provides more precise genetic information on the AZF regions, which may contribute to in-depth exploration into the etiological mechanism of impaired spermatogenesis.</p>

Genetic counseling and clinical outcome of fetus with de novo chromosomal aberrations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the chromosome rearrangements and clinical outcome in fetus detected at prenatal diagnosis, and provide information for genetic counseling about de novo chromosomal aberrations.</p><p><b>METHODS</b>From January 2006 to December 2009, we found 12 cases of de novo chromosomal aberrations in 2 583 cases of prenatal cytogenetic analyses and reviewed the karyotypes, other experimental analyses data, fetal ultrasound findings and clinical outcomes.</p><p><b>RESULTS</b>Out of the 12 de novo chromosomal aberrations, 10 had unbalanced translocations and 2 had balanced reciprocal translocations. Eight of the 10 unbalanced translocation cases were terminated therapeutically, and 2 were delivered with full term. Neonates were phenotypically normal in the 2 cases with unbalanced translocations, but 1 had language retardation when followed up. The two balanced translocation cases were delivered with full term, and the neonates were phenotypically normal and clinical examinations were normal too.</p><p><b>CONCLUSION</b>Detailed cytogenetic and molecular study will be adjunctive tools for predicting the phenotype of fetus with de novo chromosomal aberrations. Fetal ultrasound examination will provide convincible demonstration to determine the outcome of pregnancy.</p>

Relationship between HBeAg seroconversion with genotypes and HBV specific CTL in patients with chronic hepatitis B treated with Adefovir dipivoxil / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore relationship between HBeAg seroconversion with HBV genotypes and HBV specific CTL in patients with chronic hepatitis B (CHB) treated with Adefovir dipivoxil.</p><p><b>METHODS</b>Seventy CHB patients had positive HBV DNA (HBV DNA > or = 1 x 10(4) copy/ml), 45 cases had positive HBeAg, of whom 23 cases (51. 11%) had genotype B, 22 cases (48.89%) had genotype C. ALT > 2 x upper limit of normal value (ULN), human leukocyte antigen (HLA)-A(n) positive, patients were treated with Adefovir dipivoxil (commercial name is Mingzheng, Zhengda Tianjing Pharmaceutical Company), 10 mg, orally, once a day. After treatment for 12 months, observe relationship between HBeAg seroconversion with HBV genotypes and HBV specific CTL.</p><p><b>RESULTS</b>After treatment with Adefovir dipivoxil for 12 months, HBV specific CTL (0.68% +/- 0.11%) was higher than that before treatment (0.33% +/- 0.11%), t = 8.36 P < 0.001, HBV DNA (3.01 +/- 0.2) log10 copy/ml was lower than that before treatment (6.27 +/- 0.70) log10 copy/ml, t = 12.63 P < 0.001, HBV DNA turned negative (< 500 copy/ml) 43 cases (61.43%), in 45 cases with positive HBeAg, HBeAg turned negative in 13 cases (28.89%), 8 cases had HBeAg seroconversion (17.78%), HBV specific CTL (0.86% +/- 0.05%) of patients with HBeAg seroconversion is higher than (0.61% +/- 0.07%) of patients without HBeAg seroconversion (37 cases, 82.22%) t = 7.88, P < 0.001. In 8 cases with HBeAg seroconversion, 7 cases had genotype B (30.43% of genotype B), 1 cases had genotype C (4.55% of genotype C), chi2 = 5.15, P < 0.05.</p><p><b>CONCLUSION</b>Adefovir dipivoxil can enhance HBV specific cellular immunity of CHB patients. After treatment, occurrence of HBeAg seroconversion is related to increase of HBV specific CTL level and may be related to genotypes.</p>

