The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×10⁷ U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.
Swine ; Animals ; Cricetinae ; Interferon-gamma/pharmacology* ; Cricetulus ; CHO Cells ; Sincalide ; Recombinant Proteins/pharmacology* ; Antiviral Agents/pharmacology*

The aim of this study was to produce E^rns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified E^rns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-E^rns was constructed based on the gene sequence of BVDV-1 NADL strain. The E^rns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-E^rns into CHO cells. The expression and purification of the E^rns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the E^rns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified E^rns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the E^rns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the E^rns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log₁₀ was induced in rabbits. In this study, purified BVDV E^rns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.
Rabbits ; Animals ; Cricetinae ; Cricetulus ; CHO Cells ; Antibodies, Viral ; Diarrhea Viruses, Bovine Viral/genetics* ; Antibodies, Monoclonal/genetics* ; Diarrhea ; Viral Vaccines/genetics*

Objective:To explore the effect of WeChat-based education and rehabilitation program (WERP) on anxiety, depression, quality of life in patients with non-small cell lung cancer (NSCLC) after radical resection for lung cancer.Methods:From June 2018 to May 2020, convenience sampling was used to select 200 patients with NSCLC who underwent radical resection for lung cancer in the First People's Hospital of Wenling as the research subject. According to the random number table, the patients were divided into the WeChat group and the control group, with 100 cases in each group. The control group was given routine nursing, and the WeChat group received WERP. The scores of Hospital Anxiety and Depression Scale (HADS) and European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (QLQ-C30) were compared between the two groups before and after the intervention.Results:After the intervention, the HADS anxiety and depression scores in the WeChat group were lower than those in the control group, and the QLQ-C30 overall health and function scores were higher than those in the control group, with statistically significant differences ( P<0.05) . Conclusions:WERP is an effective way to relieve anxiety and depression in patients with NSCLC after radical resection for lung cancer and improve their quality of life.

Objective To evaluate the effect of percutaneous coronary intervention (PCI) on anxiety and depression among patients with coronary heart disease (CHD). Methods A total of 600 CHD patients were divided into the stent group (n = 400) and the non-stent group (n = 200) according to stent implantation during coronary angiography, and 400 cases in the stent group were further divided into the intervention group (n = 200) and the non-intervention group (n = 200) according to post-stenting psychological interventions. The anxiety and depression were scored using self-rating anxiety and depression scales 1 day after admission, 1 day after PCI and at discharge from hospital, and the anxiety and depression scores were compared at different time points. Results The mean age, sex ratio, hemoglobin, total cholesterol, triglyceride, high-density lipoprotein cholesteroland low-density lipoprotein cholesterol levels were comparable among the three groups (P > 0.05). There were no significant differences among the three groups in terms of anxiety or depression scores one day after admission (P > 0.05). One day after PCI, the anxiety and depression scores were significantly higher among CHD patients in the intervention and non-intervention groups than in the non-stent group (P < 0.05), and the anxiety (t = 11.21, P < 0.01; t = 9.96, P < 0.01) and depression scores (t = 8.56, P < 0.01; t = 6.73, P < 0.01) were significantly higher in the intervention and non-intervention groups one day after PCI than one day after admission. At discharge from hospital, there were significant differences among the three groups in terms of anxiety and depression scores (P < 0.05), and the anxiety (t = 21.57, P < 0.01; t = 15.77, P < 0.01) and depression scores (t = 24.33, P < 0.01; t = 15.01, P < 0.01) were significantly higher in the intervention and non-intervention groups at discharge from hospital than one day after PCI, while the anxiety and depression scores were significantly lower among CHD patients in the intervention group than in the non-intervention group (P < 0.05). Conclusion The anxiety and depression are aggravated among CHD patients after PCI, and psychological interventions may alleviate the anxiety and depression.

Objectives: High rates of syphilis have been reported worldwide among men who have sex with men (MSM). This study aims to describe the clinical pattern and treatment response of syphilis among human immunodeficiency virus (HIV)-infected MSM in Malaysia. Methods: This is a retrospective study on all HIV-infected MSM with syphilis between 2011 and 2015. Data was collected from case notes in five centres namely Hospital Kuala Lumpur, Hospital Sultanah Bahiyah, Hospital Umum Sarawak, University of Malaya Medical Centre and Hospital Sungai Buloh. Results: A total of 294 HIV seropositive MSM with the median age of 29 years (range 16-66) were confirmed to have syphilis. Nearly half (47.6%) were in the age group of 20-29 years. About a quarter (24.1%) was previously infected with syphilis. Eighty-three patients (28.2%) had other concomitant sexually transmitted infection with genital warts being the most frequently reported (17%). The number of patients with early and late syphilis in our cohort were almost equal. The median pre-treatment non-treponemal antibody titre (VDRL or RPR) for early syphilis (1:64) was significantly higher than for late syphilis (1:8) (p<0.0001). The median CD4 count and the number of patients with CD4 <200/μl in early syphilis were comparable to late syphilis. Nearly four-fifth (78.9%) received benzathine-penicillin only, 5.8% doxycycline, 1.4% Cpenicillin, 1% procaine penicillin, and 12.4% a combination of the above medications. About 44% received treatment and were lost to follow-up. Among those who completed 1 -year follow-up after treatment, 72.3% responded to treatment (serological non-reactive – 18.2%, four-fold drop in titre – 10.9%; serofast – 43.6%), 8.5% failed treatment and 17% had re-infection. Excluding those who were re-infected, lost to follow-up and died, the rates of treatment failure were 12.1% and 8.8% for early and late syphilis respectively (p=0.582) Conclusion: The most common stage of syphilis among MSM with HIV was latent syphilis. Overall, about 8.5% failed treatment at 1-year follow-up.

