Objective To evaluate the expression of Toll-like receptor(TLR)on the monocyte- derived dendritic cells(DC)from chronic hepatitis B(CHB)patients and to analyze the expression pro- file and significance of the TLR such as TLR3,TLR4,TLR?,TLR8 and TLRg,which are associat- ed with immune response to viral infection.Methods Peripheral blood mononuclear cell(PBMC) centrifugated by the hydroxyethyl starch(HES)centrifugation were cultured and induced into DC by granulocyte-maerophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4),and their mor- phology and phenotype were detected by the inverted microscope and flow cytometry respectively. Monocyte-derived DC were obtained from 10 chronically hepatitis B virus(HBV)-infected patients and 15 healthy volunteers.TLR3,TLR4,TLR7,TLRS,TLR9 expression on immature and mature DC were analyzed by FACS Calibur.DC was pulsed with HBcAg on day 3 and 5,then DC maturation and ability to process HBcAg and to stimulate autogeneic T cells were evaluated.Results Monocyte- derived DC developed different TLR expression patterns as they went through different maturation stages.TLR7,TLR8 expressions on immature DC and TLR3,TLR7 expressions on mature DC were lower in CHB than in control(for TLR7,TLR8 expression on immature DC:75.9%,1.0%vs 98.4%,15.4%,P

Abstract: Objective　To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods　According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results　The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion　The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.

Objective To study the molecular mechanisms of ampicillin- resistant haemophilius influenzae (Hi)in Nanjing. Methods One hundred and fifty- eight strains of Hi isolated from children were collected to detect bata-lactamase. TEM and ROB bata- lacta-mase genes were detected by polymerase chain reaction (PCR) ,and cloned into T vector for sequencing. Results The rate of ampicillin resistance was 41. 77% in Hi isolated from children in Nanjing,40.51 % was found to be bata-lactamase production. Eighty-nine strain were TEM positives, 1 strain was ROB positive,63 strains bata - lactamase positive ampicillin- resistant Hi were identified. The resistance mechanism of ampicillin resistant Hi was production of bata - lactamase , mainly TEM - type enzyme. Two bata - lactamase negative ampicillin - resistant Hi were identified , predicts the other mechanisms of ampicillin - resistant Hi was occuered yet . One strain of non -TEM - type,and non - ROB - type bata - lactamase - producing Hi was identified. Conclusions Ampicillin - resisitant in Hi isolated from children in this region is challenging. TEM bata - lactamase is the principal mechanism of ampicillin - resistant of Hi.

Animals ; Carcinoma, Hepatocellular ; blood supply ; virology ; Cell Transformation, Neoplastic ; Endothelial Growth Factors ; biosynthesis ; genetics ; Gene Expression Regulation, Viral ; Hepatitis B virus ; pathogenicity ; physiology ; Humans ; Liver Neoplasms ; blood supply ; virology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Trans-Activators ; physiology

We report here on a rare case of an ectopic pancreatic tissue in the anterior mediastinum. A 32-year-old woman without any symptoms was transferred to our hospital because of an abnormal large mediastinal shadow on her chest radiograph during a checkup. The computed tomography (CT) scan revealed a giant cystic-solid mass that measured 16 x 13 x 8 cm and it was located in the center of the anterior mediastinum and it symmetrically grew to two sides. On enhanced CT scans, the solid component of the mass showed marked enhancement. We performed total surgical resection of the mass and complete pancreatic tissues were verified on the pathological examination.
Adult ; Choristoma/*radiography/surgery ; Diagnosis, Differential ; Female ; Humans ; Mediastinal Diseases/*radiography/surgery ; *Pancreas ; Tomography, X-Ray Computed

OBJECTIVETo study the expression and clinical significance of ribosomal S6 kinase-4 (RSK-4) in breast cancer and explore the role of RSK-4 in the genesis and development of breast cancer.

METHODThe expression levels of RSK-4 mRNA and protein were detected in 56 cases of breast cancer and the normal breast tissues, as well as in 20 cases of breast benign lesions, by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry.

RESULTSThe expression rates of RSK-4 mRNA in breast cancer, the normal breast tissues and breast benign lesions were 48.2%, 76.8% and 75.0%, respectively. The expression level of RSK-4 mRNA in breast cancer was significantly lower than those in normal breast tissues and breast benign lesions tissues (P < 0.05). The expression level of RSK-4 significantly correlated with tumor size and clinical stage (P < 0.05).The expression rate of RSK-4 protein was 39.3% in breast cancer tissues, which was significantly lower than that of normal breast tissues (71.4%) and breast benign lesions (75.0%, P < 0.01). The expression level of RSK-4 protein was lower in breast cancer with large tumor, high clinical stage and lymph node metastasis. In 56 cases of breast cancer samples, the consistency rate of RSK-4 mRNA and protein was 73.2%. A significant correlation was found between RSK-4 mRNA and protein (χ² = 10.254, P < 0.05).

CONCLUSIONThe down-regulation of RSK-4 expression in breast caner suggests that it is a breast cancer suppressor gene, and the lack or down-regulation of RSK-4 expression is involved in the genesis and progression of breast cancer.

Adult ; Aged ; Breast ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Down-Regulation ; Female ; Fibroadenoma ; metabolism ; pathology ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Ribosomal Protein S6 Kinases, 90-kDa ; genetics ; metabolism ; Tumor Burden ; Young Adult

OBJECTIVETo develop a specific quality of life (QOL) instrument for Chinese patients living with HIV/AIDS and to assess its reliability and validity.

