Objective To investigate the relation between the human leucocytes antigen-DQA1(HLA-DQA1) alleles poly-morphism and the Uygur Han nationality hepatitis B patients in Xinjiang and the genetic susceptibility healthy controls ,which is provide some important clues to seek the susceptible genes and disease-resistant genes of HBV infection for Uygur and Han nation-ality hepatitis B patients .Methods HLA-DQAl alleles of 182 cases of the Hepatitis B patients and 163 people were compared with HBV DNA and ALT level .HLA-DQA1 * 0102 ,-DQA1 * 0104 ,-DQA1 * 0201 ,-DQA1 * 0301 ,-DQA1 * 0302 ,-DQA1 * 0501 genes frequency are detected with PCR-SSP .Results Compared the Uygur hepatitis B patients ALT abnormal and HBV DNA high copy quantity group with healthy controls group in allele′s frequency analysis found that HLA-DQA1 * 0301 ,-DQA1 * 0501 genes had statistic significance(P< 0 .05) .Compared the Han nationality hepatitis B patients ALT abnormal and HBV DNA high copy quanti-ty group with healthy controls group in allele′s frequency analysis ,It was found that HLA-DQA1 * 0102 ,-DQA1 * 0201 ,-DQA1 *0301 genes difference had statistic significance(P < 0 .05) .HLA-DQA1 * 0102 had the statistic significance between high and low copy quantity groups(P< 0 .05) .Compared the Han nationality hepatitis B patients ALT normal and low copy quantity group with healthy controls group in allele′s frequency analysis ,It was found that HLA-DQA1 * 0201 genes had statistic significance(P <0 .05) .HLA-DQA1 * 0102 ,-DQA1 * 0301 had statistic significance between the Han and Uygur nationality for HBV patients(P<0 .05) ;HLA-DQA1 * 0201 ,-DQA1 * 0501 had statistic significance between the Han and Uygur nationality for healthy people(P<0 .05) .Conclusion HLA-DQA1 * 0501 is the protected gene of Uygur hepatitis B patients ;-DQA1 * 0301 is the susceptibility gene .The Han nationality hepatitis B patients group HLA-DQA1 * 0102 ,-DQA1 * 0301 ,-DQA1 * 0302 is the susceptibility genes and -DQA1 * 0201 is the Antagonism gene .

In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

Objective To determine the value of ischemia modified albumin,heart-type fatty acid-binding protein,B-type natri-uretic peptide and homocysteine in the risk stratification of patients with unstable angina pectoris;thus to provide an assessment for the condition of patients in clinic.Methods 135 patients with unstable angina were included in the disease group and subjected to risk stratification according to GRACE risk score software,70 cases of low-risk group,60 cases in the middle-risk group and 5 cases in the high-risk group.Another 145 healthy people were in the control group.The levels of ischemia modified albumin,heart-type fatty acid-binding protein,B-type natriuretic peptide and homocysteine were detected and compared.Results Between the control group and the disease group,significant difference of heart-type fatty acid-binding protein,B-type natriuretic peptide and homocys-teine was found (P <0.05),but the difference of ischemia modified albumin was not statistically significant(P >0.05).In the dis-ease group,the levels of ischemia modified albumin,heart-type fatty acid-binding protein and homocysteine in each risk stratification showed no significant difference(P >0.05).The level of B-type natriuretic peptide in high-risk group was higher than that in the low-risk group and in the middle-risk group and the difference was statistically significant (P <0.05),while there was no statisti-cally significant difference between the low-risk group and the middle-risk group(P >0.05 ).Conclusion The detection of heart-type fatty acid-binding protein,B-type natriuretic peptide and homocysteine possesses certain meaning in diagnosing unstable angi-na,and the level of B-type natriuretic peptide indicates the risk degree of the disease.

