Objective:To develop a HCV combined HBV DNA-based therapeutic vaccine.Methods:The HBV core gene and HCV core cDNA were inserted into the eukaryotic expression vector with two multiple cloning sites mammalian expression vector under the CMV promoter and RSC promoter respectively, named pRSC-HBV/HCV. Cellular expression of pRSC-HBV/HCV was assessed by Western blot and immunofluorescent study following transfection into SP2/0 cells. The Balb/c mice were immunized by multiple sites intramuscular injection with pRSC-HBV/HCV and the immune responses were detected.Results:The 21 kD and 14 kD core protein were observed. Both anti-HBc Ab and anti-HCV core Ab were detected in all immunized mice.Conclusion:The investigation demonstrated that pRSC-HBV/HCV could express HBcAg and HCV core protein, humoral immune response could be detected in mice immunized with pRSC-HBV/HCV.

【Objective】To observe the therapeutic effect of Qi-strengthening and yin-nourishing,stasis-removing and regeneration-promoting therapy for clinical diabetic nephropathy(CDN)and to explore its mechanism.【Methods】Eighty CDN patients with Qi-yin deficiency and stasis blocking kidney collaterals were randomized into groups A and B at the proportion of 3∶1 by using random number table.Group B(n=20)was given routine western treatment involving decreasing blood glucose,blood pressure and blood lipid,taking good quality diet with low protein,regulating water-electrolyte disturbance and acid-base balance,and preventing and curing complicated infection.Group A(n=60)was treated with Qi-strengthening and yin-nourishing,stasis-removing and regeneration-promoting herbal medicine(mainly composed of Radix Astragali,Fructus Mori,Radix Scrophulariae,Radix Angelicae Sinensis,Radix Salviae Miltiorrhizae,Radix et Rhizoma Rhei,Stigma Maydis and Rhizoma Chuanxiong).After treatment,the therapeutic effect was compared in the two groups.Meanwhile,the changes of fasting blood glucose,renal kidney,24-hour urinary protein,blood rheology,blood lipid and fibrinogen(FIB),as well as plasma endothelin 1(ET-1),tumor necrosis factor ?(TNF-?)and interleukin 1(IL-1)were observed.【Results】Qi-strengthening and yin-nourishing,stasis-removing and regeneration-promoting herbal medicine relieved the symptoms and signs of the patients,decreased 24-hour urinary protein content,and improved the renal function parameters in renal compensatory and decompensatory stages.The herbal medicine could also decrease blood lipid and fasting blood glucose levels,improve blood rheology indexes,and decrease serum IL-1 and ET-1 levels(P

Objective Serum pharmacochemistry was performed to screen the bioactive constituents of Resina Draconis.Methods Based on HPLC fingerprints of Resina Draconis,the migrating constituents absorbed into blood were determined by comparing the HPLC fingerprints of extraction of Resina Draconis(ERB),herb serum sample,and control serum sample,and with the help of LC-MS/MS.Results Six compounds absorbed into blood were detected,five of them were original in form which were 3,4′-dihydroxy-5-methoxystilbene,cochinchinenin B,4′-hydroxy-4,2′-dimethoxy-dihydrochalcone(a new compound),cochinchinenin A,and loureirin B,respectively.The other might be the original constituent or the metabolite.Conclusion The six constituents absorbed into blood are possible bioactive components of Resina Draconis in vivo.Further research will help clarify the bioactive constituents and mechanisms of Resina Draconis.

Objective To investigate the density of bladder pacemaker cells (ICC) in prostatic hyperplasia with detrusor underactivity (PDU) and the impact of stress environment on its growth. Methods 23 PDU guinea pig models and 26 pigs for control (BPH, n = 6; normal group, n = 20) were developed. After water-filled urodynamic pressure measurement of bladder for three groups , detrusor tissue was made into frozen sections. TheICC numberwere measured with immunofluorescence stainingand observed by confocal microscopy. Further ICC cells in vitro were cultured under stress environment and its counts were analyzed by colony formation assay. Results Decreased density of ICC was observed in PDU group (0.28 ± 0.52)% compared to control [(5 ± 0.36)%,P < 0.05] and the BPH group (5.4% ± 0.42)%, P < 0.05). The ratio of dimer and monomer [(6.13 ± 1.45)%] in PDU group was significantly lower than those of the control [31.55 ± 9.67)%, P < 0.05] and the BPH group [(71.52 ± 8.95)%, P < 0.05]. There is a negative correlation between the density of ICC in vitro and duration of stress. By contrast , there is a positive correlation between the density of ICC and incubation time after the stress is removed. Conclusion Distribution of decreased density of ICC is involved in the pathogenesis of PDU , which may be due to long-term stress environment. Thus eliminating stress environment may help restore ICC density in PDU.

