Expanded circulating peripheral helper T cells in primary biliary cholangitis: Tph cells in PBC.

BACKGROUND: Peripheral helper T (TPH) cells, a recently defined subset of Th cells, promote B cell differentiation and antibody production in inflamed tissues. This study investigated whether circulating TPH cells are associated with primary biliary cholangitis (PBC), a typical organ-specific autoimmune disease. METHODS: Twenty PBC patients and 20 age- and sex-matched healthy controls (HCs) were recruited. The circulating TPH cell subsets were analyzed by flow cytometry, and the associations of TPH cells with disease activity and plasma cells were determined. Functional analysis was performed using a TPH and B cell coculture experiment. RESULTS: The frequencies of circulating TPH cells, ICOS+ TPH cells, and CD28+ TPH cells were increased in patients with PBC. Furthermore, the ICOS+ TPH cell level was higher in PBC patients with or without cirrhosis than in HCs, and the level decreased after treatment. Moreover, ICOS+ TPH cell levels correlated positively with specific clinical parameters (including anti-mitochondrial antibodies against M2 antigen (AMA-M2), IgM) and plasma cell levels, suggesting that the TPH cell activation status is associated with the severity of PBC. Coculture results revealed an enhanced ability of TPH cells from PBC patients to induce B cell differentiation. CONCLUSIONS: Elevated numbers of TPH cells may be involved in the pathogenesis of PBC, and the activation status of TPH cells is related to the severity of PBC. Additionally, TPH cells can be used as a useful biomarker for evaluating the progression of PBC and may serve as a therapeutic target for PBC patients in the future.

Clinical Data on Hospital Environmental Hygiene Monitoring and Medical Staffs Protection during the Coronavirus Disease 2019 Outbreak

BackgroundThe outbreak of coronavirus disease 2019 (COVID-19) has placed unprecedented challenges on hospital environmental hygiene and medical staff protection. It is crucial to assess hospital environmental hygiene to understand the most important environmental issues for controlling the spread of COVID-19 in hospitals. ObjectiveTo detect the presence of COVID-19 in the samples from the area at risk of contamination in the First Hospital of Jilin University. MethodsViruses in the air were collected by natural sedimentation and air particle sampler methods. Predetermined environmental surfaces were sampled using swabs at seven oclock in the morning before disinfection. The real-time reverse-transcription PCR method was used to detect the existence of COVID-19 pathogens. ResultsViruses could be detected on the surfaces of the nurse station in the isolation area with suspected patients and in the air of the isolation ward with an intensive care patient. ConclusionComprehensive monitoring of hospital environmental hygiene during pandemic outbreaks is conducive to the refinement of hospital infection control. It is of great significance to ensure the safety of medical treatment and the quality of hospital infection control through the monitoring of environmental hygiene.

Non-operative Treatment for Acute Intra-synovial Sheath ACL Rupture / 中国运动医学杂志

Objective To evaluate the effectiveness of non-operative treatment for the acute intra-synovial sheath anterior cruciate ligament (ACL) rupture.Methods Twenty-eight patients diagnosed as the acute intra-synovial sheath ACL rupture at outpatient clinic between May 2014 and July 2016 were included.All patients were immobilized with knee braces for 6 weeks,followed by range of motion (ROM) training and partial to full weight-bearing of knees.All patients returned 3 months later for MRI scanning and those with the side-to-side difference of the anterior-posterior laxity less than 5 mm continued with non-operative treatment,followed up for MRI examination and clinical assessments 6 and 12 months later.Results Four patients dropped out because they didn't meet the stability criteria at 3 months after the treatment,3 of whom received surgical reconstruction and 1 with muscle strengthening training.Another patient received surgical reconstruction at 5 months due to re-injury.The remaining 23 patients achieved satisfactory results at 12 months after the treatment,with the average side-to-side difference of the anterior-posterior laxity of 2.1mm (0-4 mm),MRI good-to-excellent rate of 85.2％ (8 of Grade 1 and 15 of Grade 2),subjective IKDC (International Knee Documentation Committee) score of 92.71 (89.7-98.9),Lysholm score of 91.6 (86-95),and modified Larson score 96.4 (92-99).Conclusions Patients with the acute intra-synovial sheath anterior cruciate ligament (ACL) rupture showed satisfactory functional scores and objective stability and healing on MRI after the non-operative treatment.

