Sexual behavior characteristics and influencing factors of gonorrhea patients in Yunnan Province / 中华疾病控制杂志

Objective To investigate the sexual behavior types, condom use and influencing factors of gonorrhea patients in Yunnan Province, and to provide evidence for the adjustment of sexually transmitted disease (STD) prevention and control strategy. Methods A cross-sectional study was carried out to investigate gonorrhea patients in 14 STD clinics in 7 more prevalent prefectures (cities) of Yunnan Province. A questionnaire survey was conducted to investigate the socio-demographic and sexual characteristics of the patients in a one-to-one way. Multiple Logistic regression model was used to analyze the influencing factors of sexual behavior. Results A total of 179 cases of gonorrhea were investigated. The average age was (29.01±8.93) years old. 95.53% (171/179) patients were 40 years old and 88.27% were male patients (158/179). Unmarried patients accounted for 55.31% (99/179). Service and self-employed patients accounted for 34.64% (62/179) and 23.46% (42/179) respectively. The rate of sexual intercourse with opposite sex was 98.32% (176/179), the rate of men who have sex with men was 1.68% (3/179) and the rate of having more than two sexual partners was 27.93% (50/179). The rate of sexual intercourse between temporary partners, spouses/fixed partners and commercial partners were 53.07% (95/179), 37.99% (68/179) and 8.94% (16/179) respectively. Recent sexual activity had lower condom use rate, 17.89% (17/95), 19.12% (13/68) and 18.75% (3/16) respectively. The main ways for gonorrhea patients to make temporary partners were friend introductions or gatherings, accounting for 66.32% (63/95). Condom use in male patients was worse than that in female patients (OR=0.234, 95% CI: 0.084-0.656, P=0.006). Conclusions The patients with gonorrhea were mainly unmarried young adults. The main risk behaviors were unprotected behaviors between heterosexual temporary and spouse/fixed sexual partners. The condom use consciousness was generally low. The promotion of safe sex education for young adults should be further strengthened, with special attention to the use of condoms for temporary sexual behavior and spouse/fixed sexual intercourse.

Quantitative Expression of RNA from Frozen Organs and Formaldehyde-fixed and Paraffin-embedded Tissues / 法医学杂志

Objective Quantitative analysis and comparison of the expression of ribonucleic acid （RNA） from frozen organs and formaldehyde-fixed and paraffin-embedded （FFPE） tissues. Methods Frozen specimens of human brain, myocardium and liver tissues as well as FFPE samples at different postmortem intervals were collected and mass concentration of RNA was extracted and detected. Reverse transcription-quantitative polymerase chain reaction （RT-qPCR） technology was used to analyze the amplification efficiency and relative expression of each RNA marker. Results The mass concentration and integrity of RNA extracted from FFPE samples were relatively low compared with frozen specimens. The amplification efficiency of RNA markers was related with RNA species and the length of amplification products. Among them, glyceraldehyde-3-phosphate dehydrogenase （GAPDH） and β-actin （ACTB） with relatively long amplification products failed to achieve optimal amplification efficiency, whereas 5S ribosomal RNA （5S rRNA） achieved ideal amplification efficiency and showed quite stable expression across various tissues, therefore it was chosen as internal reference marker. The expression quantity of GAPDH and ACTB in frozen specimens with longer postmortem intervals and in FFPE samples with relatively long amplification products was decreased. The expressions of tissue-specific microRNAs （miRNAs）, GAPDH and ACTB with relatively short amplification products had consistency in the same tissues and FFPE samples. Conclusion Through standardizing the RT-qPCR experiment, selecting the appropriate RNA marker and designing primers of appropriate product length, RNA expression levels of FFPE samples can be accurately quantified.

