A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.

Objective To evaluate the efficacy and safety of continuous infiltration of Lidocaine in joint replacement surgery.Methods From May 2013 to October 2013,patients scheduled to undergo total hip or total knee arthroplasty were randomized into two groups:the local infiltration analgesia group received continuous infiltration of Lidocaine while the other group received PCIA filled with fentanyl after surgery.The equipments were removed 48 hours after the surgery.The visual analogue scale score at five points (immediate postoperative,24 hours after the surgery while resting,24 hours after the surgery while exercising,48 hours after the surgery while resting,48 hours after the surgery while exercising) were recorded and if the score was higher than 4,analgesics wound be given to the patients.The vital sign,blood test,ECG,the circumference of lower limbs,the range of motion of the joint,the rehabilitation and complications were also recorded.Results 42 patients were included,11 male and 31 female.There were 25 patients and 36 total knee arthroplasties.17 patients underwent a total of 19 hip arthroplasties.The demographic data,the diagnosis and the surgery type were comparable between two groups.The VAS scores at immediate postoperative,24 hours and 48 hours after the surgery while resting,the average dose of analgesics and the complication rate were statistically different be tween two groups.However,the VAS scores at 24 hours after the surgery and 48 hours after the surgery while exercising,the change of the circumferences of the lower limb,the range of motion of the joint,the rehabilitation satisfactory rate and length of stay were comparable between two groups,while no significant difference was observed.One patient in the PCIA group suffered from incision fat liquefaction and healed after three weeks.There were no infection and subcutaneous hydrocele in both groups.Conclusion Continuous infiltration of Lidocaine is more effective than the PCIA while resting after joint replacement surgery and no inferior to it while exercising.It can decrease the amount of Opioid.

Objective To evaluate diagnostic value of plasma D-dimer (D-D),fibrinogen (Fg) and Ddimer/fibrinogen (D/F) ratio in acute lower extremity deep vein thrombosis (DVT) after artificial joint replacement.Methods From August 2009 to December 2011,273 patients who had undergone total hip arthroplasty (THA) or total knee arthroplasty (TKA) underwent venography of lower extremity deep vein at 35 days postoperatively.According to results of angiography,all patients were divided into DVT group including 66 patients (15 males and 51 females,median age 66 years) and non-DVT group including 207 patients (70 males and 137 females,median age 63 years).Plasma levels of D-D and Fg were measured by immunoturbidimetry and solidification methods respectively preoperatively and at 3-5 days postoperatively.The differences of plasma levels of D-D and Fg and D/F ratio between the two groups were analyzed with statistical method.Furthermore,diagnostic value of postoperative plasma D-D and D/F ratio in acute DVT was assessed using a receiver operating characteristic (ROC) curve.Results There was no statistical significance in preoperative plasma levels of D-D and Fg,and postoperative plasma levels of Fg between two groups.However,the postoperative plasma levels of D-D and D/F ratio were higher in the DVT group than those in the nonDVT group,and there was significant difference between two groups.In addition,the ROC curve analysis showed that the area under curve of postoperative plasma D-D and D/F ratio was 0.688 and 0.684,respectively,but this difference did not reach statistical significance.Conclusion Plasma Fg may be meaningless for the diagnosis of acute DVT after artificial joint replacement.Although plasma D-D and D/F ratio may be useful for diagnosing acute DVT,the diagnostic accuracy is low.

Objective To explore the risk factors of sepsis complicated with hepatic insufficiency in elder patients.Methods With retrospective case-control study method,according to whether or not complicated with hepatic insufficiency,a total of 384 elder sepsis patients were divided into case group (91 patients) and control group(293 patients),then single factor analysis and multiple factors Logistic regression methods were used to screen the risk factors.Results Single factor analysis identified 6 kinds of significant variables,including age(P =0.000),history of alcohol(P =0.006),chronic diseases(P =0.001),arterial blood lactate level (P =0.035),mechanical ventilation (P =0.009) and acute physiology and chronic health evaluation Ⅲ score (APACHE-Ⅲ) scores(P =0.004).Multiple factors Logistic regression analysis showed that age,history of alcohol,chronic diseases,arterial blood lactate level,mechanical ventilation,APACHE-Ⅲ scores were correlated with sepsis complicated hepatic insuffciency in elder patients (P ＜ 0.01 or ＜ 0.05).Conclusion The risk factors which closely relate to sepsis complicated with hepatic insufficiency in elder patients are age,history of alcohol,chronic diseases,arterial blood lactate level,mechanical ventilation,APACHE-Ⅲ scores.