Influence of adefovir dipivoxil on HBV specific CTL in patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the influence of adefovir dipivoxil on HBV specific CTL in patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>10 mg adefovir dipivoxil (Zhengda Tianjing Pharmaceutical Company) was used for CHB patients with positive HBV DNA (HBV DNA > or = 1 x 10(4) copies/ml), ALT > 2 x upper limit of normal value (ULN) and positive human leucocyte antigen (HLA)-A2, orally, once a day for 3 months. Real time fluorescent quantitative PCR was used to determine HBV DNA and flowcytometer was used to determine HBV specific CTL.</p><p><b>RESULTS</b>After treatment with adefovir dipivoxil for 3 months, HBV specific CTL (0.52 +/- 0.11)% was higher than that before treatment (0.34 +/- 0.14)%, t = 6.78 P < 0.01, HBV DNA of 28 cases turned to negative (<1 x 10(3) copies/ml) (62.22%). HBV DNA of 17 cases failed to turn negative 3 months after treatment, but their HBV DNA level was lower [(4. 18 +/- 0.4) log 10 copies/ml] than that before treatment [(6.23 +/- 0.73) log 10 copies/ml], t = 9.99, P < 0.01.</p><p><b>CONCLUSION</b>Adefovir dipivoxil can improve HBV specific cellular immunity in patients CHB.</p>

Application of fluorescence in situ hybridization in prenatal diagnosis of complex chromosomal abnormalities / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the application of fluorescence in situ hybridization (FISH) technique in prenatal diagnosis of complex chromosomal abnormalities.</p><p><b>METHODS</b>Eleven prenatal diagnosis cases (8 from amniocentesis and 3 from cord blood) with complex chromosomal abnormalities detected by routine G-banding, were further analyzed by FISH.</p><p><b>RESULTS</b>The FISH technique confirmed the results of balanced chromosome rearrangements detected by G-banding, and clarified the structure of the derivative chromosomes in the 3 amniocentesis samples and the origin of the mark chromosomes in the 2 cord blood samples.</p><p><b>CONCLUSION</b>FISH can be used to diagnose the complex chromosomal abnormalities accurately in prenatal diagnosis, and can provide very useful genetic information for clinical diagnosis and treatment.</p>

Noninvasive prenatal diagnosis of single gene disorders through cell-free fetal DNA in maternal blood / 中华医学遗传学杂志

The discovery of cell-free fetal DNA (cff-DNA) in maternal plasma offered a new way to noninvasive prenatal diagnosis for single gene disorders. In the past decade, many techniques such as real-time PCR, pyrophosphorolysis-activated polymerization, mass spectrum and digital PCR have been developed for noninvasive prenatal diagnosis. In this review, the author discuss the principles, applications, advantages and disadvantages of these techniques.

Molecular cytogenetic studies of 25 males with azoospermia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical correlation of chromosome abnormalities and microdeletion in azoospermic factor (AZF) region on Y chromosome in 25 patients with azoospermia.</p><p><b>METHODS</b>Chromosome analyses were performed by using chromosome GTG-banding, Q-banding, fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) for AZF region on chromosome Yq.</p><p><b>RESULTS</b>Seven cases showed abnormal chromosome karyotype (28%). In 8 azoospermic patients tested, 2 showed microdeletions of AZFb (SY127, SY134)+ AZFc (SY254, SY255) and AZFc(SY243, SY158) on chromosome Yq, respectively.</p><p><b>CONCLUSION</b>Chromosome abnormalities and AZF microdeletion are major cause of azoospermia leading to male infertility; male with azoospermia and infertility should be referred to cytogenetic diagnosis by using chromosome GTG-banding, Q-banding after ruling out clinical factors including testopathy, obstructive azoospermia, and abnormalities in incretion and immune system. FISH or PCR analysis for AZF region on chromosome Yq should be done for the patient with azoospermia if Q-banding indicates the deletion above Yq12 region. It is of essential importance to provide precise diagnosis in genetic counseling for further clinical treatment.</p>

The Exploration and Practice of Environmental Microbiology Teaching Reform / 微生物学通报

Environmental Microbiology is an important basic course of Environmental Engineering.Its characteristic is content broad,quick development and strong practicality,thus this curriculum's teaching has certain degree of difficulty.Some suggestions and concrete measures about teaching reform,which included curriculum's course content,teaching method,experiment teaching and assessment methods were proposed in this paper.