Objective To evaluate the protective action of Phyllanthus orinaria(PU)for immune liver injury in mice.Methods Forty-eight NIH mice were allocated randomly to high-dose(HG)PU group,low-dose(LG)PU group,bifenbate group(BG),normal control group(NCG),and model group(MG).From the first day of establishing the models,20 g ? kg-1 of PU solution was given for gastric perfusion(GP)to HG PU group,10 g ? kg-1 of PU solution for GP to LG PU group,0.15 g?kg-1 of bifenbate solution for GP to BG group,and the equal amount of normal saline to the normal control group and the model group,qd for 12 days.Serum ALT level and visceral parameters of liver and spleen were determined,and the liver pathological feature was also examined.Results The ALT level,parameters of liver and spleen were markedly decreased in BG group(P 0.05).Conclusion PU has a good effect on counteracting the immune live injury in mice by markedly decreasing the activity of serum transaminase and visceral parameters of liver and spleen,and improving the necrosis of hepatic cells.

Objective To explore the predictive value of plasma cell-free DNA (cf-DNA) for prognosis in patients with sepsis. Methods 105 patients with sepsis admitted to department of emergency of the First Hospital of Quanzhou Affiliated to Fujian Medical University from June 2015 to June 2017 were enrolled. Patients were divided into sepsis group (n = 50) and severe sepsis group (n = 55). At the same time, 50 cases of physical examination center in our hospital were randomly selected as the healthy control group. The differences of cf-DNA, procalcitonin (PCT) and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score among the three groups were compared. The correlation between cf-DNA and PCT or APACHEⅡ were analyzed by Bivarite method. Logistic regression was used to analyze the independent predictors of sepsis. The receiver operating characteristic curve (ROC) was made to evaluate cf-DNA, PCT, APACHEⅡ alone or combined ability to predict the prognosis of sepsis. Results The PCT, APACHE Ⅱ and cf-DNA in the sepsis group and severe sepsis group were significantly higher than those in the healthy control group [PCT (μg/L):5.80 (3.28, 8.85), 17.53 (8.40, 29.61) vs. 0.02 (0.01, 0.03); APACHEⅡ: 13.04±3.03, 23.29±8.02 vs. 2.10±1.05;cf-DNA (μg/L): 1 438.0 (1 154.0, 1 576.0), 2 595.0 (2 162.0, 5 198.0) vs. 17.0 (13.0, 20.5); all P﹤0.05], and the indicators in the severe sepsis group were further higher than the sepsis group (all P < 0.05). Correlation analysis showed that cf-DNA was significantly positive correlated with PCT [r = 0.675, 95% confidence interval (95%CI) = 0.575-0.766, P < 0.001] and APACHEⅡ (r = 0.911, 95%CI = 0.874-0.939, P < 0.001). ROC curve analysis showed that the areas under ROC curve (AUC) of PCT, APACHEⅡ, cf-DNA, PCT+APACHEⅡ, cf-DNA+PCT, cf-DNA+APACHEⅡ, cf-DNA+PCT+APACHEⅡ to predict the prognosis of sepsis patients were 0.898, 0.905, 0.961, 0.941, 0.974, 0.976 and 0.982, respectively. It was shown that when predicted alone with PCT, APACHEⅡ and cf-DNA, the AUC of cf-DNA was the largest (0.961), the sensitivity was 100%, and the specificity was 81.43%; the combined prediction of cf-DNA with PCT or APACHEⅡ could further increase AUC, and the combined prediction of cf-DNA, PCT and APACHEⅡhad the highest AUC (0.982), the sensitivity was 94.29%, the specificity was 98.57%. Conclusions cf-DNA, PCT and APACHEⅡ have certain predictive value for the prognosis of sepsis. The value of cf-DNA was the highest when predicted alone, but the predictive ability of cf-DNA combined with PCT and APACHEⅡ was the best.

Objective: To study the epidemiologic and genetic characteristics of coxackievirus A6(CV-A6) strains isolated in Shenyang. Methods: Enterovirus strains positive for neither enterovirus A71 (EV-A71) nor CV-A16 were isolated from Shenyang during 2013 to 2017 to screen for CV-A6 isolates by real-time PCR. The entire sequences of viral genes encoding VP1 of CV-A6 positive samples were amplified and sequenced. The phylogenetic analysis was performed. Results: CV-A6 strains accounted for 27.83% (575/2 066) of the non-EV-A71 and non-CV-A16 enterovirus strains isolated in Shenyang during the years 2013 to 2017. And CV-A6 strains were the predominant enterovirus strains with positive rate of 68.38 % (240/351) in 2015. The CV-A6 isolates from Shenyang during 2013 to 2017 could be classified into the cluster D3a in the phylogenetic tree. Subtype D3a.1 strains circulated during 2013 to 2014 and subtype D3a.2 strains circulated during 2015 to 2017. Conclusions CV-A6 strains were the predominant enterovirus strains among non-EV-A71 and non-CV-A16 enterovirus strains circulated in Shenyang from 2013 to 2017. The CV-A6 isolates from Shenyang during 2013 to 2017 could be classified into the cluster D3a in the phylogenetic tree and subtype D3a.2 strains were evolved from subtype D3a.1 strains.