Research on the Relationship between the Anterior Ridge and the Tibial Attachment of the Anterior Cruciate Ligament on Magnetic Resonance Imaging / 中国运动医学杂志

Objective To measure and determine the position of the tibial attachment of the anterior cruciate ligament (ACL)in relation to its anterior ridge on the magnetic resonance imaging (MRI)of normal knee joints,and to explore the clinical role of the anterior ridge in guiding tunnel positioning during ACL reconstruction as well as in facilitating postoperative radiographic evaluation.Methods The knee MRI of one hundred young adult patients with an intact ACL and normal knee joint (mean age:25.1 years,range:18-40 years)was retrospectively reviewed.All MR images were obtained at full extension of the knee on the same MRI machine.Using digital image software on MRI,the measurements in the sagittal view were taken,including the depth of the tibia,the distance from the anterior edge of the tibial plateau to the most anterior and posterior portions of the ACL insertion on the tibia and the anterior ridge according to Staubli and Rauschning's method.Results The center of the tibial insertion of the ACL is located between 28.43％ and 50.94％ of the total anterior-posterior depth of the tibia,which was less than 43.3％ in 58 patients.The average distance from the anterior edge of the tibial plateau to the anterior ridge was 13.61 ± 2.17 mm (ranging from 8.03 to 18.65 mm),26.80％ ±3.89％ (ranging from 17.74％ to 33.94％)across the tibial plateau.There were significant positive correlations between the distance from the anterior edge of the tibial plateau to the most anterior portion of the ACL insertion and that to the anterior ridge.The distance from the most anterior portion of the ACL insertion to the anterior ridge was averaged 0.56 ± 0.68 mm (ranging from-0.28 to 2.71 mm).During the ACL reconstruction,with the anterior edge of the tibial tunnel determined at posterior 0.5 mm to the anterior ridge,the graft size as 8 mm,and the tibial guider angle set as 55 degree,96of the patients (96％)would have the center of the tibial tunnel located before the center of their native ACL attachment.Conclusions On sagittal MR images,the location of the anterior ridge and the most anterior portion of the ACL insertion correlated well,with the average distance between them of 0.56 mm.The study indicates that during ACL reconstruction,tibial tunnel drilling with the anterior edge of the ACL graft positioned at the anterior ridge can achieve a more anterior position than the traditional methods to orientate according to the center of the bone tunnel.

The progress and prospect of application of genetic testing technology-based gene detection technology in the diagnosis and treatment of hereditary cancer / 中华预防医学杂志

Hereditary cancer is caused by specific pathogenic gene mutations. Early detection and early intervention are the most effective ways to prevent and control hereditary cancer. High-throughput sequencing based genetic testing technology (NGS) breaks through the restrictions of pedigree analysis, provide a convenient and efficient method to detect and diagnose hereditary cancer. Here, we introduce the mechanism of hereditary cancer, summarize, discuss and prospect the application of NGS and other genetic tests in the diagnosis of hereditary retinoblastoma, hereditary breast and ovarian cancer syndrome, hereditary colorectal cancer and other complex and rare hereditary tumors.

Helicobacter pylori Infection Increases Frequency of PDCA-1(+) (CD317(+)) B-cell Subsets.

BACKGROUND AND AIMS: As a newly discovered B-cell subset, PDCA-1(+) B cells have been shown to participate in the immune clearance of invading pathogens. The prominence of PDCA-1(+) B cell immunity in the pathogenesis of Helicobacter pylori infection prompted us to explore the potential role of this subset in gastric H. pylori infection. METHODS: H. pylori infection was determined by (14)C-urea breath test and Western blot. The frequency of the different sub-compartments of PDCA-1(+) B cells and their relation to serum cytokines was determined in 33 H. pylori-infected and 14 uninfected patients and in 12 healthy controls (HC). RESULTS: In comparison to uninfected individuals, there was a significantly increased frequency of PDCA-1(+) B cells, PDCA-1(+)IgM(+) B cells, CD93(+)PDCA-1(+) B cells, CD93(+)PDCA-1(+)IgM(+) B cells, CD137(+)PDCA-1(+) B cells and CD137(+)PDCA-1(+)IgM(+) B cells were detected in patients with H. pylori infection, corresponding to increased levels of serum IFN-α and IgM in this group. Compared with H. pylori-positive (HP(+)) chronic non-atrophic gastritis patients, a larger proportion of PDCA-1(+) B cells, CD93(+)PDCA-1(+) B cells and CD137(+)PDCA-1(+) B cells were observed in HP(+) patients suffering from atrophic gastritis or HP(+) peptic ulcers. CONCLUSIONS: The frequency of the PDCA-1(+) B cell compartment is increased during H. pylori infection. Our data support the potential role of this B-cell subset in the pathogenesis of H. pylori-dependent gastritis.

