Orbicularis oculi muscle resection combined with orbicularis oculi muscle shortening and lower eyelid retractor reduction in the treatment of elderly lower eyelid entropion / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the efficacy of orbicularis oculi muscle resection combined with orbicularis oculi muscle shortening and lower eyelid retractor reduction in the treatment of elderly lower eyelid entropion.METHODS:A retrospective study was conducted among 97 elderly patients(175 eyes)with lower eyelid entropion who admitted to the hospital from June 2019 to June 2021. According to the treatment method, the patients were divided into control group(47 patients of 82 eyes treated with orbicularis oculi muscle resection)and combination group(50 patients of 93 eyes treated with orbicularis oculi muscle resection combined with orbicularis oculi muscle shortening and lower eyelid retractor reduction). The two groups were compared in terms of short-term curative effect, perioperative indexes, scores of symptoms and signs before and after surgery, width of palpebral fissure before and after surgery, direction of eyelashes, exposure rate of lacrimal caruncle, complications, and patient satisfaction.RESULTS: The total response rate in the combination group was significantly higher than that in the control group(95% vs 80%, P=0.004). The intraoperative blood loss, operation time and hospital stay of the combination group were significantly more/longer than those of the control group(P<0.001). The scores of symptoms and signs such as lacrimation, foreign body sensation, photophobia and irritation in the combination group after the surgery were significantly lower than those in the control group(all P<0.001). After surgery, the width of palpebral fissure, direction of eyelashes and exposure rate of lacrimal caruncle in the combination group were higher than those in the control group(P<0.001). The incidence of postoperative complications in the combination group was lower than that in the control group(8% vs 18%, P=0.032). The patient satisfaction scores of comfort level, trichiasis correction, scar appearance, eyes symmetry and appearance in the combination group were higher than those in the control group(all P<0.001).CONCLUSION: Orbicularis oculi muscle resection combined with orbicularis oculi muscle shortening and lower eyelid retractor reduction is effective and safe in the treatment of elderly lower eyelid entropion, which can meet the requirements of the patients.

Analysis of norovirus surveillance in adult infectious diarrhea in Putuo District of Shanghai / 上海预防医学

ObjectiveTo investigate the prevalence and epidemiological characteristics of norovirus in adult cases with infectious diarrhea in Putuo District of Shanghai， and provide scientific evidence for the prevention and control of infectious diarrhea caused by norovirus. MethodsFecal samples， clinical information and epidemiological data were collected from January 2013 through December 2019 in surveillance hospitals in Putuo District of Shanghai. Norovirus was examined by real-time fluorescent quantitative reverse transcription polymerase chain reaction （rRT-PCR）. ResultsIn 1 389 adult cases with infectious diarrhea， norovirus positive rate was 25.41%， which was significantly higher in male （27.16%） than female （23.89%）. Furthermore， in 353 cases positive for norovirus， GⅡ group was the most common （77.98%）.The positive rate was highest in the cases aged 30-44 years. Spring， autumn and winter were the seasons with higher incidence of norovirus （September to May）. The norovirus-infected cases had more nausea， vomiting， hyperactivity of bowel sounds， and watery stool， compared to the negative cases （P<0.05）. ConclusionThe detection rate of norovirus remains high in adult cases with infectious diarrhea in Putuo District of Shanghai， of which GII is predominant. Seasonality may be spring， autumn and winter. Therefore， it warrants the countermeasures， such as surveillance and health education， for prevention and control of norovirus in susceptible population during epidemic seasons.

Analysis of norovirus surveillance in adult infectious diarrhea in Putuo District of Shanghai / 上海预防医学

ObjectiveTo investigate the prevalence and epidemiological characteristics of norovirus in adult cases with infectious diarrhea in Putuo District of Shanghai， and provide scientific evidence for the prevention and control of infectious diarrhea caused by norovirus. MethodsFecal samples， clinical information and epidemiological data were collected from January 2013 through December 2019 in surveillance hospitals in Putuo District of Shanghai. Norovirus was examined by real-time fluorescent quantitative reverse transcription polymerase chain reaction （rRT-PCR）. ResultsIn 1 389 adult cases with infectious diarrhea， norovirus positive rate was 25.41%， which was significantly higher in male （27.16%） than female （23.89%）. Furthermore， in 353 cases positive for norovirus， GⅡ group was the most common （77.98%）.The positive rate was highest in the cases aged 30-44 years. Spring， autumn and winter were the seasons with higher incidence of norovirus （September to May）. The norovirus-infected cases had more nausea， vomiting， hyperactivity of bowel sounds， and watery stool， compared to the negative cases （P<0.05）. ConclusionThe detection rate of norovirus remains high in adult cases with infectious diarrhea in Putuo District of Shanghai， of which GII is predominant. Seasonality may be spring， autumn and winter. Therefore， it warrants the countermeasures， such as surveillance and health education， for prevention and control of norovirus in susceptible population during epidemic seasons.

