Objective To investigate the relationship between hepatitis B virus (HBV) infection and cytokeratin 18 (CK18) phosphorylation.Methods Liver tissues were taken from 21 chronic hepatitis B (CHB) patients by liver biopsy and 14 healthy controls were collected.Immunofluorescence double staining and Western blotting were used to detect CK18 pSer33 and CK18 pSer52 phosphorylation.HepG2 cell line was transfected with either 1.3 mer HBV vector or empty control vector.CK18 pSer33 or CK18 pSer52 phosphorylation were tested using immunofluorescence double staining and Western blotting.Total mRNA was extracted from the cells.The expressions of cell division cycle 2 (cdc2) and protein kinase Cε (pKCε) were analyzed by realtime polymerase chain reaction relative quantification.Statistical analyzee were performed by using two-independent samples t test.Results Compared to healthy controls,phosphorylation of CK18 pSer33 in HBsAg stained hepatocytes was significantly higher in CHB patients (t=6.618,P=0.000).However,no difference was found in phosphorylation of pSer52 (t=2.429,P=0.051).Phosphorylation of CK18 pSer33 and expression of cdc2 mRNA in HBV transfected HepG2 were significantly higher in empty vector transfected HepG2 (t=5.365,P=0.006),while no difference was found in phosphorylation of pSer52 and expression of pKCε mRNA (t=1.098,P=0.334).Conclusion HBV infection is significantly associated with phosphorylation of CK18 pSer33.

Objective Objective In order to better keep up with the development of transplantation immunobiology.we established and compared two types of mouse heterotopic heart transplantation,and hope to help further organ transplantation studies.Methods According to the surgical procedures of Ono's type and Chen's type of mouse model of heterotopic heart transplantation with some modification,we performed celiac and cervical heteropotic heart transplantation between iso-strains and hetero-strains,and compared the operation suecess rate,operation time,allografi survival time,and histopathology of those establishment methods.Results The success rates of mouse celiac and cervical heterotopic heart transplantation were 86.7% and 83.3%,respectively,with a non-significant difference(P>0.05) between the two methods of operation regarding the total operation time,survival time of the allografts,and histopathological findings.Conclusions Based on the mastery of microsurgical techniques,the two models of heterotopic mouse heart transplantation can be established equally,and either of them can be considered depending on the particular requirements of studies.

Objective To study the effects of self-made Chinese sherbal gargle on AIDS related oral lesion.Methods 353 hospitalized AIDS patients from June 2007 to December 2009 were divided randomly into the experimental group(179 patients)and the control group(174 patients).ALL the patients were treated with systemic anti-viral therapy while the experimental group was mouthwashed by self-made herbal gargle and the control group with normal saline solution.The incidence of new oral lesion and the changes of the originM lesion were observed.Results The incidence of new oral lesion in the experimental group was obviously lower than that of the control group.The cure rate and effective rate of original oral lesion were much higher than the control group.Conclusions Self-made herbal gargle shows good effect in preventing and treating the AIDS related oral lesion.

Objective To reconstruct the initiative procedure of HIV-1 reverse transcription in vitro and establish a methodology of assessing activity of HIV-1 reverse transcriptase (RT) with real-time PCR Methods The tRNALys-3 gene was amplified from genome in healthy individuals through polymerase chain reaction (PCR), and then T7 transcription promoter was added in 5'-terminal of the tRNALys-3. The tRNA[Lys-3 cRNA product was obtained by applying T7 RNA polymerase through a transcription reaction. The 5'-LTR-PBS DNA was also obtained by transcription reaction from the HIV-1 infectious clone and inserted into pGEM-T easy vectors. 5'-LTR-PBS cRNA was obtained by applying SP6 RNA polymerase whose combining site was located in pGEM-T easy vectors. Then the two RNA samples was catalyzed by two kinds of standard reverse transcriptases (SuperScript Ⅲ and HIV-1 standard reverse transcriptase, respectively) and the cDNA was synthesised. The relative activity of RT was determined with the real-time PCR. Results The tRNALys-3 primer and the SP6-5'-LTR-PBS RNA were procured accurately, whose length were 93bp and 872 bp, respectively. After the following serial dilution of Super Script Ⅲ and HIV-1 standard reverse transeriptase:1 : 10, 1: 100, 1:1 000, 1:10 000, each step of reverse transcription process worked successfully. Real-time PCR results showed that Ct values of the two groups were 13.9, 18. 3, 20. 9, 24. 9 and 20. 4, 25. 5, 28. 7, 32. 5 respectively. Conclusion A novel real-time PCR method is developed to assay the RT activity directly through reconstructing the initiation of HIV reproduction, which may be helpful for clinical management, screening of new antiretroviral drugs, and drug resistance test.

Drugs, Investigational ; Humans ; Lymphoma ; drug therapy ; United States

Objective To establish a mini-pool nucleic acid testing (NAT) assay using multiplex RT-nested PCR for the detection of HIV RNA, and apply it in screening for acute HIV infection among MSM. Methods Frozen EDTA plasma samples collected between Oct. 2008 and Mar. 2009 from 3 HIV infectors during window-period, a total of 30 HIV chronically infected individuals and 97 healthy subjects were used to develop the NAT assay. Plasma samples from 10 cases were pooled into one tube and centrifuged at high speed for the collection of viruses. HIV RNA was extracted. Two pairs of primers were designed according to two conserved regions of HIV RNA ( HXB2 nt 5783-nt 6228 and nt 1235-nt 2012).Multiplex RT-PCR and nested PCR were performed. Individual NAT-reactive samples were confirmed by commercially available NAT assays. The sensitivity and performance efficacy were also evaluated. The assay was then applied to 1 005 plasma specimens from MSM with negative or uncertain HIV antibody test results.These were collected in the same period as the other samples. Results ( 1 ) Two fragments of HIV were amplified successfully with the low detection limit of 162 copies/ml plasma; (2) Results of the mini-pool HIV NAT validation with samples from 3 HIV infectors during window-period were consistent with the expected values; (3) All 30 plasma samples from MSM with positive HIV antibody, which were tested by multiplex RT nested PCR, were found to be HIV RNA positive; (4) One out of 1 005 plasma samples was found to be HIV RNA positive, for this case acute infection was followed-up and sero-conversion was found. Conclusion Mini-pool NAT has good sensitivity, and may be applied to screening HIV RNA among MSM during window-period.

reproducible, and may cover the major circulating strains in China.