Effect of anterior cruciate ligament rupture on hamstring∶quadriceps ratio during isokinetic knee extension and flexion at 30 degrees of flexion / 北京大学学报(医学版)

Objective:To evaluate the change in hamstring ( H ):quadriceps ( Q ) ratio following anterior cruciate ligament ( ACL) rupture during isokinetic knee extension and flexion at 30 degrees of flexion which is important for knee dynamic function .Methods:A study was performed in 25 male com-plete unilateral ACL ruptures .Isokinetic concentric and eccentric quadriceps and hamstring muscle tests in both the deficient knees and intact knees were performed at 60°/s, respectively.At 30 degrees of flexion, the average torque of quadriceps and hamstring , Qe∶Qc ratios ( ratios of eccentric quadriceps to concentric quadriceps muscle torque ) , He∶Hc ratios ( eccentric hamstring to concentric hamstring ) , Hc∶Qc ratios ( concentric hamstring to concentric quadriceps ) , He∶Qc ratios ( eccentric hamstring to con-centric quadriceps ) , and Hc∶Qe ratios ( concentric hamstring to eccentric quadriceps ) were calculated . Wilcoxon matched-pairs signed-ranks test was used .Results:At 30 degrees of knee flexion , a significant reduction ( P<0.05) in the average torque of quadriceps was observed at concentric and eccentric 60°/s produced by the deficient-side compared with the intact side .In addition, Hc∶Qc, He∶Qc, and Qe∶Qc significantly increased on the ACL-deficient side .Conclusion:The change in H ∶Q ratio in the mode of isokinetic 60 °/s at 30 degrees of knee flexion might therefore be a new tool to objectively document muscle function in ACL-deficient knee .

Construction of recombinant adenovirus vector pAV.Ex1 d-Hiwi using Gateway technology / 吉林大学学报(医学版)

Objective To obtain the Hiwi gene encoding full-length and construct human Hiwi adenoviral vectors carrying green luorescence protein (GFP), and to establish foundation for a further study on Hiwi function and mechanism of inducing leukemia stem cell differentiation and apoptosis.Methods All coding areas of human Hiwi gene full length were amplified using method of overlapping extension PCR technology, and the full length coding aeras were inserted into the vector of Flag-IRES-hrGFP carrying GFP with Gateway technology to construct pDown-Hiwi-3 × flag-IRES-hrGFP. The cloning vector pDown-Hiwi-3 × flag-IRES-hrGFP and expression vector pAV. Des1d were used for homologous recombination reaction to obtain recombinant adenovirus vector pAV.Ex1d-Hiwi-3× flag-IRES-hrGFP.The positive clones were selected by PCR to extract the recombinant adenovirus plasmid and to pack into recombinant Ad-Hiwi-3 × flag-IRES-hrGFP adenovirus. Results The human recombinant Hiwi was successfully cloned and the recombinant adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was found to be successfully constructed via restriction enzyme digestion and sequencing methods. The adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was transfected into the HEK293A cells using lipofectamine mediated gene transfection method. Under fluorescence microscope, the transfected cells with green fluorescence could be observed.Conclusion The expression plasmid of adenovirus vector pAV.Ex1d-Hiwi-3 × flag-IRES-hrGFP is successfully constructed and it can express in HEK293A cells.

Relationship between serum aminotransferase levels and metabolic disorders in northern China.

