Objective To analyze the characteristics of 5 152 cases HPV genotyping in Guangxi province ,which will be benefit for the control of HPV infection and provid experimental evidence for clinical treatment .Methods Statistically analyze the positive detection rate of all the HPV subtypes ,the differences in the positive rate between people of different genders and ages .Results The total positive rate was 37 .46% (1 930/5 152) .The subtypes with the top seven positive rates were HPV16 (5 .90% ) ,HPV52 (5 .36% ) ,HPV58(4 .04% ) ,HPV6(3 .40% ) ,HPV53(2 .66% ) ,HPV11(2 .43% ) ,HPV18(2 .19% ) ,which were mainly high‐risk subtypes .The total positive rate of male patients was 87 .71% ,while female patients was 34 .45% ,the total positive rate of male pa‐tiets was higher than women .For the positive rate of HPV6 ,HPV11 and HPV58 ,male patients were higher than women ,while for HPV52 female patients was higher than men(P<0 .05) .High‐risk HPV6 ,HPV11 ,HPV42 ,HPV43 infection were characterized by the tendency of younger patients ,the differences were statistically significant(P<0 .05) ,the positive infection rate of patients equal or less than 20 years old(75 .51% ) was higher than other age groups .Conclusion HPV infection rates are very high in some re‐gions of Guangxi ,and attention should be paid to male HPV infection .The subtypes with the top seven positive infection rate are mainly high‐risk subtypes .Low‐risk subtypes such as HPV6 ,11 ,42 ,43 are characterized by the tendency of younger patients .The distribution of HPV infection was affected by region ,gender and age .The investigation of HPV subtypes in Guangxi and do HPV screening in different age groups could help the prevention of cervical cancer and understanding HPV infection outcome .

Objective To establish the optimal extracted method for content dete rmination of naringin in Citrus grandis. Methods RP-HPLC was used to determinat e the content of naringin extracted with the above two methods from different ye ar samples of Citrus grandis. Results The average content extracted with ultraso nic extraction was 13.53 %,and the average content extracted with soxhlet extr action was 11.98 %,there being insignificant difference between the two method s. Conclusion The content of naringin extracted with ultrasonic extraction is mo re than that with the soxhlet extraction,which be receipted in Chinese pharmeco pia. And ultrasonic extraction method is more convenient and can save time.

Objective To study the inhibitory effect of curcumin on the proliferation,migration and invasion of non-small cell lung cancer cell A549,and to discuss further if it is closely related to the expression of c-Jun N-terminal kinase (JNK) and relative protein p38.Methods A549 cells were cultured by conventional method,and then treated with different concentration of curcumin (10,20,40,80 μmol · L-1).The proliferation,migration and invasion of A549 cells were measured by real-time cellular analysis (RTCA).The expression levels of JNK,p-JNK,p38 and P-p38 were detected by real-time PCR and Western blotting.Results Curcumin showed an antiproliferation effect against A549 cells with IC50 =40 μmol · L-1,and curcumin exhibited obviously inhibitory effect on the migration and invasion of A549 cells.Additionally,compared with control group,curcumin suppressed the expression of JNK and p38 at the gene level,and significantly inhibited the expression of JNK,P-JNK,p38 and p38 (P＜0.05) at the protein level.Conclusion These results demonstrated that curcumin can inhibit the proliferation,migration and invasion of A549 cells via reducing the level of JNK,p38 phosphorylation,and blocking JNK signal transduction pathway.

Objective: To develop a real-time quantitative PCR (qPCR) assay for detecting Sendai virus gene copy number. Methods: The recombinant plasmid was constructed and a reference used to establish the real-time qPCR method with the TaqMan probe and verified for reproducibility and sensitivity. Results: The linear range of the developed real-time qPCR assay was 1.024 × 10³ ~5 × 10¹⁰ copies/μl, with an R² value of more than 0. 99. The standard recovery of the tested samples was 84.56%~119.48%. Conclusions Compared with traditional hernagglutination assay for particle number detection, this method has higher sensitivity, better repeatability and high quantitative accuracy. It can be used for the detection of particle number of vaccine with sendai virus as the carrier, so as to detect the specific activity of vaccine products.

Objective This study evaluated the effects of health education promoting therapeutic lifestyle changes in a population with dyslipidemia. Methods Patients with dyslipidemia were randomly assigned to one of four groups: the current group (CG) received conventional health guidance, the educational course (EC) group attended six lectures as part of an educational course, the phone call (PC) group received twice-monthly follow-up by telephone,and the comprehensive group(EC+PC)attended both the educational course and received follow-up telephone calls. Total cholesterol (TC), total triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein (HDL-C), and the knowledge, attitude, and behavior (KAP) score for blood lipids were compared within each group and among groups. Results A total of 214 patients were enrolled and completed the study: 62 patients in CG,49 patients in EC,56 patients in PC,and 47 patients in EC+PC.There were significant differences in the EC,PC,and EC+PC groups after the 24-week intervention. For example, pre- and post-intervention values for each group were as follows:EC group:(5.74±0.69)mmol/L and(5.14±0.87)mmol/L for TC,35.22±1.67 and 42.96±5.72 for KAP;PC group:(5.63±0.58)mmol/L and(5.22±1.07)mmol/L for TC, 34.54.±0.97and 39.41±5.03 for KAP;EC+PC group:(5.60±0.48)mmol/L and(4.00±0.79)mmol/L for TC,35.44±1.80 and 45.05±3.19 for KAP, respectively (P<0.05). The CG group showed no significant differences before and after treatment:(5.66±0.54)vs.(5.32±1.28)mmol/L for TC,34.37±0.65 vs.35.28±4.02 for KAP(P>0.05).In a comparison among the four groups,the EC and PC groups showed greater improvements than the CG group.Moreover, the EC+PC group showed statistically significant differences in the results compared with the other three groups (P< 0.05). Conclusion An educational course combined with telephone follow-up calls was more effective than a single intervention in improving blood lipids and enhancing the health awareness of patients with dyslipidemia.This combined health education model not only improves the effectiveness of treatment to some degree,but also plays a role in its supervision and management.Furthermore,it may also assist in the implementation of continuous nursing services in medical institutions.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.

