Objective To construct directional expressing cDNA library from normal testes in Chinese males for further studying the structure and functions of spermatogenesis related genes. Methods mRNA of normal testes from a Chinese male was isolated and the cDNA synthesized by reverse transcription.After being digested by SalI and NotI ,dscDNA was subcloned into expressing plasmid pSPORT1 directionally.Finally,the directional expressing cDNA library from normal testes in Chinese males was constructed. Results The transformation rate of the library was 7?10 5.Ten clones were chosen and analysed. Digested by SalI and NotI ,the fragments of 0.5～2.0 kb were obtained with cDNA inserted. Conclusions The present study indicates that the directional expressing cDNA library from normal testes in Chinese males is qualified to screen genes and is considered to be helpful for studying the structure and functions of spermatogenesis related genes.

Objective To conduct a systematic review on the safety and efficacy of enhanced recov -ery after surgery ( ERAS) in perioperative management of hepatectomy .Methods A literature search was conducted on databases which included the PubMed , Embase, Cochrane Library, Sinomed, Wangfang, VIP and CNKI for randomized controlled trials ( RCTs) on application of ERAS in patients after hepatectomy . The data collection ended in August 2015.A meta-analysis was performed using RevMan 5.3 software.Re-sults Eleven RCTs which included 1074 patients were selected into this study .There were 530 patients in the ERAS group and 544 patients in the control group .On Meta-analysis, when compared with the control group, the ERAS group had significantly shorter length of hospital day (WMD=-2.36, 95%CI: -3.19~-1.54 , P<0.05 ) , shorter time for functional recovery ( WMD=-2.30 , 95%CI: -3.77 ~-0.83 , P<0.05), lower total complication rate (RR=0.65, 95%CI:0.52~0.80, P<0.05), and significantly decreased rates of postoperative pulmonary infection (RR=0.36, 95%CI:0.14~0.91, P<0.05) and nausea and vomiting (RR=0.48, 95%CI:0.26~0.89, P<0.05).There were no significant differences between the two groups on the rates of postoperative bleeding , biliary fistula, abdominal infection, delayed incisional healing, wound infection and urinary tract infection (P>0.05).The ERAS group had significant-ly lower hospitalization cost (SMD=-1.61, 95%CI:-2.42~-0.80, P<0.05), but the differences between the two groups on mortality and re-admission rates were not significant (P>0.05).When compared with the control group , the drainage tube removal time ( WMD=-2.83 , 95% CI:-3.92~-1.76 , P<0.05), time to first mobilization (SMD=-2.34, 95%CI:-2.98~-1.70, P<0.05), time to first feeding ( SMD=-5.08 , 95%CI: -9.33~-0.83 , P<0.05) , time to passage of first flatus ( SMD=-3.60, 95%CI:-4.85~-2.34, P<0.05) in the ERAS group were significantly shorter , but there was no significant difference on the time to the first bowel motion ( P>0.05 ) .Conclusions ERAS in the peri-operative management of hepatectomy was safe and beneficial .

Objective To compare everolimus (EVR) with calcineurin inhibitor (CNI) minimization or withdrawal on renal function of liver transplant patients with standard CNI therapy.Methods A search was conducted on databases which included the PubMed,Embase and Cochrane library for randomized controlled trials (RCTs) comparing EVR with CNI with minimization or withdrawal (the EVR group) with standard CNI therapy (the standard CNIs group) on renal function of liver transplant patients.A metaanalysis was performed using RevMan 5.3 software.Results Five RCTs which included 1 264 patients were selected into this study.There were 790 patients in the EVR group and 474 patients in the standard CNIs group.On meta-analysis,the EVR group had significantly better renal function (SMD =0.36,95％ CI 0.09 ～ 0.64,P ＜ 0.05),but higher rates of infection (RR =1.37,95％ CI 1.08 ～ 1.74,P ＜ 0.05),dyslipidemia (RR =2.46,95％ CI 1.79 ～ 3.38) and leukopenia (RR =2.37,95％ CI 1.32 ～ 4.26).No significant differences were found on the mortality and the acute rejection rates between the two groups (all P ＜ 0.05).Conclusions EVR with CNI minimization or withdrawal after liver transplantation provided effective immunosuppression and improved patients' renal function.The treatment increased the rates of infection,dyslipidemia and leukopenia.