BACKGROUND/AIMS: Increasing evidence suggests an association between elevated serum aminotransferase levels and metabolic disorders (metabolic syndrome, hyperlipidemia and diabetes mellitus). However, the significance of relatively low levels of aminotransferases in relation to metabolic disorders has not been fully investigated in the general population. We investigated the association between serum aminotransferase levels and metabolic disorders using data from a survey in Jilin Province, China. MATERIALS AND METHODS: In 2007, a prospective survey was conducted throughout Jilin, China, covering both urban and rural areas. A total of 3835 people, 18-79 years old, were undergoing real-time ultrasonography, blood tests, and interviews with a structured questionnaire. RESULTS: Serum aminotransferase levels within the normal range were associated with metabolic syndrome independent of age, occupation, cultural and educational level, income, body mass index, waist circumference, smoking, and alcohol intake. Compared with the lowest level (< 20 IU/L), the adjusted odds ratios for alanine aminotransferase levels of 20-29, 30-39, 40-49, and >50 IU/L were 1.92, 2.50, 2.97, and 3.52 in men, and 1.38, 1.54, 3.06, and 2.62 in women, respectively. Near-normal serum aminotransferase levels associated with hyperlipidemia, non-alcoholic fatty liver disease, and diabetes mellitus were also found in the study. CONCLUSIONS: Normal to near-normal serum aminotransferase levels are associated with metabolic disorders. Serum alanine aminotransferase levels of 21-25 IU/L for men and 17-22 IU/L for women are suggested as cut-off levels that detect metabolic disorders affecting the liver.

Anti-proliferative effect of dehydroepiandrosterone and its metabolites on human tumor lines / 吉林大学学报(医学版)

Objective To study the inhibitory effects of dehydroepiandrosterone (DHEA) and its metabolites-dehydroepiandrosterone sulfate (DHEAs) on the proliferation of HepG2 and HT-29 and their mechanism.Methods HepG2 and HT-29 were incubated by DHEA or DHEAs with different concentrations (1,10,50,100 and 200 ?mol?L-1) for 8,24,48,72 h and routine culture was used as control.The inhibitory rate was detected by using MTT chromometry and BrdU assay respectively.The activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR),glucose -6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were examined simultaneously.Results ①MTT chromometry:DHEA with different concentrations obviously inhibited the growth of HepG2 and HT-29 cells compared with control group(P0.05).②BrdU assay:the growth of cells were significantly inhibited by DHEA with concentrations of 50,100 and 200 ?mol?L-1,especially to HepG2 cells(P0.05).Conclusion DHEA has strong anti-proliferative effects on both HepG2 and HT-29 cell lines and inhibitory effects on the activities of G6PD or HMGR,however,DHEAs has no obvious effect.

Construction of eukaryotic expression vector of hepatitis C virus NS5A gene / 吉林大学学报(医学版)

Objective To construct an eukaryotic expression vector of the hepatitis C virus(HCV)NS5A gene.and obtain a stable transfected Huh-7 cell line which provides a basis for further investigation of the hepatitis virus C NS5A protein.Methods The HCV NS5A gene from the pcDNA3.1(+)/HCV NS345 plasmid with HCV NS5A gene was amplified by PCR,and cloned to pGEM-T vector,and transformed into E.coli JM109.The positive colonies were first confirmed by restriction enzyme digestion and sequencing and then were inserted to eukaryotic expression vector pCI-neo,and verified by enzyme digestion and sequencing.Results After TA colon of the HCV NS5A was amplified by PCR,the positive colonies were finally verified by sequencing,which were totally in line with the designed coding sequence of HCV NS5A gene.Conclusion Eukaryotic expression vector of HCV NS5A gene has been successfully constructed.

