Objective To establish a stable HCV full-length genome replication cell model which is labeled with reporter gene and easyly to quantify intracellular HCV proteins and RNA level. Methodsneo gene was inserted into Luc-JC1 to make Luc-JC construct. Luc-JC RNA was obtained by in vitro transcription and then delivered into Huh7 cells by transfection. G418-resistant clones of Huh7 cells were obtained by selection. Clones of HCV full-length genome replication cell were confirmed by luciferase activity assay, Western blot and cleaveage of eYFP-MAVS by HCV NS3/4A protease. Then, HCV replication cell colonies were treated by different dose IFN-α in order to observe the change of luciferase activity, HCV protein and RNA level. Results At 3-4 weeks post-transfection, visible colonies were selected and stained by crystal violet. Luciferase activity and HCV NS3, NS5A protein were detected by luciferase activity assay and Western blot, respectively. Subcellular localization of eYFP-MAVS transferred from mitochondria to cytoplasms by cleavage of NS3/4A protease in cell colonies. Luciferase activity, HCV protein and RNA diminished obviously after IFN-α treatment. Conclusion A stable HCV full-length genome replication cell model labeled by reporter gene was successfully established and reporter activity can be used to indicate level of HCV proteins and RNA in cells. This cell model is a useful tool for the study on HCV pathogenesis and the screening of antiviral drugs.

Objective Detrusor overactivity (DO) is one known cause of overactive bladder (OAB) symptoms in benign prostatic hyperplasia (BPH).In this study,OAB symptoms suggestive of DO in BPH were treated with α1-blocker monotherapy or α1-blocker and antimuscarinics add-on therapy,and the efficacy and safety were assessed.Methods BPH patients who suffered from OAB symptoms for at least 3 month were enrolled in a prospective self-control study from August 2010 to April 2012.The inclusion criteria are total international prostate symptom score (IPSS) ≥8,OAB Symptom Score (OABSS) ≥3,OABSS urgency score ≥2,Postvoid residual volume (PVR) ＜ 100 ml,and maximum urinary flow rate (Qmax) ≥ 5 ml/s.All the patients who met the inclusion criteria were treated with α1-blocker ( tamsulosin 0.2 mg/day or doxazosin 4 mg/day) for 2 weeks.After 2 weeks,patients with no symptomatic improvement ( OABSS≥3) underwent pressure-flow test,and those whose Pdet≥ 40 cm H2O and DO presented more than one time were added antimuscarinics (solifenacin 5 mg/day or tolterodine 4 mg/day) for an additional 2 weeks.OABSS,IPSS,QOL,Qmax and PVR were re-evaluated every 2 weeks.Results Ninety-four cases of BPH/OAB patients met the inclusion criteria and completed 4 weeks treatment.The baseline of total OABSS was 7.0 ± 1.3,IPSS was 17.0 ± 1.7,QOL was 5.0 ±0.7,Qmax was (8.8 ±2.5) ml/s and PVR was (86.0 ± 16.5) ml.After 2 weeks treatment with α1-blocker alone,OABSS was 5.2 ± 0.8,IPSS was 14.2 ± 1.9,QOLwas4.O±0.8,Qmaxwas (11.4±2.4) ml/s and PVR was (67.9±12.9) ml.After another2 weeks treatment with α1-blocker plus antimuscarinics,OABSS was 3.1 ± 0.8,IPSS was 11.1 ± 1.9,QOL was 3.1 ± 0.7,Qmax was ( 10.8 ± 2.4) ml/s and PVR was (71.8 ± 11.9 ) ml.Compared with baseline values,OABSS,IPSS,QOL,Qmax and PVR significantly improved (P ＜ 0.01 ) in α1-blocker monotherapy group and α1-blocker plus antimuscarinic group.The improvement of OABSS,IPSS,QOL scores of α1-blocker plus antimuscarinic group were greater than α1-blocker monotherapy group (p ＜ 0.05 ),while Qmax and PVR showed no differences between the two groups.No acute urinary retention (AUR) was deteted.Conclusion Both of α1-blocker monotherapy and α1-blocker with antimuscarinics add-on therapy can improve OAB symptoms.