Comparative Study of Transrectal Real-time Tissue Elastography and Magnetic Resonance Diffusion-Weighted Imaging in the Diagnosis and Guided Biopsy of Prostate Cancer / 中山大学学报(医学科学版)

@#【Objectives】 To comparatively analyze the diagnostic performance of transrectal real- time elastography（TRTE）and magnetic resonance diffusion- weighted imaging （DWI）in differentiating benign from malignant prostatic lesions，to evaluate the value of the two methods in guided prostate biopsy，and to investigate the correlation between the two methods and Gleason scores. 【Methods】 A total of 126 patients with suspected prostate cancer underwent prostate biopsy. Preoperative tests of TRTE and DWI were performed in all of the included patients. Combined with pathological results，the diagnostic efficacy of TRTE and DWI for prostate cancer and the effects of prostate biopsy guided by the two methods were compared ，and the relationship between the elastography score and ADC value and Gleason scores were also evaluated.【Results】 The sensitivity，specificity，accuracy of diagnosing prostate cancer by TRTE were 78.8% ，78.3% ，78.6% ，the sensitivity，specificity，accuracy of diagnosing prostate cancer by DWI were 87.9% ，90% ，88.9% ， there was no significant difference of sensitivity and specificity between the two groups（P > 0.05），and the accuracy was statistically different（P < 0.05）. The AUC of elastography score and ADC value were 0.859 and 0.906，the accuracy of the diagnosis of benign and malignant prostate lesions by ADC value method was higher than elastography score ，but there was no statistically significant difference （P > 0.05）. A significant positive correlation was found between elastography score and Gleason scores，while a significant negative correlation was found between ADC value and Gleason scores.【Conclusions】TRTE and DWI is valuable in diagnosis of prostatic lesions. Biopsy guided by the two methods can improve the detection rate of prostate cancer and can provide indicative evidence for tumor differentiation analysis.

Application of superb microvascular imaging in manifesting the blood flow of the penile cavernous artery in vascular ED patients / 中华男科学杂志

Objective@#To investigate the application value of superb microvascular imaging (SMI) in the diagnosis of penile vascular ED.@*METHODS@#Seventy-two ED patients underwent SMI and color Doppler flow imaging (CDFI), all ultrasonographically diagnosed with penile vascular ED. We compared SMI and CDFI in detecting the grades of blood flow in the cavernous artery and the lengths of time needed to obtain satisfactory blood flow spectrum from the patients.@*RESULTS@#SMI mainly revealed grades Ⅲ and Ⅳ blood flow, in 43 and 20 of the 72 patients (87.5%), while CDFI chiefly manifested grades Ⅰ and Ⅱ blood flow, in 26 and 32 cases respectively (80.6%). The former showed significantly better manifestations of the penile cavernous artery than the latter. It took less time to obtain the spectrums of grades Ⅲ and Ⅳ blood flow (［1.52 ± 0.18］ and ［1.21 ± 0.11］ min) than grades Ⅰ and Ⅱ (［5.23 ± 0.44］ and ［4.46 ± 0.65］ min), and SIM took significantly less time than CDFI (［1.32 ± 0.42］ vs ［4.53 ± 0.67］ min, P < 0.05).@*CONCLUSIONS@#SMI is superior to CDFI in better manifesting the blood flow of the penile cavernous artery and shortening the examination time, and therefore deserves a wide application in the diagnosis of vascular ED.

Polysaccharide extracts of Astragalus membranaceus and Atractylodes macrocephala promote intestinal epithelial cell migration by activating the polyamine-mediated K channel / 中国天然药物

Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.

Polysaccharide extracts of Astragalus membranaceus and Atractylodes macrocephala promote intestinal epithelial cell migration by activating the polyamine-mediated K channel / 中国天然药物

Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.

