This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

Objective:To understand the monkeypox knowledge awareness, risk perception and vaccination intention in men who have sex with men (MSM) in five cities in northeast China.Methods:A cross-sectional study was conducted by using electronic questionnaire in MSM selected by convenience sampling in five cities in northeast China (Shenyang, Panjin, Changchun, Harbin and Jiamusi) from June 28 to July 8, 2023 by local centers for disease control and prevention and MSM communities. The sample size was estimated to be 220. Information about their demographics, monkeypox-related knowledge awareness, perceived concern about epidemic risk perception, and monkeypox vaccination intention were collected. Logistic regression model was used to analyze related factors for MSM's monkeypox vaccination intention.Results:In 355 MSM, 63.9% (227/355) had monkeypox vaccination intentions, and 55.5% (197/355) had high awareness of monkeypox related knowledge with a mean knowledge awareness score of 3.7±1.5. MSM with education level of high-school and above (a OR=1.93, 95% CI:1.01-3.69), higher knowledge awareness score (a OR=1.19, 95% CI:1.02-1.40) and higher risk perception of monkeypox infection (a OR=1.82, 95% CI:1.15-2.88), were more willing to receive monkeypox vaccination. The main reasons for willingness to receive monkeypox vaccine were preventing monkeypox (86.3%, 196/227) and worrying about appearance being affected (62.1%, 141/227). The main reasons for unwillingness for the vaccination included concerns about vaccine safety (53.1%, 68/128), clinical progression of AIDS being affected (46.1%, 59/128) and efficacy of antiretroviral therapy being affected (44.5%, 57/128). Conclusions:The levels of knowledge awareness and vaccine intentions still need to be improved among MSM in five cities of northeast China. It is necessary to improve the awareness of monkeypox and intention of monkeypox vaccination, promote protected sex behavior and self-assessment of infection risk, reduce vaccine hesitancy and increase monkeypox vaccination intention in MSM in 5 cities in northeast China.

Cholestatic liver disease is a common disease that causes bile flow dysfunction due to various reasons. The etiology of cholestatic liver disease is complexed, and therapeutic drugs are extremely limited. To date, ursodeoxycholic acid is the only FDA-approved drug for treating primary biliary cirrhosis, whereas its efficacy is limited to early stage of the disease, therefore novel drugs are urgently needed. Nuclear receptors become therapeutic hotspot target in cholestasis since these receptors play a key role in regulating bile acid homeostasis. Peroxisome proliferator-activated receptor (PPAR) is an important nuclear receptor involved in regulating multiple mechanisms of cholestasis in vivo. It can improve intrahepatic cholestasis by inhibiting bile acid synthesis, reducing bile acid toxicity, affecting the expression of bile acid metabolic enzymes and transporters, and can play an anti-inflammatory, anti-oxidation and anti-fibrosis role. A number of studies have shown that PPAR agonists represented by fibrates alone or in combination can improve liver function indexes, inflammatory factors and fibrosis markers in patients with cholestasis. This review analyzes and summarizes the lastest advances in the molecular mechanism of PPAR as a therapeutic target for cholestasis and drug treatment in development or have been used in clinical.

OBJECTIVE To monitor the occurrence of tenofovir disoproxil fumarate （TDF）-induced kidney injury and investigate the risk factors， and provide reference for rational use of TDF in clinic. METHODS The information of inpatients with hepatitis B was collected by China Hospital Pharmacovigilance System （CHPS） from the Second People’s Hospital of Lanzhou during Jan. 1st， 2019 to Dec. 31st 2021. The search criteria were set according to kidney injury criteria， and suspected TDF- induced kidney injury cases were actively monitored； then the clinical pharmacist confirmed the positive patients with TDF-induced kidney injury one by one and calculated the incidence of TDF-induced renal injury； the risk factors for TDF-induced kidney injury in real world were explored by collecting and analyzing the correlation of basic data of patients， main indexes of liver and kidney function， complications and combined use of drugs with TDF-induced renal indexes. RESULTS Totally 1 226 inpatients with hepatitis B using TDF were included. Through active monitoring of CHPS， 160 suspected patients with TDF-induced kidney injury were found， and 64 positive patients were finally confirmed manually. The incidence of TDF-induced kidney injury was 5.22%. Compared with pre-medication， the levels of serum creatinine and cystatin C， the proportion of patients with urinary protein 2+ and above were increased significantly after medication （P＜0.001）， glomerular filtration rate and blood phosphorus level were reduced significantly （P＜0.001） and other indicators had no statistical difference. Treatment time for more than 36 months， disease progresses to decompensated cirrhosis， and concomitant use of more than 10 kinds of drugs were significantly correlated with TDF- related kidney injury （P＜0.05 or P＜0.012 5）. CONCLUSIONS The active monitoring scheme of TDF-induced kidney injury established by CHPS has the characteristics of time-saving， labor-saving and high efficiency； based on real-world evidence， it is imperative to strengthen monitoring kidney function of patients when using TDF， especially when the patient has been on medication for a long time， in decompensated cirrhosis and combination of multiple drugs， and thus， we can identify earlier and avoid adverse effects in high-risk patientseffectively.