In order to investigate the anti-tumor angiogenesis activity with a recombinant Ag43/FGFR1 chimeric protein (AF) vaccine in a mouse H22 hepatoma model, tumor volume and survival rate of the mice were studied at a 3-day interval. Microvessel density (MVD) was detected by immunohistochemistry. The endothelial deposition of autoantibodies within tumor tissues was examined by immunofluorescent staining, and anti-FGFR1 antibody-producing B cells (APBCs) were tested by enzyme-linked immunospot (ELISPOT) assay. Compared with the three control groups, the tumor volume was significantly decreased and the survival time was significantly prolonged in AF-immunized group (P<0.05). The number of APBCs in AF-immunized mice (129.6+/-10.9) was more than in controls [6.2+/-1.1 (FGFR1), 6.0+/-1.2 (Ag43) and 5.2+/-1.4 (NS), P<0.01]. Moreover, the endothelial deposition of autoantibodies was found in tumor tissues from AF-immunized mice, but not in control groups. MVD in AF-immunized group was significantly lower than in FGFR1-immunized group, Ag43-immunized group and NS group (10.3+/-3.1 vs 39.4+/-8.6 vs 42.3+/-9.8 and 43.6+/-10.6, P<0.01). These findings demonstrated that the AF protein vaccine effectively inhibited tumor angiogenesis and growth via production of autoantibodies against self-FGFR1.

Objective To evaluate the expression and clinical significance of human papillomavirus(HPV)L1 caspid protein in cytologic specimens of the cervix.Methods The HPV L1 caspid protein in cytologic specimens of the cervix was detected by using immunochemistry in 309 women treated in Shenzhen Hospital of Peking University from Jan.2008 to May 2009.According to histological biopsy,of 309 liquid-based cytologic smears,there were 33 cases of normal cervixes or chronic cervititis,168 cases of grade I of cervical intraepithelial neoplasia(CIN I),84 cases of grade Ⅱ/Ⅲof CIN,and 24 cases of squamous cell carcinomas(SCC).Results The positive expression rate of HPV L1 caspid protein was 27.3%,66.7%,25.0%,and 0 in the normal cervixes or chronic cervititis,CIN I and CIN Ⅱ/Ⅲ,and SCC respectively.There was significant difference between CIN I or SCC and the normal cervixes or chronic cervititis,between CIN I with CIN Ⅱ/Ⅲ,and between CIN I or CIN Ⅱ/m with SCC(all P<0.0 1).The positive expression rate of HPV L1 caspid protein was reduced with the lesion progression from CIN I to CINⅡ/Ⅲ to SCC.309 cases were divided into two groups by 30 and 40 years of age,and there was no significant difference in the HPV L1 caspid protein expression between different age groups(P>0.05).The positive expression rate of HPV L1 caspid protein in the over-1000 RLU/PC viral load group was significantly higher than in the under-1000 RLU/PC viral load group(P<0.05).The sensitivity,specificity,positive predictive value,negative predictive value of HPV L1 caspid protein for detecting high-grade cervical lesions were 80.6%,60.2%,52.1% and 85.2%,respectively.Conclusion The positive rate of HPV L1 caspid protein is decreased with the lesion progression from CIN I to CIN Ⅱ/Ⅲto SCC.HPV L1 caspid protein among HPV-positive women can be a helpful molecular biomarker in risk assessment and prognostic prediction.

Objective :To study diagnostic value of interleukin (IL ) and C reactive protein (CRP ) for aged patients with cardiovascular emergencies .Methods : A total of 189 patients with cardiovascular emergencies ,who were main—ly presented as acute chest tightness ,chest pain and dyspnea and treated in our hospital from Jan 2012 to Nov 2016 , were selected as cardiovascular emergency group .Another 150 cases undergoing physical examination simultaneously were selected as healthy control group .Serum levels of inflammatory factors were measured and compared between two groups.Results : Compared with healthy control group ,there were significant rise in serum levels of IL—1 ,IL—2 ,IL—6 ,IL—10 ,CRP ,homocysteine (Hcy) ,N terminal pro brain natriuretic peptide (NT—proBNP) ,malonyl diade—hyde (MDA) ,reduced form of nicotinamide—adenine dinucleotide phosphate (NADPH ) oxidase subunit p22 ,p47 , p67 and gp91 ,tumor necrosis factor (TNF)—α ,endothelin (ET)—1 and its receptor ,cystatin C (CysC) and cardiac troponin T (cTnT) ,and significant reductions in levels of superoxide dismutase (SOD) and glutathione peroxidase (GSHPX) in cardiovascular emergency group , P＝0.001 all.Conclusion : Serum levels of IL and CRP significantly rise in aged patients with cardiovascular emergencies ,and peroxidase is involved in the process .

In order to investigate the anti-tumor angiogenesis activity with a recombinant Ag43/FGFR1 chimeric protein(AF)vaccine in a mouse H22 hepatoma model,tumor volume and survival rate of the mice were studied at a 3-day interval,Microvessel density(MVD)was detected by immunohistochemistry.The endothelial deposition of autoantibodies within tumor tissues was examined by immunofluorescent staining,and anti-FGFR1 antibody-producing B cells(APBCs)were tested by enzyme-linked immunospot(ELISPOT)assay.Compared with the three control groups,the tumor volume was significantly decreased and the survival time was significantly prolonged in AF-immunized group(P<0.05).The number of APBCs in AF-immunized mice(129.6±10.9)was more than in controls[6.2±1.1(FGFR1),6.0±1.2(Ag43)and 5.2±1.4(NS),P<0.01].Moreover,the endothelial deposition of autoantibodies was found in tumor tissues from AF-immunized mice,but not in control groups.MVD in AF-immunized group was significantly lower than in FGFR1-immunized group,Ag43-immunized group and NS group(10.3±3.1 vs 39.4±8.6 vs 42.3±9.8 and 43.6±10.6,P<0.01).These findings demonstrated that the AF protein vaccine effectively inhibited tumor angiogenesis and growth via production of autoantibodies against self-FGFR1.