Application of transvaginal ultrasound in diagnosis and treatment of cesarean scar pregnancy / 上海交通大学学报（医学版）

Objective · To investigate the performance of transvaginal ultrasound (TVS) on cesarean scar pregnancy (CSP), and evaluate the value of TVS combined with MRI in the diagnosis and treatment of CSP. Methods · A retrospective analysis was made in 42 patients with CSP in Shanghai General Hospital in recent 3 years. The TVS performance, MRI results, and clinical data of the patients were analyzed. Results · In all 42 cases, 36 were diagnosed as CPS. TVS missed one case, and misdiagnosed 5 cases, which diagnostic accuracy was 85.7%. Thirty-six cases were tested by MRI, therein 32 cases were diagnosed correctly, 3 cases missed, 1 case misdiagnosed, which diagnostic accuracy rate was 88.9%. Among the 36 CSP cases, 30 were simple sac type and 6 were mixedmass type. For the simple sac type, different treatment was chosen according to the muscle thickness of the lower uterine segment incision measured by TVS. When it was more than 3 mm, suction surgery was used. But when it was no more than 3 mm, MRI examination was further used, and the surgical resection or uterine artery embolism+MTX+suction surgery was chosen. Conclusion · Both TVS and MRI have a high diagnostic accuracy for CSP, and TVS combined with MRI can provide important reference for CSP treatment options.

Application of transvaginal ultrasound in diagnosis and treatment of cesarean scar pregnancy / 上海交通大学学报（医学版）

Objective · To investigate the performance of transvaginal ultrasound (TVS) on cesarean scar pregnancy (CSP), and evaluate the value of TVS combined with MRI in the diagnosis and treatment of CSP. Methods · A retrospective analysis was made in 42 patients with CSP in Shanghai General Hospital in recent 3 years. The TVS performance, MRI results, and clinical data of the patients were analyzed. Results · In all 42 cases, 36 were diagnosed as CPS. TVS missed one case, and misdiagnosed 5 cases, which diagnostic accuracy was 85.7%. Thirty-six cases were tested by MRI, therein 32 cases were diagnosed correctly, 3 cases missed, 1 case misdiagnosed, which diagnostic accuracy rate was 88.9%. Among the 36 CSP cases, 30 were simple sac type and 6 were mixedmass type. For the simple sac type, different treatment was chosen according to the muscle thickness of the lower uterine segment incision measured by TVS. When it was more than 3 mm, suction surgery was used. But when it was no more than 3 mm, MRI examination was further used, and the surgical resection or uterine artery embolism+MTX+suction surgery was chosen. Conclusion · Both TVS and MRI have a high diagnostic accuracy for CSP, and TVS combined with MRI can provide important reference for CSP treatment options.

Development of a cell-based peroxisome proliferator-activated receptors (PPARs) screening model and its application for evaluation of triterpenoids isolate from Alismatis Rhizoma / 中国中药杂志

Peroxisome proliferators activated receptors (PPARs) are closely related to human chronic disease, such as diabetes mellitus and the other metabolic diseases. In this study, a cell-based PPARs (PPAR α/β/γ) model was developed for the screening of PPARs agonists from Alismatis Rhizoma (AR). Firstly, 293T cells were transfected with the reconstructed plasmid pBind-PPAR (α, β, or γ)-LBD and reporter gene pGL4.35, and the known PPARs agonists were used as the positive control (fenofibrate for PPARα, L165041 for PPARβ, and rosiglitazone for PPARγ). The ability of activation for PPARs was evaluated by analyzing the expression value of luciferase. Afterward, the 14 pure triterpenoids isolate from AR were analyzed on the developed PPARα, PPARβ and PPARγ screening assay method. The results showed that the compounds 5, 6, 7, 8, 13 and 14 from AR have the ability of activation for PPARα. The compounds 5 and 7 from AR have the ability of activation for PPARβ. The compounds 6, 7, 8 and 12 from RA have the ability of activation for PPARγ.In this study, the compound 12 from AR were found to display significant activation on PPARγ for the first time. AR triterpenoids extracts had the ability of activation for PPARα, PPARβ and PPARγ. The results suggested that triterpenoids extracts from AR were PPARα, PPARβ and PPARγ agonists. The results will help to provide reference for clinical application of AR, and establish a model for PPARs on 293T cell, which can be used to screen and evaluate PPARs natural agonists.