Effect of I1imidazoline receptor on the expression and function of α2Aadrenergic receptor / 国际药学研究杂志

Objective To investigate the effect of I1imidazoline receptor(I1R)on the expression and function of α2Aadrener-gic receptor(α2AAR)at the cellular level.Methods After sequencing and enzymatic identification,the mouse I1R and α2AAR plas-mids were transfected into CHO cells,respectively.The radioligand receptor binding assay and flow cytometry were used to select sin-gle cell clones,and the CHO cell lines stably expressing the mouse I1R or α2AAR were established.The CHO cell line that stably ex-presses both the mouse I1R and α2AAR were also established by the same technology and strategy.Then,the radioligand receptor bind-ing assay was used to determine the affinity and expression of α2AAR. Further,the effects of I1R on the α2AAR expression and the α2AAR agonist dexmedetomidine(DEX)-induced extracellular regulated protein kinase(ERK)phosphorylation were evaluated by the Western blotting.Results After transfection of mouse I1R and α2AAR plasmids,CHO cells grew normally.In the saturation binding experiments of membrane proteins from the CHO cells that stably expressed α2AAR,the Kdand Bmaxvalues of 3H-RX821002 were(0.96 ± 0.24)nmol/L and(0.29 ± 0.03)nmol/g protein,respectively.The expression levels of I1R were significantly increased in both the CHO cells expressing I1R and the CHO cells co-expressing α2AAR and I1R(P<0.05,P<0.01),when the cells that express exogenous I1R or α2AAR alone were trasfected again with the I1R plasmids.Moreover,in the CHO cells that transfected both I1R and α2AAR sta-bly,the I1R expression upregulated the α2AAR expression(P<0.01),and further increased the ERK phosphorylation induced by DEX through activating α2AAR(P<0.01).Conclusion I1R could upregulate the α2AAR expression and the ERK phosphorylation in-duced by DEX through activating α2AAR in the CHO cells that express exogenous I1R and α2AAR.This study presents a groundwork for further exploration of the relationship between I1R and α2AAR at the molecular level in future.

Methodological research of RNA extraction and quantitative analysis of long non-coding RNA from formalin-fixed paraffin-embedded human brain specimens / 上海交通大学学报（医学版）

Objective·To compare the quality of RNA extracted from fresh and formalin-fixed paraffin-embedded (FFPE) brain tissues and to explore the long non-coding RNA (IncRNA) expression level.Methods·FFPE samples stored under various conditions and paired frozen brain tissues were collected and total RNA qualities were then detected.Amplification efficiency (AE) and expression stability of each RNA marker were calculated and analyzed based on real-time quantitative PCR.After selecting reference biomarkers,normalized △ Ct values of candidate makers within different amplicon size were measured to assess the possibility of lncRNA quantification in FFPE tissues.Results·The purity of RNA extracted from FFPE was relatively high,but the RNA integrity was lower than fresh samples.All biomarkers were successfully amplified and amplification efficiencies of long-chain RNA markers were correlated with amplicon sizes,sample treatment and preservation conditions,namely temperature and storage time.5S,miR-9 and miR 125b achieved optimal AE and showed quite stable expression in all specimens,therefore they were chosen as control markers.Compared with fresh samples,the △ Ct values of only 2 lncRNA (HAR1F and MALAT1-L,whose amplicon size were both higher than 200 bp,respectively) increased in the FFPE samples kept in 4 ℃,while in FFPE tissues kept in room temperature,increments of the △ Ct values were significant for most target genes except for short amplicon markers (＜60 bp),which showed consistently stable expression in all brain specimens.Conclusion·RNA integrity is affected by sample treatment and preservation conditions,but IncRNA expression levels in FFPE tissues can be accurately quantificated by using optimal amplicon sizes and considerable reference markers.

Preparation of a polyclonal antibody against human LYZL4 and its expression in the testis / 中华男科学杂志

Objective@#To prepare a polyclonal antibody against human lysozyme-like protein 4 (LYZL4) expressed in the prokaryotic system and identify the distribution of LYZL4 in the testis.@*METHODS@#The full-length cDNA of LYZL4 was cloned into the pET32a plasmid and the expression of the recombinant LYZL4 (rLYZL4) was induced by IPTG. The rLYZL4 was purified by Ni-NTA and chitin affinity chromatography respectively and its bactericidal activity was observed by bilayer agar plate diffusion assay. The purified rLYZL4 was used as an immunogen to generate the polyclonal antibody, followed by examination of the antibody titer by ELISA and its specificity by Western blot. The distribution of LYZL4 in human tissue, sperm and seminal plasma was identified and its subcellular localization in the testis was determined by immunohistochemistry.@*RESULTS@#rLYZL4 was expressed efficiently in the prokaryotic system and exhibited no bacteriolytic activity against M. lysodeikticus and E. coli. The anti-rLYZL4 polyclonal antibody could bind the recombinant protein with a high sensitivity and specificity. LYZL4 was identified in the testis, epididymis and sperm protein extracts and localized in the acrosomal region of round and elongating spermatids.@*CONCLUSIONS@#An anti-rLYZL4 polyclonal antibody was successfully prepared using the prokaryotic expression system. LYZL4 was detected in the acrosomal region of round and elongating spermatids, suggesting an association with the structure and function of the acrosome.