Ascites ; therapy ; Humans ; Treatment Outcome ; Ultrafiltration ; instrumentation

Objective:To explore the feasibility and safety of the clinical application of the diagnosis and treatment mode combining rapid frozen pathological examination of prostate biopsy tissue with radical prostatectomy.Methods:Suspected prostate cancer patients with PSA>10 ng/ml and PI-RADS score ≥4 in, Northern Jiangsu People's Hospital from April to September 2021 were collected. The included patients underwent mpMRI/TRUS image fusion-guided transperineal prostate targeted biopsy with 16G biopsy needle, 2-3 needles for biopsy, and rapid frozen pathological examination. Robot-assisted laparoscopic radical prostatectomy (RALP) was performed immediately for patients with prostate cancer with rapid freezing pathology. For undiagnosed prostate cancer, 18G biopsy needle for prostate targeted + systematic biopsy were used, 18-22 needles for systematic biopsy, and routine pathological examination. The baseline data, frozen pathological results, perioperative conditions, pathological results and follow-up data of all patients were collected.Results:Eleven patients were included in the study, the mean age of the patients was 69.9(66-73) years, the mean BMI was 22.8(19-26) kg/m 2, the mean PSA was 23.2(14.25-32.00), the mean prostate volume was 45(32-52) ml, mean PSAD 0.54(0.33-0.75). PI-RADS score was 4 in 3 cases and 5 in 8 cases; digital rectal examination was positive in 5 cases. All 11 cases underwent rapid freezing and the pathological results showed that: 9 cases were prostate adenocarcinoma, and RALP was performed immediately. The operation time was 111.5(96-126) min, the intraoperative blood loss was 78.9(55-105) ml, and the postoperative extubation time was 4.3(3.5-5.0) days, postoperative hospital stay 5.8(5.0-6.5) days. Postoperative pathology showed that Gleason score 3+ 4=7 in 1 case, 4+ 3=7 in 3 cases, 8 points in 4 cases, and 10 points in 1 case; 3 cases had positive resection margins, and 1 case had seminal vesicle invasion, the average number of dissected lymph nodes was 10.9 (8.5-14.0), and there was no tumor metastasis. Pathological T staging included 2 cases of T 2b stage, 5 cases of T 2c stage, 1 case of T 3a stage, and 1 case of T 3b stage. Two patients were undiagnosed by rapid freezing pathology, of which one was prostate adenocarcinoma with a Gleason score of 4+ 3=7, and then received RALP; the other one was prostate inflammation. 11 patients were followed up; the postoperative follow-up time was 3-7 months, with an average of 5.2 months. Among the 10 patients who underwent RALP, 8 patients recovered urinary continence 2 weeks after surgery, and all recovered within 2 months after surgery. Three patients with positive surgical margins were given regular androgen deprivation therapy in the second week after surgery. PSA did not drop below 0.1 ng/ml in patients with positive margins and seminal vesicle invasion 3 months after surgery. No complications of Clavien grade Ⅰ or higher occurred after operation and during follow-up. Conclusions:For patients with high suspicion of prostate cancer, rapid frozen pathological examination of prostate biopsy tissue is performed. RALP is performed immediately for patients with prostate cancer. The results show that this diagnosis and treatment model could be safe and feasible.

Objective: To investigate the clinicopathological characteristics and diagnosis of ovarian Brenner tumors. Methods: Forty-seven cases of ovarian Brenner tumors were enrolled from January 2012 to May 2018 at Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Clinical data, imaging examination, histopathological characteristics and immunohistochemical phenotype were analyzed. Results: The age of the patients ranged from 30-73 years and the mean age was 55 years. Thirty-nine patients (83.0%) were postmenopausal. Forty cases (85.1%) of the Brenner tumors were benign, five (10.6%) borderline and two (4.3%) malignant. Usual tumor markers of ovarian carcinoma, including CA199 and CA125 were normal or mild elevated in the 47 cases. Imaging before surgery was not specific to Brenner tumors. Microscopically, benign Brenner tumors were composed of nests of bland, transitional-type cells within a fibromatous stroma. In our 5 cases of borderline Brenner tumors, mildly atypical transitional-type cells were projected into the cyst lumens and lack of stromal invasion. In 2 cases of malignant Brenner tumors, different degrees of nuclear atypial transitional-type cells exhibited stromal invasion. Immunohistochemical stains for CK7, GATA3, p63 and CK5/6 were positive in all cases. Ki-67 was less than 5% in Brenner tumors, and up to 20%-30% in malignant Brenner tumors. Conclusion Brenner tumors are mostly seen in postmenopausal patients and are usually benign. Imaging examination and usual ovarian tumor markers do not provide diagnostic value. Diagnosis and classification of Brenner tumors depend on histopathological evaluation.