The Neuro-Protective Effect of the Methanolic Extract of Perilla frutescens var. japonica and Rosmarinic Acid against H2O2-Induced Oxidative Stress in C6 Glial Cells

Neurodegenerative diseases are often associated with oxidative damage in neuronal cells. This study was conducted to investigate the neuro-protective effect of methanolic (MeOH) extract of Perilla frutescens var. japonica and its one of the major compounds, rosmarinic acid, under oxidative stress induced by hydrogen peroxide (H2O2) in C6 glial cells. Exposure of C6 glial cells to H2O2 enhanced oxidative damage as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and thiobarbituric acid-reactive substance assays. The MeOH extract and rosmarinic acid prevented oxidative stress by increasing cell viability and inhibiting cellular lipid peroxidation. In addition, the MeOH extract and rosmarinic acid reduced H2O2-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the transcriptional level. Moreover, iNOS and COX-2 protein expression was down-regulated in H2O2-indcued C6 glial cells treated with the MeOH extract and rosmarinic acid. These findings suggest that P. frutescens var. japonica and rosmarinic acid could prevent the progression of neurodegenerative diseases through attenuation of neuronal oxidative stress.

Synthesis and immunogenicity indentification of artificial antigen of ginsenoside Rg1 / 中草药

Objective: To synthesize the artificial antigen of ginsenoside Rg1-bovine serum albumin (Rg1-BSA) and the artificial coated antigen of ginsenoside Rg1-polylysine (Rg1-PLL), and to provide the basis for the preparation of monoclonal antibody (MAb) and the establishment of immunoassay method. Methods: Rg1-BSA and Rg1-PLL were synthesized by sodium periodate oxidation method. The characterization of the synthesis was examined by UV spectrometry and TLC method. The titer and specificity of the antibody in serum of immunised mice were detected by indirect enzyme-linked immunosorbent assay (I-ELISA) and indirect competitive enzyme-linked immunosorbent assay (I-CELISA), respectively. Results: According to the UV and TLC, the Rg1 was successfully conjugated with BSA and PLL. I-ELISA and IC-ELISA methods were developed using Rg1-PLL. The anti-Rg1 antibody obtained from immunized mice could bind to Rg1 specially and the titer was up to 1:80000. Conclusion: The artificial immunogen Rg1-BSA and coated antigen Rg1-PLL are successfully synthesized, which could be used to prepare the MAb of Rg1 and establish the immunoassay method.

Effect of DNA methyltransferase 1 and methyl-CpG-binding protein 2 abnormal expression on cervical lesions and related interaction / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the effect of DNA methyltransferase 1 (DNMT1) and methyl-CpG-binding protein 2 (MeCP2) on cervical cancer and cervix precancerous lesion.</p><p><b>METHODS</b>74 patients with cervix squamous cell carcinoma(SCC), 52 patients with cervical intraepithelial neoplasm I (CIN I), 60 patients with cervical intraepithelial neoplasm II - II (CIN II-III)and 58 patients with histologically diagnosed cervix inflammation(CI), were included in this study. Information as demography, reproductive history, life style, HPV infection were collected. Western Blot were used to detect the expression of DNMT1 protein and MeCP2 protein. Real-time PCR was used to detect the expression of DNMT1 and MeCP2 mRNA.</p><p><b>RESULTS</b>Levels of DNMT1 and MeCP2 protein expression increased gradually with the deterioration of cervical lesion (H = 94.33, P < 0.001;F = 21.580, P < 0.001). Along with the deterioration of cervical lesion, levels of DNMT1 and MeCP2 mRNA expression were gradually increasing( F = 4.758, P = 0.003; F = 7.804, P < 0.001). Data from Correlation analysis showed that both protein (r = 0.287, P < 0.001) and mRNA(r = 0.179, P = 0.005)were positive correlated with DNMT1 and MeCP2.</p><p><b>RESULTS</b>of our study indicated that there was an additive interaction between high-expression of DNMT1 protein and high-expression of MeCP2 protein in SCC or CIN II-III. However, there was an additive interaction between high-expression of DNMT1 mRNA and high-expression of MeCP2 mRNA in SCC or CIN II-III.</p><p><b>CONCLUSION</b>Results from our study revealed the fact that both high expression of DNMT1 protein and high expression of MeCP2 protein could increase the risk of cervix cancerization. According to our findings, there might be a synergistic action existed between DNMT1 and MeCP2 during the progression of cervix cancelation.</p>