Correlation between RNA Expression Level and Early PMI in Human Brain Tissue / 法医学杂志

OBJECTIVES@#To explore the correlation between the expression levels of several RNA markers in human brain tissue and early postmortem interval （PMI）.@*METHODS@#Twelve individuals with known PMI （range from 4.3 to 22.5 h） were selected and total RNA was extracted from brain tissue. Eight commonly used RNA markers were chosen including β-actin, GAPDH, RPS29, 18S rRNA, 5S rRNA, U6 snRNA, miRNA-9 and miRNA-125b, and the expression levels were detected in brain tissue by real-time fluorescent quantitative PCR. The internal reference markers with stable expression in early PMI were screened using geNorm software and the relationship between its expression level and some relevant factors such as age, gender and cause of death were analyzed. RNA markers normalized by internal reference were inserted into the mathematic model established by previous research for PMI estimation using R software. Model quality was judged by the error rate calculated with estimated PMI.@*RESULTS@#5S rRNA, miRNA-9 and miRNA-125b showed quite stable expression and their expression levels had no relation with age, gender and cause of death. The error rate of estimated PMI using β-actin was 24.6%, while GAPDH was 41.0%.@*CONCLUSIONS@#5S rRNA, miRNA-9 and miRNA-125b are suitable as internal reference markers of human brain tissue owing to their stable expression in early PMI. The expression level of β-actin correlates well with PMI, which can be used as an additional index for early PMI estimation.

Expression of human lysozyme-like protein 6 in the male reproductive system / 中华男科学杂志

<p><b>Objective</b>To study the expression of human lysozyme-like protein 6 (LYZL6) in the male reproductive system and its physiological role.</p><p><b>METHODS</b>The recombinant P. pastoris strain was cultured and induced with methanol to express LYZL6, followed by purification using chitin affinity chromatography. The bactericidal activity of the recombinant LYZL6 was observed by bilayer agar plate diffusion assay, and then the recombinant protein was used as an immunogen to generate polyclonal antibodies, whose specificity was examined by ELISA. The distribution of LYZL6 in the human tissue and semen was identified by Western blotting and the subcellular localization in the testis was investigated by immunohistochemistry.</p><p><b>RESULTS</b>At pH 5.6, recombinant LYZL6 exhibited a high bacteriolytic activity against M. lysodeikticus. ELISA analysis showed that the anti-LYZL6 polyclonal antibodies could bind the recombinant protein with a high specificity. Western blot manifested the expression of LYZL6 in the testis and epididymis, higher in the former than in the latter. LYZL6 was also detected in the sperm protein extract, while protein bands were not observed in the seminal plasma. Immunodetection with a specific antiserum localized the LYZL6 protein in the late spermatocytes and round spermatids.</p><p><b>CONCLUSIONS</b>LYZL6 has a higher bacteriolytic activity under low pH condition and is bound to spermatozoa after secreted in the testicular epithelia, suggesting that LYZL6 could act as a potential hydrolase for carbohydrates in zona pellucida penetration.</p>

The ultrasound and endocrine profile and their correlations in obese and non-obese women with polycystic ovary syndrome / 中华医学超声杂志（电子版）