Objective:To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.

Objective To study the effect of lipopolysaccharide(LPS)-induced M1 polarization of macrophages on tumor growth.Methods Female Balb/c mice were randomly divided into control group and experimental group.Renca cells were used to establish subcutaneous tumor model.NaCl(100μl/mice,once every two days)or LPS(100μg/mice,once every two days)was injected to the tumor side when the tumor grew to 50mm3.The M1 polarization level of tumor-associated macrophages was detected by flow cytometry.Mouse tumor cell lines(ML-1,MC38,Renca)were divided into three groups:blank group(complete medium with 50％DMEM basal medium),control group(complete medium with 50％medium supernatant of cultured macrophages)and experimental group(complete medium with 50％medium supernatant of LPS pretreated cultured macrophages).The proliferation of tumor cells was detected by cell counting kit-8(CCK-8).The cell cycle and apoptosis of tumor cells were detected by flow cytometry.Results The proportion of tumor-associated macrophages M1 increased after LPS treatment(P<0.001),thus enhancing its anti-tumor function.LPS-induced M1 polarization of macrophages can significantly inhibit the proliferation of tumor cells(Renca:P=0.023,ML-1:P=0.045).LPS-induced M1 polarization of macrophages blocked G0/G1 phase(MC38:P=0.011,ML-1:P=0.022)or S phase(Renca:P=0.022)of tumor cell cycle,and then cell division was inhibited.LPS-induced M1 polarization of macrophages significantly induced apoptosis of tumor cells(Renca:P=0.04,ML-1:P=0.007).Conclusion LPS can play an anti-tumor role by inducing M1 polarization of macrophages.

To develop a specific, rapid, and convenient immunochromatography assay (ICA) to detect the rabies antibody in clinical sample from immuned dogs by rabies vaccine. Colloidal gold particles labeled with purified rabies virus (CVS11) were used as the detector reagent. The staphylococcal protein A (SPA) and pured rabbit anti-rabies virus IgG were blotted on the test and control regions of nitrocellulose membrane. Then the strip was assembled with sample pad, absorbing pad, and dorsal shield. The assay samples (261 dog's serum) were collected from Wildlife Rabies Disease Diagnostic Laboratories of Ministry of Agriculture in China, Institute of Military Veterinary, Academy of Military Medical Sciences and other six provinces, including rabies virus positive and negative serum. The performance of the strip was compared to fluorescent antibody virus neutralization test. The neutralizing antibody titer could be detected above 0.5 IU. The strip did not change of performance when stored at room temperature for 12 months. It may offer reference of neutralizing antibody titer level after dogs immuned rabies vaccine and determin whether the dogs need to be immuned again.
Animals ; Antibodies, Neutralizing ; analysis ; blood ; Antibodies, Viral ; analysis ; blood ; Dogs ; Gold Colloid ; Immunochromatography ; methods ; Rabies ; prevention & control ; veterinary ; Rabies Vaccines ; immunology ; Rabies virus ; immunology ; Reagent Strips ; Sensitivity and Specificity ; Vaccination

Objective To explore the effects and possible mechanism of histone deacetylase inhibitor SAHA on the interstitial fibrosis induced by diabetes.Methods The SD rats were divided into three groups:control group (Con,n=9),diabetes mellitus (DM) group (n=9) and SAHA treatment group (n=9).The diabetic rat model was established by injecting streptozotocin (STZ) through tail vein.After 8 weeks,the SAHA treatment group rats were fed with a SAHA solution (25 mg· kg-1 · d-1) by gastric gavage.After 16 weeks,all rats were sacrificed to detect relevant biochemical parameters,and observe the changes of pathomorphology in kidney.In addition,immunohistochemistry staining and Western blotting were employed to detect the protein expressions of transforming growth factor-β1 (TGF-β1),Smad2,Smad3,p-Smad2,p-Smad3,Smad7,collagen-Ⅰ and collagen-Ⅲ,respectively.Results Compared with Con group,the levels of blood glucose (BG),urinary trace albumin/urinary creatinine (ACR),triglyceride (TG) and total cholesterol (TC) in the diabetic group were all increased significantly (all P ＜ 0.05),the protein expressions of TGF-β1,p-Smad2,p-Smad3,collagen-Ⅰ and collagen-Ⅲ in kidney were all increased in diabetic group (all P ＜ 0.05),and the expression of Smad7 was significantly reduced (P ＜ 0.05).Compared with DM group,the levels of ACR was reduced,the renal fibrosis was alleviated,the protein expressions of TGF-β1,p-Smad2,p-Smad3,collagen-Ⅰ and collagen-Ⅲ in SAHA group were all decreased (all P ＜ 0.05),and the expression of Smad7 was increased significantly (P ＜ 0.05).Conclusion SAHA may restore the protein level of Smad7 by enhancing protein stability,then promote the moderate transduction of TGF-β1/Smads signaling pathway,which reduce the fibrosis of renal tubules in diabetic rats.