Effect of folate in modulating the expression of DNA methyltransferase 1 and methyl-CpG-bingding protein 2 in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effects of folate on the expression of DNA methyltransferase 1 (DNMT1) and methyl-CpG-bingding protein 2 (MeCP2) in cervical cancer cell lines.Methods Experimental study was carried out in vitro.Human cervical cancer cell lines,including C33A cell with HPV negative and Caski cell with HPV16 positive,were treated with different concentration of folate.The expression of DNMT1 and MeCP2 protein (by Western blot)and mRNA (by real-time PCR) were then detected in the two cell lines.Results It was found that supplement of folate was able to reduce the cell proliferation in C33A cell (r=0.984,P＜0.001) and Caski cell (r=0.978,P=0.002),as well as induced the cell apoptosis (C33A:r=0.989,P＜0.001 ;Caski:r=0.994,P＜0.001).Results showed that the expression levels of DNMT1 protein (C33A:r=-0.914,P＜ 0.001 ; Caski:r=-0.859,P=0.003) and MeCP2 protein (C33A:r=-0.830,P=0.005 ;Caski:r=-0.981,P＜0.001) decreased gradually with the increase of folate concentrations,but the expression of DNMT1 and MeCP2 mRNA was not observed in Caski or C33A cell.When at the same levels of folate,the expression of DNMT1 protein or mRNA was higher in Caski cell than in C33A cell.However,the expression of MeCP2 protein or mRNA was higher in C33A cell than in Caski cell.Conclusion Our fimding indicated that adequate foleta could effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,thus would reverse the aberration protein expression of DNMTl and MeCP2.That there might be a synergistic action between HPV16 infection and parafunction of DNMT l in cervical cancer,being noticed.

TTV and HPV co-infection in cervical smears of patients with cervical lesions in littoral of Zhejiang province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of transfusion-transmitted virus (TTV) and human papillomavirus (HPV) co-infection in cervical smears of patients with cervical lesions in littoral of Zhejiang province and analysis of transmitted route.</p><p><b>METHODS</b>Nested polymerase chain reaction (nPCR) was established. TTV DNA were tested by nPCR in cervical smears of 95 patients with cervical lesions and 55 healthy women, paired serum samples were available from 55 and 42 women, and their viral titer. The genotypes of 95 specimens of cervical cytology were detected with HybriMax. The phylogenetic group of TTV was determined by means of nPCR with N22 primers.</p><p><b>RESULTS</b>The prevalence of TTV DNA in cervical smears of patients with cervical lesions and healthy women was 52.7% (29/55) and was comparable with that in paired serum sample (50%). Symptomatic women had significantly higher prevalence of TTV DNA in cervical smears (74.7%) than healthy controls (P = 0.005). The TTV DNA prevalence in patient serum samples was 51%. The phylogenetic groups of TTV serum isolates were concordant with those of TTV from cervical smears of the same subjects, and genotype was G1b. The TTV viral titer in cervical smears were 10 to 1000 times as high as in serum. The total infection rate of HPV was 98.9% in patients, and was 27.3% in healthy women. The frequently detected genotype was HPV16, 18, 33 of HSIL, and HPV6 of LSIL. The HPV positive study subjects had significantly higher TTV DNA prevalence than HPV negatives (P = 0.02).</p><p><b>CONCLUSION</b>High prevalence of TTV in cervical smears suggests that sexual transmission is another mode of expansion of TTV infection among the population. The higher viral titer in cervical smears than in the respective serum samples might indicate active TTV replication in the female genital tract. Nevertheless, cooperation between TTV and HPV needs to be further investigated.</p>

Co-location of ACh-sensitive BK channels and L-type calcium channels in type II vestibular hair cells of guinea pig / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the mechanisms of the influx of calcium ions during the activation of ACh-sensitive BK channel (big conductance, calcium-dependent potassium channel) in type II vestibular hair cells of guinea pigs.</p><p><b>METHODS</b>Type II vestibular hair cells were isolated by collagenase type IA. Under the whole-cell patch mode, the sensitivity of ACh-sensitive BK current to the calcium channels blockers was investigated, the pharmacological property of L-type calcium channel activator-sensitive current and ACh-sensitive BK current was compared.</p><p><b>RESULTS</b>Following application of ACh, type II vestibular hair cells displayed a sustained outward potassium current, with a reversal potential of (-70.5 +/- 10.6) mV (x +/- s, n = 10). At the holding potential of -50 mV, the current amplitude of ACh-sensitive potassium current activated by 100 micromol/L ACh was (267 +/- 106) pA (n = 11). ACh-sensitive potassium current was potently sensitive to the BK current blocker, IBTX (iberiotoxin, 200 nmol/L). Apamin, the well-known small conductance, calcium-dependent potassium current blocker, failed to inhibit the amplitude of ACh-sensitive potassium current at a dose of 1 micromol/L. ACh-sensitive BK current was sensitive to NiCl2 and potently inhibited by CdCl2. NiCl2 and CdCl2 showed a dose-dependent blocking effect with a half inhibition-maximal response of (135.5 +/- 18.5) micromol/L (n = 7) and (23.4 +/- 2.6) micromol/L (n = 7). The L-type calcium channel activator, (-) -Bay-K 8644 (10 micromol /L), mimicked the role of ACh and activated the IBTX-sensitive outward current.</p><p><b>CONCLUSION</b>ACh-sensitive BK and L-type calcium channels are co-located in type II vestibular hair cells of guinea pigs.</p>