To determining the postmortem interval (PMI) through quantitative analysis of the DNA degradation of cell nucleus in human brain and spleen by using image analysis technique (IAT). The brain and spleen tissues from 32 cadavers with known PMI were collected, subjected to cell smear every 1 h within the first 5-36 h after death, stained by Feulgen-Van's staining, Three indices reflecting DNA in brain cells (astrocytes) and splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray (AG) were measured by employing the mage analysis instrument. The results showed that IOD and AOD declined and AG increased with the prolongation of dead time within 5-36 h. A correlation between the PMI and gray parameters (IOD, AOD and AG) was identified and the corresponding regression equation was obtained. The parameters (IOD, AOD and AG) were proved to be effective quantitative indicators for accurate estimation of PMI within 5-36 h after death.
Cell Nucleus/*pathology ; DNA Degradation, Necrotic ; Forensic Pathology ; Liver/*pathology ; Postmortem Changes ; Spleen/*pathology ; Time Factors

OBJECTIVETo verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 (CASK/Id1) pathway in fibroblasts of human keloid.

METHODSImmunofluorescence laser was used to confirm CASK and Id1 protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Id1 expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Id1 protein of keloid fibroblasts was tested by immunoprecipitation.

RESULTSCASK and Id1 protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Id1 were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0.658 +/- 0.024, which was lower than that in the normal control group (1.076 +/- 0.008, t = 11.159, P < 0.05). The expression of Id1 mRNA was 0.497 +/- 0.014, which was higher than that in the normal control group (0.307 +/- 0.017, t = 15.148, P < 0.05). The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0.057 +/- 0.006, which was lower than that in the normal control group (0.168 +/- 0.012, t = 13.524, P < 0.05); the expression of Id1 protein was 0.812 +/- 0.035, which was higher than that in the normal control group (0.368 +/- 0.031, t = 16.356, P < 0.05). The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Id1 precipitate. There was a natural binding of CASK and Id1 in keloid fibroblasts.

CONCLUSIONCASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

Cell Proliferation ; genetics ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; metabolism ; Fibroblasts ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction

Objective To determine the effectiveness of systolic blood pressure gradient between the right arm and the right leg(SBPG) tests in the diagnosis of neonatal aorta anomaly(AoA).Methods The SBPG of infants suspected of having critical congenital heart diseases were collected prospectively,who were admitted to Guangdong General Hospital from January 2013 to December 2015.The results of SBPG test were compared with those of echocardiography or cardiac computed tomography (golden standard).The rates of true positive,true negative,false positive,false negative were calculated under cutoff values of 5 mmHg(1 mmHg =0.133 kPa),10 mmHg,15 mmHg and 20 mmHg,respectively.Receiver operating characteristic (ROC) curves was used to compare tests of different cutoff and the areas under the ROC curve were also calculated.Results Among 664 enrolled infants,67 cases were confirmed by golden standard test.The systolic blood pressure in the right arm,the legs and SBPG in AoA group and non-AoA group were (88.0±20.4) mmHgvs.(73.4±9.3) mmHg (P＜0.01),(66.1 ±10.1) mmHg vs.(69.0 ±9.7) mmHg(P＞0.05) and (22.6±17.8) mmHgvs.(2.3 ±4.8) mmHg(P ＜0.01),respectively.In these patients,31 cases(46.3％),31 cases(46.3％),27 cases(40.3％) and 21 cases(31.3％) were diagnosed of AoA,and 36 cases(53.7％),36 cases (53.7％),40 cases(59.7％),and 46 cases (68.7％) were missed by SBPG tests of 5 mmHg,10 mmHg,15 mmHg and 20 mmHg,respectively (P ＜ 0.01).The rates of true negative among those groups were 94.1％,99.5％,99.7％ and 100.0％,and the areas under ROC curve were 0.656,0.722,0.695 and 0.657,respectively (P ＜ 0.01).Conclusions Almost half of AoA infants could be screened out by SBPG test.The cutoff of 10 mmHg could probably be used to screen potential AoA infants,with higher true positive rate and lower false positive rate.

Objective To evaluate the safety and efficacy of ultrasound-guided percutaneous microwave ablation of renal neoplasms.Methods Twenty-two patients with pathologically proven renal neoplasms(18 renal cell carcinoma,3 renal angiomyolipoma,1 oncocytoma)with diameters of 1.0 to 3.8 cm were treated by microwave ablation.Cooled-shaft needle antenna was percutaneously inserted into the tumors under ultrasound guidance.One antenna was used for tumors less than 2 cm,two antennas were used for tumors larger than 2 cm.One thermal couple was placed adjacent to the tumor monitoring temperature in real-time during ablation.Immediate treatment efficacy was assessed by contrast-enhanced ultrasound within 3 days after ablation.Short-term efficacy was assessed by contrast-enhanced CT and/or contrast-enhanced ultrasound at 1,3,6 months and every 6 months thereafter.Results Twenty tumors were completely ablated in a single session,2 tumors were completely ablated in 2 sessions.No complications occurred.No residual tumor or recurrence was observed during follow-up.Conclusions Ultrasound-guided pereutaneous microwave ablation appears to be a safe and effective technique for the management of renal neoplasms in selected patients.