Objective To evaluate the repeatability of HISCL HBsAg kits and the correlation comparing with Roche kits.Methods A total of 580 serum samples were collected,HBsAg levels were detected by HISCL HBsAg kits and Elecsys HBsAg Ⅱ Quant kits,respectively.Data were analyzed,and the specificity,sensitivity and anti-jamming capability of HISCL kits were assessed by comparing with Roche kits.Results The intra-assay coefficients of variation (CV) of HISCL HBsAg kits was ＜ 5.0％,and Interassay CV was ≤5.5％.238 samples were repeated testing with HISCL HBsAg kits and the coincidence was 100％.Within them,the repeating detection results of 174 positive samples by HISCL HBsAg kits had agood correlation (r =0.999,P ＜0.01).122 cases of normal control were detected negative by both kits,and the coincidence was 100％.The correlation of HBsAg results from 287 HBsAg positive samples detected by the two kits was r =0.996,P ＜ 0.01.For the samples containing suspicious interferences,the results of the two kits were similar(pregnant women and infants r =0.992,P =0.000;antiviral drugs r =0.994,P =0.000;various virus or syphilis r =0.995,P =0.000;HBV gene mutation r =0.998,P =0.000;antinuclear antibodies and rheumatoid factor r =0.997,P =0.000).Conclusions HISCL HBsAg kits have a good repeatability and a excellent correlation with Elecsys HBsAg Ⅱ Quant kits.

Objective:To demonstrate the clinical effect of latissimus dorsi musculocutaneous flap with primary closure in V-Y suture in the repair of major lesions in the anterior chest wall that was left after mastectomies with locally advanced breast cancer (LABC) surgery.Methods:From September 2018 to February 2021, the technique was employed on 14 female cancer patients who had LABC surgery in the Department of Breast Surgery of the First Affiliated Hospital of Kunming Medical University. The patients received radical mastectomies with major resection of cutaneous tegument. The defect areas in chest wall were 15.0 cm×15.0 cm-22.0 cm×35.0 cm. The sizes of flap were 12.0 cm×28.0 cm-18.0 cm×35.0 cm. The sizes of musculocutaneous flap were 12.0 cm×28.0 cm×2.0 cm~18.0 cm×35.0 cm×3.5 cm. All patients were entered the postoperative follow-up through out-patient clinic and telephone interviews.Results:The flap provided an efficient coverage in closing the defects among all 14 patients. Three patients presented small areas (1.0-3.0 cm) of superficial necrosis in Y-cross area of the flap. None of the patient had back swelling. The average operation time was 6.3 hours. Postoperative follow-up varied from 4 to 41 months(18 months in average). The colour, texture, elasticity of the flaps were acceptable, with good shapes. Function of upper limbs was normal in 13 cases without lymphedema, except 1 who had lymphedema of affected limb at 3 years after surgery. Eleven cases had radiotherapy after surgery with good tolerance. None of the cases had local recurrence of breast cancer. Five cases had metastasis, 3 cases died of metastasis.Conclusion:The latissimus dorsi musculocutaneous flap with primary closure in V-Y suture is easy to perform and an reliable and efficient technique in repairing large defects in the anterior chest wall left after a LABC surgery.

Objective: To investigate the genetic characteristics of Lamivudine-resistant mutation patterns and HBV S gene mutants in patients with chronic hepatitis disease of different disease progression. Methods: Blood samples of LAM-resistant patients with chronic hepatitis disease were collected. HBV RT gene nucleotide sequences were obtained, and then differences in drug-resistant mutation patterns, drug susceptibility and HBV S gene mutants characteristics between the two groups were analyzed. Results: Forty-seven chronic hepatitis B (CHB) patients and 16 HBV-related liver cirrhosis (LC)/HBV-related hepatocellular carcinoma (HCC) patients were included in this study. M204I single point mutation and L180M+ M204I/V were the most common pattern during patients with chronic hepatitis disease (35/63, 55.56%). The numbers of resistant to three nucleos(t)ide analogues in LC/HCC group was higher than CHB group’s (62.50% vs 34.04%, P=0.046). In HBV S gene, more immune associated HBsAg-escape mutations were detected in LC/HCC group than that in CHB group (62.50% vs 31.91%, P=0.031). I126T/V and G145A (for LCC/HCC group, 60%), I126S/T and S117T (for CHB group, 46.67%) were showed as the most common form for HBsAg escape mutations in the two groups. The two groups both detected RT mutations concomitantly with stop codon mutations in S gene (rtA181T/sW172* and rtM204I/sW196*). Conclusions Different characteristics in Lamivudine-resistant mutations and associated HBV S gene mutants were found in patients with chronic hepatitis disease of different disease progression, and LC/HCC patients exhibit more multi-drug resistant variants and immune associated HBsAg-escape mutants than CHB patients.