Objective To evaluate the efficacy of mitomycin (MMC) intravesical chemotherapy for superficial bladder carcinoma by in vitro chemosensitivity using histoculture drug response assay (HDRA).Methods Forty-one cases of superficial bladder transitional cell carcinoma(TCC) were obtained,including 30 males and 11 females with mean age of 55 years.Of them,10 cases were Ta and 31 were T1 according to TNM stage system (UICC 2002) while 9 cases were G1,22 were G2 and 10 were G3 (WHO1973).All cases had no chemotherapy history before operation and were operated to retain bladder.Tumor specimens were cultured by three-dimensional histoculture.HDRA with im-proved MTT assay was utilized for chemosenstivity test of MMC with 1 g/L concentration and 2 hours exposure.Growth inhibition rate (GI) exceeding 70% was defined as high-sensitive while less than 50% GI was defined as insensitive,others were moderate-sensitive.All cases were performed standard intravesical chemotherapy with MMC 40 mg plus 40 mt saline.Every case was followed up every 3 months.The patients were followed up for 5 years or untill recurrence.Results The MMC chem-osensitivity was different among 41 patients.Thirteen cases were insensitive including 1 of Ta,12 of T1 (P=0.009) and 1 of G1,7 of G2,5 of G3(P=0.101).Total recurrence rate was 39%(16/41) af-ter 3 to 5 years follow-up.There were 1 of Ta,15 of T1 (P=0.059) and 10 of G2 6 of G3 (P=0.016).Insensitive group recurrence rate was 77% (10/13) while sensitive group was 21% (6/28,P= 0.004).Patients in sensitive group showed a longer median time(49.2 months) than patients in insen-sitive group (16.5 months,P<0.001) according to Kaplan-Meier analysis with Log-rank test.The MMC chemosensitivity was independent prognostic factor examed by Cox regression analysis (P= 0.008).There was a 78% correlation rate of chemosensitivity by HDRA to clinical effect of MMC in-travesical chemotherapy.Conclusion HDRA could evaluate MMC intravesical chemotherapy for su-perficial bladder TCC,improve therapeutic effect and lower tumor recurrence rate.

Objective To study the application of pediculated skin flaps in the treatment of com-plicated long urethratresia. Methods From March 1999 to May 2006, a total of 18 male patients with complicated long urethratresia were treated by using the pediculated skin flaps. The causes of urethratresia were 7 cases of postoperative pelvic fractures with posterior urethral stricture, 4 cases of transurethral intravesical chemotherapy, 3 cases of postoperative bulbar urethral stricture, 2 cases of gonorrhea, and 2 cases of long-time urethral catheter placement. Four cases were urethratresia nf cor-pus penis, 7 cases were anterior urethral obliteration, 7 cases were posterior urethral and anterior ure-thral obliteration. Urethro-perineal fistulas were found in 8 cases, posterior urethrorectal fistulas in 7 cases, false passage formations in 8 cases. The average length of urethratresia was 15.1 cm (range 8. 7 to 23. 0 cm). The urethral scar was rasected, the posterior urethrorectal fistula was repaired, and different kinds of pediculated skin flaps depending on the length of urethratreaia was used. Results All the patients were followed up for 12 to 18 months (mean 14 months). Fifteen patients voided well 3 months postoperatively, none of the urography showed stricture. The mean peak urinary flow rate was 16. 9 ml/s (range from 16. 5 to 21.7 ml/s). Of the other 3 cases, 1 case experienced difficult voi-ding due to the long and circuitous tabularized skin flap but recovered after proper shortening;1 case had restenosis for the infection of anastomosis but voided well after excision and reanastomnsis;1 casehad a urinary fistula resulting from hematoma and infection, but was successfully treated by the neo-plasty of the urinary fistula. The mean peak urinary flow rate was 17.0 ml/s (range 15.0 to 22.0 ml/s) for 17 patients 6 months postoperatively, except for one who experienced genuine urinary incon-tinence. At 9-18 months after operations, the mean peak urinary flow rate was 17.5 ml/s (range 15.8 to 22.5 ml/s) for 17 patients. Conclusion The single-stage urethroplasty based on pediculated skin flaps is a reliable and durable method for complicated long urethratresia.

Objective To use the cognitive reinforcement comprehensive intervention program constructed by our team to intervene in patients with spinal cord injury and evaluate its clinical application effect.Methods A non-randomized trial design was adopted to select 97 patients with spinal cord injury from November 2021 to September 2022.Forty-four patients from March to September 2022 in a Grade A hospital in Hefei City were included in the experimental group,and 53 patients from November 2021 to February 2022 were included in the control group.The cognitive reinforcement comprehensive intervention program was used to intervene in the experimental group,and the conventional rehabilitation nursing was used to intervene in the control group.The intervention lasted for 12 weeks in both groups.The Changsha Montreal Scale,Social Support Rating Scale,Rehabilitation Exercise Self-efficacy Scale,Spinal Cord Injury Independence Rating Scale and Hamilton Anxiety Scale were used to measure the two groups before intervention,1 month after intervention and 3 months after intervention.Results 40 cases in the experimental group and 48 cases in the control group completed the study.Repeated measurement ANOVA showed that the temporal,interactive and intergroup effects of cognitive function scores and anxiety scores were statistically significant(P＜0.05).The time effect and interaction effect of the subjective support dimension score,coping self-efficacy dimension score of the two groups were statistically significant(P＜0.05).One month after the intervention,the cognitive function scores of test group were higher than before intervention and control group,and the anxiety scores were lower than before intervention and control group(P＜0.05).Three months after the intervention,the scores of cognitive function,subjective support dimension and coping self-efficacy dimension of experimental group were higher than those before intervention and control group,and the scores of anxiety level were lower than those before intervention and control group(P＜0.05).Conclusion Comprehensive intervention of cognitive reinforcement can improve the cognitive function of patients with spinal cord injury,delay the process of cognitive impairment,enhance self-confidence,relieve anxiety,and promote physical and mental rehabilitation of patients.