The activity of the NK cells in patients with preeclampsia was studied to investigate the pathogenesis of preeclampsia. By using MTT and 51Cr releasing technique, the proliferation and killing ability of the NK cells in maternal and umbilical blood from preeclampsia patients (n = 18) and normal third trimester pregnant women (n = 18) were detected. The NK-92 cell line was as the positive control. The results showed that the NK cell counts of umbilical blood in preeclampsia patients and normal third trimester pregnant women were significantly greater than those of maternal blood (both P<0.05). Compared with that in normal third trimester pregnant women, the proliferative ability of the NK cells in preeclampsia patients was apparently increased (P<0.05). Compared with that in maternal blood, the proliferative ability of the NK cells in umbilical blood from both preeclampsia patients and normal third trimester pregnant women was dramatically increased. The killing ability of the NK cells in preeclampsia patients was significantly higher than that in normal third trimester pregnant women (P <0.05). It was suggested that both number and function of the NK cells in preeclampsia women were increased, and that in umbilical blood was greater than that in maternal blood, speculating that the function of the NK cells may affect the maintenance of the maternal and fetal immune tolerance during pregnancy.
Cytotoxicity, Immunologic/*immunology ; Fetal Blood/cytology ; Immune Tolerance ; Killer Cells, Natural/*immunology ; Killer Cells, Natural/pathology ; Pre-Eclampsia/blood ; Pre-Eclampsia/*immunology ; Pregnancy Trimester, Third

In order to investigate the effects of LPS on the TACE gene transcription and expression and its regulating effect on the TM-TNF secretion, in vitro studies were carried out on HL-60 cells stimulated by LPS. TACE, TNF-alpha mRNA levels were detected by Dot-Elisa and the distribution of membrane molecules determined by flow cytometry assay and indirect immunofluorescence. The results showed that: (1) TACE was detected in or on HL-60 cells and it is predominantly localized on cell surface and to a perinuclear compartment. (2) LPS induced a time dependent increasement of TNF-alpha mRNA and enhanced TNF conversion with decreasing distribution of TNF in cell surface and increasing secretion of TNF protein. Such conversion could be inhibited by TACE ODN. (3) LPS also induced time-dependently increased expression of TACE gene and activation of its function. On the other hand, TACE protein in cell lysate and on cell surface was decreased. It was suggested that TACE molecular structure might change following its mediating membrane-anchored molecular secretion.
ADAM Proteins ; ADAM17 Protein ; Gene Expression ; HL-60 Cells ; Humans ; Lipopolysaccharides ; pharmacology ; Metalloendopeptidases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transcription, Genetic ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

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.

Background::Pathological scars are a disorder that can lead to various cosmetic, psychological, and functional problems, and no effective assessment methods are currently available. Assessment and treatment of pathological scars are based on cutaneous manifestations. A two-photon microscope (TPM) with the potential for real-time non-invasive assessment may help determine the under-surface pathophysiological conditions in vivo. This study used a portable handheld TPM to image epidermal cells and dermal collagen structures in pathological scars and normal skin in vivo to evaluate the effectiveness of treatment in scar patients. Methods::Fifteen patients with pathological scars and three healthy controls were recruited. Imaging was performed using a portable handheld TPM. Five indexes were extracted from two dimensional (2D) and three dimensional (3D) perspectives, including collagen depth, dermo-epidermal junction (DEJ) contour ratio, thickness, orientation, and occupation (proportion of collagen fibers in the field of view) of collagen. Two depth-dependent indexes were computed through the 3D second harmonic generation image and three morphology-related indexes from the 2D images. We assessed index differences between scar and normal skin and changes before and after treatment.Results::Pathological scars and normal skin differed markedly regarding the epidermal morphological structure and the spectral characteristics of collagen fibers. Five indexes were employed to distinguish between normal skin and scar tissue. Statistically significant differences were found in average depth ( t = 9.917, P <0.001), thickness ( t = 4.037, P <0.001), occupation ( t= 2.169, P <0.050), orientation of collagen ( t = 3.669, P <0.001), and the DEJ contour ratio ( t = 5.105, P <0.001). Conclusions::Use of portable handheld TPM can distinguish collagen from skin tissues; thus, it is more suitable for scar imaging than reflectance confocal microscopy. Thus, a TPM may be an auxiliary tool for scar treatment selection and assessing treatment efficacy.