Objective To investigate the changes and clinical correlation of IL-4 in patients with HBV infection.Methods Sixty cases with chronic asymptomatic HBV carriers,60 cases with chronic hepatitis B,60 cases with liver cirrhosis,60 cases with hepato-cellular carcinoma and 50 healthy controls were collected for serum.ELISA was used for detection of cytokine IL-4 levels;The a-mount of HBV DNA was measured by fluorescence quantitative PCR;Liver function was tested by automatic biochemical analyzer. Results Compared with the healthy controls[(1.64±0.17)ng/mL],IL-4 levels of patients with CHB,LC and HCC were signifi-cantly increased[(4.18±0.48),(4.71±0.42),(3.62±0.31)ng/mL,P <0.05].LC group have the highest IL-4 levels,while ASC was the lowest.Compared with the ASC group,IL-4 levels of patients with CHB,LC and HCC were significantly increased(P <0.05).Compared with the HCC group,IL-4 levels of patients with LC were significantly increased(P <0.05).IL-4 level was posi-tively correlated with TBIL levels in LC patients(r=0.529,P <0.01),while the IL-4 level was positively correlated with ALT and TBIL level in HCC patients(r=0.263,0.323,P <0.05).Conclusion IL-4 may play an important role in chronic HBV infection, the levels of IL-4 can be used as an important indicator to assess the severity of chronic hepatitis B.

Objective To establish the quality standard of Jiangzhi Fugan Capules. Methods TLC was used to the qualitative identification of Paeoniae Radix Rubra and Salviae Miltiorrhizae Radix et Rhizoma. The contents of Paeoniflorin and Tanshinone ⅡA were determined by HPLC. The HPLC separation was performed on Phenomenex C18 column (4.6 mm×250 mm, 5 μm), with methanol-0.1% phsophonic acid (31∶69, 75∶25) as mobile phase, and the flow rate was 1.0 mL/min. Results The results of TLC showed that relevant spots were clear without interference against the negative sample. The calibration curves for Paeoniflorin and Tanshinone ⅡA were found to be liner within the range of 0.448-4.48μg, 0.057 6-0.576μg, respectively. The correlation coefficients were 0.999 4 and 0.999 5, respectively. The average recoveries were 99.93%and 99.75%, with RSD of 1.98%(n=9) and 1.70%(n=9), respectively. Conclusion The method is accurate, reliable, stable, rapid, and reproducible, and can be used for the quality control and evaluation of Jiangzhi Fugan Capules.

Objective To investigate the value and clinical significance of CD4+CD25+ regulatory T cells (Treg) ,inteleukin‐18 , interferon‐γand transforming growth factor‐βin patients with hepatitis B virus (HBV) infection .Methods A total of 175 patients with HBV infection were divided into chronic type B hepatitis (CHB) group and chronic asympotomatic HBV carrier (ASC) group , which were further divided into hepatitis B e antigen (HBeAg) positive and negative groups .CD4+ CD25+ Treg ,cytokines levels and liver function were measured .Healthy subjects were enrolled into control group .Results Proportion of CD4+CD25+ Treg were without significant difference among healthy controls ,HBeAg(+ ) and HBeAg(-) ASC groups ,and HBeAg(+ ) and HBeAg(-) CHB groups (P>0 .05) .Compared with control group ,cytokines levels were significant higer in CHB and ASC group (P<0 .05) . CD4+CD25+ Treg level was significant positive correlation to alanine aminotransferase and ratio of aspartate aminotransferase to plateles in HBeAg(+ )ASC group (P<0 .05) .Conclusion CD4+CD25 + Treg and related cytokines could play important roles in the course of CHB ,while CD4+CD25+ Treg expressing might be correlated with inflammatory degree of hepatitis .

Objective To establish the quality standard for Jinyin Lidan Oral Liquid. Methods Artemisiae Scopariae Herba, Taraxaci Herba, Aurantii Fructus and Schisandrae Chinensis Fructus in Jinyin Lidan Oral Liquid were identified by TLC. The contents of naringin and neohesperidin were determined by HPLC. The HPLC separation was performed on Phenomenex C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of methanol-0.1% phsophonic acid solution and gradient elution. The UV detection wavelength was 254 nm;flow rate was 1.0 mL/min;column temperature was 25 ℃. Results The results of TLC showed that relevant spots were clear, with strong specificity, without interference of negative sample. The calibration curves for naringin and neohesperidin were in good linearity in the range of 0.56-5.60μg (r=0.999 9) and 0.38-3.80μg (r=0.999 8), respectively. The average recoveries were 99.86%and 98.95%with RSD of 2.04%and 2.45%, respectively. Conclusion The qualitative and quantitative determination method is simple, accurate and reliable, with good stability and reproducibility, and can be used for the quality control of Jinyin Lidan Oral Liquid.

Objective To optimize the formative technology for Zhuqi Fugan Granules. Methods By using orthogonal experiment design, and setting parameters of granules as determination standards, concrete density, excipient types and dosage, and consentration of wetting agent were optimized to determine the optimal conditions. Results The optimal forming technology was as follows:The optimal ratio of starch to micrite cellulose was 3∶2;the optimal density of extration was 1.20;the ratio of concrete to excipient was 1∶1;the optimal concentration of wetting agent was 80% of ethyl alcohol. Zhuqi Fugan Granule enjoyed good mobility and with a critical relative humidity of 67%. Conclusion Parameters of Zhuqi Fugan Granules prepared by this method are satisfying, with stable properties and low moisture absorbtion. This preparation technology is simple and feasible.