METHODSA primary questionnaire was developed with reference to existing instrument and information through discussion with experts, front-line professionals and patients living with HIV/AIDS. This questionnaire was applied in 443 patients living with HIV/AIDS. Some items were removed through the results of responsive rate, factor analysis, correlation matrix,internal consistency and so on. Reliability and validity were assessed after items reduction.

RESULTSThe developed instrument contained 44 items, including 10 scales and other 3 items used for the evaluation of curative effect and side effect of anti-virus drug. No substantial ceiling/floor effects existed. All scales' internal consistency coefficients were > or = 0.70 and the whole instrument was 0.90. The two week retest reliability coefficient of the whole instrument was 0.80, and there were 6 scales below 0.70 of this coefficient. Multitrait/multiitem assessment indicated scaling success rates were all 100% for all items. Spearman correlation coefficient of total score of this instrument with SF-36 was 0.69.

CONCLUSIONIt was the first time that the developed QOL-CPLWHA instrument introducing the scale of hostility regarding mentality trend as well as expressed its acceptable reliability and validity so could be used for the evaluation of Chinese patients living with HIV/AIDS.

Acquired Immunodeficiency Syndrome ; psychology ; Adolescent ; Adult ; Aged ; China ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo assess the quality of life (QOL) and hostile mentality trend (HMT) of 299 patients living with HIV/AIDS (Human immunodeficiency virus/Acquired immune deficiency syndrome) in three provinces in China, and to understand the major concerns of the these patients.

METHODSThe SF-36 (short form -36) was used for assessing the QOL among 299 HIV-infected patients in Sichuan, Hubei and Guizhou provinces. Reliability and validity of SF-36 were evaluated. Consulting with experts and professionals, seven additional items were developed to evaluate the HMT. Mean scores of the 8 scales were compared between the patients and general rural residents in Sichuan province.

RESULTSFor SF-36, internal consistent coefficients (Cronbach's alpha) of the 8 scales were between 0.75 to 0.90, test-retest reliability coefficient ranged from 0.54 to 0.80. The item-subscale correlation coefficients ranged from 0.46 to 0.97. Mean scores of the 8 scales of the patients ranged from 28.50 to 77.87, and 70.27 to 91.87 for the general rural residents. The variations of the scales were tested by means of Mann-Whitney test with u value ranged from -17.43 to -23.87. The QOL of the patients living with HIV/AIDS were significantly inferior to those of general population (all P < 0.01). The mean scores of the seven items to evaluate HMT ranged from 46.21 to 82.89. The major concerns of the patients living with HIV/AIDS included financial insecurity and family responsibilities, followed by death threat and no cure of HIV/AIDS.

CONCLUSIONThe SF-36 is a reliable instrument for assessing QOL of patients living with HIV/AIDS. The QOL of the patients living with HIV/AIDS in China is poor.

Acquired Immunodeficiency Syndrome ; psychology ; Adolescent ; Adult ; Aged ; Educational Status ; Female ; HIV Infections ; psychology ; Hostility ; Humans ; Male ; Marital Status ; Middle Aged ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; standards ; Young Adult

OBJECTIVEThe aim of this study was to investigate the changes in mTOR activity and survivin expression in liver cancer cell line HepG2 cells treated with tamoxifen.

METHODSSurvivin transcription level and p70S6K was demonstrated by PCR, dual-luciferase reporter assay and Western blot analysis, respectively, and the apoptosis in the HepG2 cells was detected by flow cytometry.

RESULTSTamoxifen leads to apoptosis of the cells and reduction in survivin expression, as well as a dramatic reduction in the activated form of p70S6K. Treating HepG2 cells with rapamycin, a specific mTOR inhibitor, significantly reduced the survivin protein level but not affected the survivin transcription, indicating that tamoxifen and rapamycin were synergistic in regards to down-regulation of survivin expression in hepatocellular carcinoma cells.

CONCLUSIONSOur results suggest that tamoxifen down-regulates survivin expression in HepG2 cells and it is mediated by transcriptional and post-transcriptional level via PI3K/Akt/mTOR pathway to induce apoptosis.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents, Hormonal ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; Drug Synergism ; Hep G2 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; Sirolimus ; pharmacology ; TOR Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; Tamoxifen ; pharmacology

OBJECTIVETo investigate the effects of HBx gene on proliferation activity of hepatoma cells in vitro and in vivo.

METHODSThe plasmid pHA-HBx carrying HBx gene was transfected into HepG(2) cells, and the positive clones were screened and identified with G418 and RT-PCR, respectively. The growth curve and population doubling time were calculated, and the cell cycle was analyzed by flow cytometry (FCM). The proliferation activity of transformed cells was measured with (3)H-TdR incorporation rate and nude mice model in vitro and in vivo.

RESULTSThe result of RT-PCR indicated that HBx gene was integrated into the genome DNA of HepG(2) cells and transcripted. The growth curve and population doubling time showed a high proliferation activity of transformed cells. The amount of cells at stage S and G(2)/M were significantly higher, and cells at stage G(0)/G(1) were lower than those in control group. The tumors developed from transfected cells grew much quicker than those developed from HepG(2) cells in nude mice model.

CONCLUSIONHBx gene can facilitate the proliferation of hepatoma cells both in vitro and in vivo.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Cycle ; genetics ; Cell Division ; genetics ; Cell Line, Tumor ; Female ; Flow Cytometry ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection ; Transplantation, Heterologous