Cloning and expression of Oreochromis aurea gonadotropin-releasing hormone cDNA in Escherichia coli and its immunogenicity for mice / 生物工程学报

To study the function of the GnRH protein, the recombinant pMAL-GnRH was constructed and expressed in TB1 E. coli. The cDNA encoding gonadotropin-releasing hormone (GnRH) and GnRH associated peptide (GAP) was amplified from total RNA of O. aurea pituitary glands by reverse transcription polymerase chain reaction (RT-PCR), and then blasted against other GnRH cDNA sequences in the GenBank. The analysis of the sequence data indicated that the coding region of the cDNA fragment, which encoded 89 amino acid residues, was about 400 bp in size. The amplified cDNA fragment was cloned into the prokaryotic expression vector, pMAL-c2x, to produce the expression vector pMAL-GnRH. The recombinant plasmid was transformed into E. coli TB1. GnRH-MBP fusion protein was obtained after the addition of IPTG into the growth media. SDS-PAGE analysis revealed that the GnRH-MBP was expressed after induction with IPTG for 4 h. A protein band of 56 kD appeared on SDS-PAGE gel and was proved by Western blot. The mass production of the recombinant protein was about 41.6% of total bacteria protein. After purification and cleavage of the fusion protein purified GnRH protein could be obtained. Then the fusion protein was used to immunise some ICR mice to produce anti-GnRH antibody. This fusion protein could significantly elicit specific antibody response in immunized mice compared with the blank groups, and the titers against GnRH reached peak 0.707 +/- 0.320 at the 5th week after immunization. These results demonstrated that recombinant protein could induce high GnRH antibody responses in laboratory animals.

Prediction of the Secondary Structure and B Cell Epitopes of DMO and DMT Protein in Oreochromis aureus / 中国生物工程杂志

The secondary structure of the protein of DM0 and DMT in Oreochromis aureus were predicted by the methods of Garnier-Robson, Chou-Fasman and Karplus-Schulz based on the amimo acid sequences of DM0 and DMT. And Hydrophilicity plot, Surface probability and Antigenic index for DM0 and DMT protein were obtained by the methods of Kyte-Doolittle, Emini and Jameson-Wolf, respectively. Combined the results according to these methods, the B cell epitopes for DM0 and DMT protein were predicted. The results demonstrated that there were some centers of?-helix in the DM0 protein' s N- terminal No. 80 - 112, 144 -147, 193- 194, 251 - 255, 260 - 269 and No. 279- 283, and in the DMT protein' s N-terminal No. 61 -86, 98 - 105, 140 - 146, 239 -241 and No. 269 -273. And there are some centers of?-sheet in the DM0 protein' s N-terminal No. 59 -61, 69 -70, 148 - 150 and No. 383 -390, and in the DMT protein's N-terminal No. 125 -129, 207-213, 255-264 and No. 281-284. Furthermore, the DMO protein' s N-terminal No. 40-41,44 -45, 50-51, 128-129, 189-192, 204-207, 216-222, 226-233, 244-246, 298 - 299 and No. 323 -326, and the DMT protein' s N-termianl No. 12 - 13, 26 - 27, 43 - 44, 58 - 60, 93 - 95, 115 - 120, 136 -139 and No. 149 -151 may be the flexible regions. Moreover the B-cell epitopes possibly localized in or nearby the DMO protein's 1 -5, 41 -51, 65-67, 86-89, 98-110, 154-170, 183-203, 205 -248, 258-264, 284 - 291, 293 - 298, 270 - 375, 389 - 392 and No. 402 - 410, and DMT protein' s N-termianl No. 1 - 9, 17 - 28, 77 - 84, 114 - 123 , 131 - 139, 157 - 184 and No. 96 - 207. Theses results are helpful for studies on sex control mechanism of DMO and DMT in Oreochromis aureus.