Objective To study the relationship between hair selenium content and arsenism among population from endemic arsenism area caused by coal combustion in the southern of Shaanxi Province.Methods Four high-arsenic-exposed villages in southern Shaanxi Province were selected as the study area,and the nonarsenic-exposed villages nearby were selected as the control area,90 hair samples of residents exposed to high arsenic in the study area and 14 hair samples of residents in the control area were collected.Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to determine the contents of arsenic and selenium in hair samples.Selenium/hair arsenic (Se/As) value was calculated,and the relationship between selenium content in hair samples and arsenicosis was analyzed.Results The geometric mean of selenium concentration in hair from the study area was 8.72 mg/kg,which was significandy higher than that from the control area (0.54 mg/kg,t =9.811,P ＜ 0.05).However,selenosis case was not found in the study area.Selenium content in hair was not obviously correlated with arsenism (r =0.075,P ＞ 0.05),while the Se/As value was negatively associated with the grade of arsenism (P ＞ 0.05).The geometric mean of selenium content in hair of female from the study area was 12.79 mg/kg,which was higher than that in hair of male (6.5 mg/kg,t =1.738,P ＜ 0.01).Meanwhile,the Se/As value in hair of female from the study area (1.15) was significantly higher than that in hair of male (0.65,t =3.218,P ＜ 0.05).Conclusions The environment of the study area is polluted by arsenic and selenium derived from stone-coal burning.Antagonism between arsenic and selenium might reduce the toxicity of selenium.Therefore,selenosis is not found in the study area.In addition,antagonism between arsenic and selenium might also induce lower prevalence of arsenism for female in the studly area.

Objective To investigate the beneficial role of synbiotics in the intestinal microbiota of patients with chronic functional constipation (CFC).Methods According to the inclusion and exclusion criteria,6 patients with CFC were enrolled with their fresh fecal samples collected,after a continuous treatment of one month their fresh fecal samples collected again.Meanwhile,6 healthy volunteers were enrolled as the control group with their fresh fecal samples collected.All samples were transported with ice and stored in -80 ℃ refrigerator,and were analyzed by metagenomics sequencing.Results After 4 weeks of symbiotic treatment,the intestinal microbiota had changed in species in patients with CFC.Bacteria of Escherichia_ coli,Clostridium_ sp._ SS2/1 and Clostridium_ sp._ 7_ 3_ 54FAA,which were rich in the people with constipation,decreased in abundance after the treatment.Bacteria of Lactobacillus_ oris and Bifidobacterium _ animalis,which were rich in the healthy people,increased in content after the treatment.Bacteria of Veillonella_ parvula,Veillonella_ sp._ 6_ 1_ 27,Veillonella_ sp._ 3 _ 1_ 44 which were rich in the healthy people,decreased in content after the treatment.LEfSe analysis showed that Parabacteroides distaso nis,Escherichia_ coli and Enterobacter-cloacae were the specific species of the three groups respectively.Conclusion Synbiotics can change the intestinal microbiota showing therapeutic effect,thus can be used as a novel clinical treatment method.

Objective:To explore the effects of the transtheoretical model (TTM) -based intervention in patients undergoing coronary angiography (CAG) .Methods:Totally 146 patients who were scheduled to undergo CAG at Fuwai Central China Cardiovascular Hospital from May 2021 to May 2022 were selected by convenience sampling. Patients admitted from May to November 2021 were assigned to the control group ( n=73), and those admitted from December 2021 to May 2022 were assigned to the observation group ( n=73). The control group received routine care, while the observation group received TTM-based intervention. The effects of the intervention were evaluated using the Self-Rating Anxiety Scale (SAS), the Self-Rating Depression Scale (SDS), and a self-designed CAG compliance assessment form. Results:After the intervention, SAS and SDS scores decreased in both groups, with the observation group scoring significantly lower than the control group ( P< 0.05). The CAG compliance in the observation group was also significantly higher than that in the control group ( P<0.05) . Conclusions:The TTM-based intervention can improve the psychological state of CAG patients and enhance their compliance with CAG procedures.