Objective： ：To explore a novel chimeric antigen receptor (CAR)-T cell treatment to treat Multiple Myeloma (MM) via target B cell maturation antigen (BCMA). Methods： ：A CAR-BCMA molecular was constructed based on mouse originated BCMA scFv, and was packaged into lentiviral vector and transfected into T cells from healthy donors to construct CAR-BCMA-T cells. The BCMApositive cell lines A549-BCMA, A549-BCMAOFP and K562-BCMA were constructed as target cells. Then, the CAR-BCMA-T cells were co-incubated with the constructed target cells and human myeloma U266 cells, and the cytotoxic effects of CAR-BCMA-T cells were evaluated via CCK-8 and FACS. Finally, the CAR-BCMA-T cells originated from MM patients were constructed, and its cytotoxicity against A549-BCMA were examined; in addition, the IFN-γ release level in CAR-BCMA-T cells was evaluated by ELISA and FACS. Results: After 11 days’incubation, the CAR-BCMA-T cells originated from healthy donors amplified 300 times with a positive rate of 43%. The BCMApositive target cell lines were constructed successfully. Under an effector : target ratio of 5:1, the killing rates of CARBCMA-T cells against A549-BCMA, K562-BCMA and U266 were about 80%, 60%, and 80%, respectively, which were significantly higher than those against BCMA negative cells; and the cytotoxicity was related to the BCMA expression level in target cells. What’ s more, at the effector : target ratio of 20:1, the CAR-BCMA-T cells originated from MM patients were demonstrated to exhibit a killing rate of more than 95% againstA549-BCMApositive cells, and produced large amount of IFN-γ. Conclusion: CAR-BCMA-T cells originated from both healthy and MM donors were successfully constructed, and they can effectively and specifically kill BCMA positive tumor cells.

Objective To investigate the effect of Shenxiong glucose injection on platelet reactivity during aspirin treatment in patients with acute ischemic stroke.Methods A total of 263 patients with acute ischemic stroke admitted to 12 hospitals from January 2014 to December 2016 were enrolled prospectively.They were randomly divided into aspirin group and aspirin + Shenxiong glucose injection group.The changes of platelet maximum aggregation rate induced by 4 platelet aggregating agents (arachidonic acid,adenosine diphosphate,collagen and platelet activating factor) were detected before and after the treatment.Results There were no significant differences in the demographic data and baseline clinical characteristics between the aspirin group (n =132) and the Shenxiong glucose injection + aspirin group (n =131).At baseline,the maximum aggregation rate of platelet induced by arachidonic acid and platelet activating factor in Shenxiong glucose injection + aspirin group was significantly higher than that in the aspirin group (all P ＜0.05).On the 6th day after treatment,the maximum aggregation rate of platelets induced by the 4 aggregating agents in the Shenxiong glucose injection + aspirin group was significantly lower than that in the aspirin group (all P ＜ 0.001).Conclusion Shenxiong glucose injection had a significant inhibitory effect on platelet reactivity during aspkin treatment in patients with acute ischemic stroke.

Osteopontin (OPN) is a widely distributed secretory protein and is involved in integrin-mediated cell signal transduction. It also plays an important role in the development, progression, and metastasis of hepatocellular carcinoma (HCC). OPN plays a pivotal role in biomineralization and innate immunity, participates in tumor growth and angiogenesis by interacting with integrin, and induces the recurrence and metastasis of HCC via multiple signal transduction. Combined measurement of OPN and other tumor biomarkers can improve the sensitivity of early diagnosis of HCC. OPN inhibitors may serve as a new target for the treatment of HCC and can improve the sensitivity of chemotherapeutic drugs. This article summarizes the recent studies on OPN and introduces the biological function of OPN and its role in the progression and diagnosis and treatment of HCC.

Xanthogranulomatous cholecystitis (XGC) is a rare type of chronic cholecystitis characterized by severe proliferative fibrosis with infihration of macrophages and foamy cells in the gallbladder wall.Since XGC and gallbladder carcinoma have similar clinical manifestations and radiological features,XGC is often misdiagnosed as gallbladder carcinoma in clinical practice,which leads to unnecessary extensive surgical resection and has an adverse effect on patients.At present,the preoperative diagnosis of XGC is still based on imaging results (ultrasound,computed tomography,and magnetic resonance imaging),and a definite diagnosis of this disease relies on intraoperative frozen biopsy or postoperative pathological examination.Meanwhile,XGC should be differentiated from gallbladder adenomyomatosis,gallbladder carcinoma,and gallbladder actinomycosis.Laparotomy or laparoscopic cholecystectomy is the major method for the treatment of XGC,but laparoscopic cholecystectomy is associated with a longer time of operation,more complications,and a higher rate of conversion to laparotomy.Therefore,surgeons are facing difficulties in preoperative diagnosis and intraoperative decision-making process of XGC.

Colistin is a kind of old cationic drug,which can interfere bacterial cell membrane,thus to cause bacterial death.It is mainly used for the treatment of infections caused by Gram-negative bacteria,and its antimicrobial activity against multidrug-resistant Gram-negative bacteria is also very significant.At present,colistin is widely used in veterinary medicine.This article aims to review colistin in chemical,pharmacological,and pharmacokinetic studies,and also summarizes the application and resistance of this drug,which will provide reference for the reasonable selection and use of this drug in animals.