Objective To study the different ultrasonic features in patients of polycystic ovary syndrome (PCOS) with or without obesity based on body mass index (BMI), and to investigate whether certain hormonal factors correlate with ovarian morphology and blood flow, and to discuss the role of ultrasound combined with hormone test in the diagnosis of obese PCOS. Methods One hundred and five women with PCOS were recruited. Patients were divided into two groups according to BMI;obese PCOS group (OB-PCOS, n=32, BMI≥25 kg/m2) and non-obese PCOS (NOB-PCOS, n=73, BMI<25 kg/m2). The ultrasonic parameters of follicle number (FN), ovarian volume (Vol), resistance index (RI) of ovarian stromal blood, RI of uterine artery and serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), the ratio of luteinizing hormone and follicle-stimulating hormone (LH/FSH), progesterone (P), estradiol (E2), free testosterone (FT), prolactin (PRL), sex hormoe binding globulin (SHBG), fasting plasma glucose (FPG), fasting insulin (FINS), the extent of insulin resistance and hyperandrogenism (HOMA-IR) were measured and compared. The correlation of the ultrasonic parameters and hormonal factors were analyzed. Results The Vol of OB-PCOS group was significantly higher than NOB-PCOS group [(12.25±4.89) ml vs (10.73±2.30) ml, t=2.20, P < 0.05]. FN and uterine artery RI of OB-PCOS group had a rising trend and RI of ovarian interstitial was on a reducing trend compared with NOB-PCOS group. But the differences were not statistically significant. The levels of FINS and HOMA-IR in OB-PCOS group [(14.82±6.45) mU/L and (3.91±3.30)] were significantly higher than those in NOB-PCOS group [(8.04±4.57) mU/L and (1.64±1.20)] (t=4.87, 3.47, respectively, both P < 0.01). And FSH in NOB-PCOS group was significantly higher than OB-PCOS group [(5.95±1.91) U/L vs (4.65±1.88) U/L, t=-2.77, P<0.01]. In POCS patients, FN was significantly associated with LH/FSH (r=0.35, P<0.01), and FT (r=0.38, P<0.01). Vol was significantly associated with LH/FSH, BMI, HOMA-IR and FPG (r=0.27, P<0.05;r=0.25, P<0.05;r=0.40, P<0.01;r=0.32, P<0.01). RI of ovarian stromal blood flow was significantly associated with SHBG (r=0.28, P<0.05). In OB-POCS group, RI of uterine artery was significantly associated with PRL (r=-0.58, P < 0.05). Vol was significantly associated with HOMA-IR (r=0.47, P < 0.05). In NOB-POCS group, FN was significantly associated with LH/FSH (r=0.33, P<0.05), and FT (r=0. 56, P<0.05). Vol was significantly associated with FT (r=0.31, P < 0.05). Conclusion There are some differences in the ultrasound and endocrine parameters between obese and non-obese PCOS patients, and some correlations exist between them.

Effects of citalopram on electrophysiological properties of cardiac myocytes / 上海第二医科大学学报

Objective To investigate the influence of citalopram on the fast response action potential,slow response action potential,in vitro electrocardiogram(ECG) and in vivo ECG of cardiac myocytes,and explore its mechanism of adverse cardiac effects. Methods Conventional microelectrode technique was employed to record the fast and slow response action potentials of the isolated papillary muscles of guinea pigs.In vivo and in vitro ECG were recorded from anesthetized animals and Langendorff-perfused hearts,respectively. Results Citalopram could prolong the RR interval and QRS duration of in vivo ECG.The premature ventricular contraction and atrial ventricular block were induced by 12.5?10-6 mol/L citalopram.The maximum ascending velocity of 0 phase(Vmax),action potential amplitude(APA) and action potential duration(APD50 and APD90) were dose-dependently decreased by citalopram in the fast and slow response action potentials of guinea pigs,respectively. Conclusion Citalopram can inhibit sodium and calcium channels effectively,which may be the ionic mechanism that citalopram induces arrhythmia in the clinical practice.