ObjectiveTo investigate the clinical efficacy of Tenghuang Jiangu tablets （THJGT） combined with oral non-steroidal anti-inflammatory drugs （NSAIDs） in the treatment of osteoarthritis of the knee and its applicable stage based on real-world data， and provide a basis for the rational clinical use of THJGT. MethodA total of 218 cases treated with THJGT combined with oral NSAIDs included in the "THJGT for knee osteoarthritis case registry" from September 2019 to January 2021 were selected as the observation group， and 126 cases treated with oral NSAIDs alone as the control group （CG）. The data of gender， age， body mass index， Kellgren-Lawrence grading scale （K-L scale） score， visual analogue score （VAS score）， Western Ontario and McMaster Universities Osteoarthritis Index （WOMAC） score， swelling grade， joint fear of cold score， back pain and weakness score， and occurrence of adverse events/reactions of the patients in both groups were used for the evaluation of efficacy with full analysis set. The propensity score matching method was used to exclude the influence of confounding factors between groups， and the sub-data sets were established， with which the repeated measures analysis of variance （ANOVA） was carried out to evaluate the efficacy. Visit points were at registration， 4 weeks and 8 weeks after registration. The data were statistically analyzed in Excel 2019 and SPSS 23.0. ResultThe proportion of females in the observation group was 66.06% （144/218）， which was higher than that （58.73%， 74/126） in the control group （χ²=1.846）. The average age in the observation group was （61.12±7.01） years， which was higher than that ［（59.38±5.99） years］ in the control group （W=19 918.50， P<0.05）. The remission rate in the observation group was 98.17% （214/218）. In the observation group， the proportions of the patients at K-L grades Ⅱ and Ⅲ were 64.22% （144/218） and 25.23% （55/218）， respectively. The effect analysis of the whole data set for enrollment and treatment for 8 weeks showed that the VAS score of the experimental group decreased by （3.27±1.24） points on average， which was better than that of the control group [（2.75±1.20）， W=34 179.00， P<0.05]. The average WOMAC score decreased （23.43±11.46） points， which was better than that of the control group [（16.71±8.86）， W=32 387.00， P<0.05]. The average swelling grade decreased （0.63±0.64）， which was better than the control group [（0.33±0.59）， W=33 847.50， P<0.05]. The average score of joint chills decreased （1.90±1.84）， points， which was better than that of control group [（1.40±1.28）， W=35 165.00， P<0.05]. The average lumbar acid fatigue score decreased by （2.02±1.64） points, which was better than that of the control group [（1.10±1.28）， W=32 986.50， P<0.05]. Efficacy analysis of subdata sets for enrollment， 4 weeks of medication and 8 weeks of medication showed that VAS scores of both groups showed a downward trend after treatment， and the improvement of experimental group was more significant than that of control group at 4 weeks， with statistical significance （P<0.05）. After treatment， the total WOMAC score of both groups showed a downward trend， and the improvement of experimental groups was more significant at 4 weeks and 8 weeks （P<0.05）. After treatment， swelling， cold fear grade and lumbar acid fatigue score of both groups showed a decreasing trend,， and the improvement of experimental group was more significant at 8 weeks （P<0.05）. The therapeutic effect analysis of patients in the attack stage and remission stage of the experimental group showed that the total WOMAC score of the two groups showed a downward trend after treatment， and the trend was basically the same， and there was no statistical difference between the two groups at enrollment， 4 weeks after treatment， and 8 weeks after treatment （t=1.675， t=2.068， t=2.364）. The total WOMAC score of the patients in remission stage in the experimental group with K-L grading between grade 0 and grade Ⅲ had statistical significance at 4 weeks after treatment compared with the time of entry （P<0.05， P<0.01）. Group of adverse event rate was 4.13% （9/218）， lower than the control group 10.32% （13/126） （χ²= 5.109， P<0.05）. ConclusionThe population receiving THJGT combined with oral NSAIDs is mostly female， old， in remission， and with K-L grades Ⅱ and Ⅲ. THJGT can enhance the anti-inflammatory and analgesic effects of oral NSAIDs and keep the drug effect in improving joint function and alleviating fear of cold， swelling， and back pain and weakness. The drug combination can be applied to patients in both attack and remission， and the clinical application should take patient's disease stage and degree of osteoarthritis into account. Furthermore， the combination has the potential to reduce the incidence of adverse events caused by NSAIDs.