Influences of 5-azacytidine on mesenchymal stem cells proliferation and differentiation into myoblasts / 吉林大学学报(医学版)

Objective To investigate the influences of 5-azacytidine (5-Aza) with different concentrations on mesenchymal stem cells (MSCs) proliferation and differentiation into myoblasts. Methods Bone marrow-derived MSCs of 4 weeks Wistar rats were separated and purified, and then treated with 5-Aza with different concentrations. The growth ability of cell was assayed with methyl thiazolyl tetrazolium (MTT) method. The expression of skeletal muscle actin in MSCs was determined with reverse transcription polymerase chain reaction (RT-PCR) and electrophoresis after MSCs were induced. Results The cell proliferation was not affected by 0 and 1 ?mol ? L-1 5-Aza . There were expressions of skeletal muscle actin treated with 3 -12 ?mol ? L-1 5-Aza. Some cells were obviously enlarged at the 9th day after induction and myotubu-like cells were found at the 12th day when treated with 9 and 12 ?mol ? L-1 5-Aza. 20 - 30 ?mol ? L-1 5-Aza induced the toxic effect on proliferation of MSCs. With the increase of concentration, the proliferation ability of MSCs was weakened. Conclusion 5-Aza affects the expression of regulatory gene to the stem cells and regulate MSCs to orientationally differentiate into myotube-like cells.

Expression levels of CD4~+CD25~+ regulatory T cells in peripheral blood of patients with chronic hepatitis B and clinical significance / 吉林大学学报(医学版)

Objective To investigate the proportion of CD4+CD25highTreg and CD4+CD25+CD127low/-Treg in peripheral blood in patients with chronic hepatitis B(CHB) and determine the relationship between the proportion of CD4+CD25+Treg and clinical parameters.Methods Fresh isolated peripheral blood mononuclear cells(PBMCs) of 28 patients with CHB and 24 healthy donors were analyzed for the proportion of CD4+CD25+Treg using flow cytometry by surface staining for CD4-PC5,CD25-FITC,CD127-PE.HBsAg,HBsAb,HBeAg,HBeAb and HBcAb were evaluated.HBV DNA levels were measured using real-time RT-PCR.Results The proportions of CD4+CD25highTreg to CD4+ T cells(3.36%?2.59%) and CD4+CD25+CD127low/-Treg(4.05%?1.63%) to CD4+ T cells in patients with CHB were higher than those in health controls(1.60%?0.66%,1.75%?0.83%,P100U?L-1) had a higher fraction(4.26%?3.10%) of CD4+CD25highTreg in peripheral blood than those patients with low level ALT(ALT

Determination of CD4~+CD25~+ regulatory T cells from peripheral blood in patients with multiple organ dysfunction syndrome and clinical significance / 吉林大学学报(医学版)

Objective To explore the change of CD4+ CD25+ regulatory T cells(Treg) from peripheral blood in patiens with multiple organ dysfunction syndrome(MODS) before and after treatment and its significance.Methods The frequencies of CD4+ CD25+ CD127-Treg were detected in 10 patients with MODS respectively before treatment and 1 week after treatment and 10 healthy donors by flow cytometry labeled with specific fluorescent antibodies,such as anti-CD4(PE-CY5),anti-CD25(FITC) and anti-CD127(PE).Results The frequency of CD4+ CD25+ CD127-Treg in patients after treatment(2.11%?0.33%) was significantly lower than before treatment(7.44%?1.59%)(P

Expression of Human Interleukin-33 and studies on its biology function / 中国免疫学杂志

Objective:To clone human interleukin-33(hIL-33)and express it in E.coli efficiently.Methods:The primers were synthesized according to the hlL-33 cDNA sequence in GeneBank.The hIL-33 was amplified by RT-PCR from human fibroblast cell line(L929),the PCR product was inserted into pUC19 vector.The IL-33 cDNA confirmed by sequencing was inserted into expressing vector PQE30 and expressed in E.coli M15 strain.IL-33 protein expression was induced by IPTG and purified by Ni-NTA affinity chromatography.The recombinant IL-33 was identified by Immunoblot and its biological activity was analyzed.Results:DNA sequencing confirmed that the cloned cDNA was identical to the published sequence of hIL-33.The recombinant plasmid PQE/hIL-33 was transformed into M15.An expected 18KD protein of hIL-33 found mainly in the induced host strains about 25% of total bacteria lysis by SDS-PAGE and coomassie blue staining.The 18 KD protein could be recognized by anti-IL33 antibody in western blot.The recombinant protein was purified to more than 95% of total protein and induces the production of IL-4 and IL-5 in human peripheral blood mononuclear cells.Conclusion:We have successfully expressed hIL-33 protein in E.coli and the expressed product has IL-33 specific bioactivity.