PTEN induces anoikis through its phosphatase activity in hepatocellular carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanisms of tumor suppressor gene PTEN on the induction of anoikis of hepatocellular carcinoma SMMC-7721 cells.</p><p><b>METHODS</b>SMMC-7721 cells were transfected with GFP plasmids containing wild-type PTEN or phosphatase inactivating mutant PTEN (C124A-PTEN) in vitro; The PTEN expression and the phosphorylation levels of focal adhesion kinase (FAK) and protein kinase B (PKB/Akt) were detected by Western blotting; Flow cytometry assay and laser scanning confocal microscopy were used to analyze apoptosis in adherent and non-adherent cells.</p><p><b>RESULTS</b>Compared with the control, PTEN expression in the cells transfected with wild-type PTEN increased to 248%, while the phosphorylation level of FAK and Akt decreased 65.2% and 89.1%, respectively; and the anoikis percentage increased from 9.5% to 31.3%. In the cells transfected with C124A-PTEN, neither the phosphorylation of FAK and Akt nor the anoikis percentage had obviously changed, although the PTEN expression enhanced dramatically in comparison with the control.</p><p><b>CONCLUSION</b>Through its phosphatase activity, tumor suppressor gene PTEN can suppress the phosphorylation of FAK and Akt, and induce anoikis in hepatocellular carcinoma cells.</p>

PTEN induces apoptosis and up-regulates p53 expression in HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of tumor suppressor gene PTEN on apoptosis and protein expression of p53 in HepG2 cells, as well as to explore its mechanisms.</p><p><b>METHODS</b>HepG2 cells were transfected with GFP plasmids containing wild-type PTEN or G129E-PTEN and C124A-PTEN in vitro. Both the expression of wild-type p53 and the phosphorylation of protein kinase B (PKB/Akt) and focal adhesion kinase (FAK) were detected by Western blotting. Flow cytometry and confocal microscopy were used to analyze apoptosis of the transfected cells.</p><p><b>RESULTS</b>Compared with the control, the expression of phosphorylated FAK and phosphoylated Akt were down-regulated in HepG2 cells transfected with wild-type PTEN (-65%, -93%) and G129E-PTEN (-65%, -35%), whereas the apoptosis percentage increased to (19.8+/-1.2)% and (9.2+/-0.6)%, and p53 expression was up-regulated by 120% and 50%, respectively. However, in the cells transfected with C124A-PTEN, neither the phosphorylation of FAK and Akt nor the apoptosis percentage and p53 expression had changed.</p><p><b>CONCLUSION</b>PTEN can dephosphrylate FAK through its protein phosphatase activity, and suppress phosphorylation of Akt mainly through its lipid phosphatase activity. Consequently, it can induce apoptosis of HepG2 cells and up-regulate p53 expression.</p>

Studying the mechanism of heart rate slowing down after birth by the method of HRV power spectrum analysis / 中国应用生理学杂志

<p><b>AIM</b>To explore the neurological mechanism of heart rate slowing down after birth and to investigate if the heart itself participates in the process.</p><p><b>METHODS</b>With the method of HRV power spectrum analysis, the regulatory mechanisms of autonomic nerves over heart rate were studied on different age groups of human beings and rabbits. Similar experiments were carried out on isolated perfused rabbit hearts in order to study if the heart itself were involved in the process of heart rate slowing down as they grew up.</p><p><b>RESULTS</b>The ratio of parasympathetic tone to sympathetic one is significantly increased as a child or rabbit grows up while the heart rate becomes slower. The spontaneous beating of isolated rabbit heart also slows down as the rabbit becomes older.</p><p><b>CONCLUSION</b>Both of the nerve factor and heart itself are involved in the process of slowing down of heart rate as a child grows up.</p>

The sinus node itself also plays a role in heart rate slowing down during postnatal development / 生理学报

The slowing down mechanism of heart rate during growth of the body after birth was studied in isolated rabbit heart and sinus node (SN) preparation with Langendorff perfusion method, conventional microelectrode recording and perforated patch for recording pacemaker current I(f). The radioimmunoassay was also used to measure the concentration of cAMP within SN cells. The results indicate that without the influence of nervous and humoral factors, the spontaneous heart rate would also become slower as the rabbit grew older, which is due to the decrease of spontaneous depolarized rate of phase 4 in SN cells. The negative directed shift of the threshold potential of I(f) and the decrease in cAMP concentration within SN cells may be responsible for the phenomenon. The results obtained suggest that besides the nervous and humoral factors which influence the heart rate, the changes in automaticity of SN cell itself may take part in the slowing down process as the body grows up.