Objective To develop a computer-assisted multiangular guiding system for interventional ultrasound.Methods A computer-assisted multiangular guiding system was developed which included a high-accuracy electromagnetic tracker and a passive arm with 5 degree-of-freedom.The electromagnetic tracker was used to track ultrasound probe and needle.The position of B-mode ultrasound image plane and needle were displayed in real-time in the reconstructed 3D ultrasound.The passive arm was used to assist needle delivering.A 10mm ball immersed in a transparent water tank and a formal acoustical prototype was punctured with random angle by using this system.The systematic precision was assessed.Results The whole progress was finished successfully,the ball was touched by the tip of the needle for all the occasions,the error of the system was less than 5mm.Conclusion The computer-assisted multiangular guiding system can get rid of the limitation of angle and make the operation flexible and accurate.

BACKGROUND: Salvianolic acid B can ease nerve injury and promote neurogenesis, but its effects on proliferation,apoptosis and differentiation of neural stem cells in the hippocampus remain unclear.OBJECTIVE: To study the effects of salvianolic acid B on proliferation, apoptosis and differentiation of rat hippocampal neural stem cells following oxygen-glucose deprivation.METHODS: Hippocampal neural stem cells were isolated from newborn Sprague-Dawley rats, and divided into six groups, five of which were cultured in an incubator containing anaerobic mixtures (1% O2, 5% CO2 and 94% N2) for 150minutes followed by treatment with different concentrations of salvianolic acid B (0, 5, 10, 20, 40 mg/L), respectively.After 4 days of intervention, MTT was used to detect cell proliferation. After 48 hours of intervention, flow cytometry was used to detect cell apoptosis in the hippocampus. After 5 days of culture, flow cytometry was performed to evaluate the percentage of cells positive for neuron-specific enolase and glial fibrillary acidic protein. Normally cultured cells acted as controls (normoxic group).RESULTS AND CONCLUSION: Compared with the normoxic group, the proliferation of neural stem cells was decreased significantly (P < 0.01) and the rate of apoptosis was increased in the oxygen-glucose deprivation group (P <0.01). After treatment with different concentrations of salvianolic acid B, the cell viability and the ratio of neurons in total cells were increased, and the ratio of astrocytes was decreased, especially in 20 and 40 mg/L groups (P < 0.01). In conclusion, these results suggest that salvianolic acid B alleviates adverse effects of oxygen-glucose deprivation on neural stem cell proliferation, differentiation and apoptosis.

BACKGROUND:Liquiritin has the protective and nutritive effects on neural stem cells. However, the effect of liquiritin on neural stem cells from the brain of mouse embryos remains unclear. OBJECTIVE:To investigate the effects of different concentrations of liquiritin on the proliferation of neural stem cells from the brain of mouse embryos. METHODS:Neural stem cells were separated from the embryonic brain of Kunming white mice at the gestational age of 14 days. The identification of embryonic neural stem cells was performed by immunocytochemistry method. The expression of neural stem cells-special genes was determined by qRT-PCR. The cell growth curve was drawn and proliferation of embryonic neural stem cells treated with 0, 1, 2, 4 or 8 g/L liquiritin for 48 hours was measured by MTT assay. RESULTS AND CONCLUSION:(1) When cultured at day 5, al individual neural stem cells gathered together into neurospheres; with the extension of time, the neurospheres were enlarged, and gathered together into larger cell masses. (2) Results from immunocytochemistry showed that all the floating neurospheres was nestin-positive. Data from qRT-PCR revealed a higher expression of nestin mRNA, but there was no expression of neuron-specific enolase and glial fibrillary acidic protein in the neural stem cells. (4) The growth of neural stem cells was slow at the beginning. After 2-3 days, the cell proliferation quickly entered the exponential phase. After 4 days, the cell proliferation gradually slowed down, and the overall cell growth entered into the platform period. (5) The cell proliferation after treatment with 2, 4 or 8 g/L liquiritin was faster than that in the control group (0 g/L). To conclude, 2-8 g/L liquiritin could increase the proliferation of neural stem cells from the brain of mouse embryos.