The activity of the NK cells in patients with preeclampsia was studied to investigate the pathogenesis of preeclampsia. By using MTT and 51Cr releasing technique, the proliferation and killing ability of the NK cells in maternal and umbilical blood from preeclampsia patients (n = 18) and normal third trimester pregnant women (n = 18) were detected. The NK-92 cell line was as the positive control. The results showed that the NK cell counts of umbilical blood in preeclampsia patients and normal third trimester pregnant women were significantly greater than those of maternal blood (both P<0.05). Compared with that in normal third trimester pregnant women, the proliferative ability of the NK cells in preeclampsia patients was apparently increased (P<0.05). Compared with that in maternal blood, the proliferative ability of the NK cells in umbilical blood from both preeclampsia patients and normal third trimester pregnant women was dramatically increased. The killing ability of the NK cells in preeclampsia patients was significantly higher than that in normal third trimester pregnant women (P <0.05). It was suggested that both number and function of the NK cells in preeclampsia women were increased, and that in umbilical blood was greater than that in maternal blood, speculating that the function of the NK cells may affect the maintenance of the maternal and fetal immune tolerance during pregnancy.
Adult ; Cytotoxicity, Immunologic ; immunology ; Female ; Fetal Blood ; cytology ; Humans ; Immune Tolerance ; Killer Cells, Natural ; immunology ; pathology ; Pre-Eclampsia ; blood ; immunology ; Pregnancy ; Pregnancy Trimester, Third

Objective:To explore the clinical features and survival of newly diagnosed follicular lymphoma (FL) patients with diffuse large B-cell lymphoma (DLBCL) component.Methods:1845 newly diagnosed FL patients aged ≥ 18 years with grades 1-3a in 11 medical centers in China from 2000 to 2020 were included, and patients with DLBCL component were screened. The clinical data and survival data of the patients were retrospectively analyzed, and the prognostic factors were screened by univariate and multivariate analysis.Results:146 patients (7.9% ) with newly diagnosed FL had DLBCL component. The median age was 56 (25-83) years, 79 males (54.1% ) . The pathology of 127 patients showed the proportion of DLBCL component. Patients were divided into two groups according to whether the proportion of DLBCL component was ≥ 50% . The study found that patients with DLBCL component ≥ 50% had higher grade 3 ratio (94.3% vs 91.9% , P=0.010) , Ki-67 index ≥ 70% ratio (58.5% vs 32.9% , P=0.013) and PET-CT SUVmax ≥ 13 ratio (72.4% vs 46.3% , P=0.030) than patients with DLBCL component<50% . All patients received CHOP or CHOP like ± rituximab chemotherapy. The overall response rate (ORR) was 88.2% , and the complete response (CR) rate was 76.4% . In the groups with different proportions of DLBCL component, there was no significant difference in the remission rate after induction treatment and the incidence of disease progression within 2 years after initiation of treatment (POD24) ( P<0.05) . The overall estimated 5-year progression free survival (PFS) rate was 58.9% , and the 5-year overall survival (OS) rate was 90.4% . The 5-year OS rate of POD24 patients was lower than that of non POD24 patients (70.3% vs 98.5% , P<0.001) . Compared with non maintenance treatment of rituximab, maintenance treatment of rituximab could not benefit the 5-year PFS rate (57.7% vs 58.8% , P=0.543) , and the 5-year OS rate had a benefit trend, but the difference was not statistically significant (100% vs 87.8% , P=0.082) . Multivariate analysis showed that failure to reach CR after induction treatment was an independent risk factor for PFS ( P=0.006) , while LDH higher than normal was an independent risk factor for OS ( P=0.031) . Conclusion:FL patients with DLBCL component ≥50% have more invasive clinical and pathological features. CHOP/CHOP like ± rituximab regimen can improve the clinical efficacy of patients. Rituximab maintenance therapy can not benefit the PFS and OS of patients. Failure to reach CR after induction therapy was the independent unfavorable factor for PFS.