Objective:To investigate influences of two different HLA-B antigens expressed on K562 cells on receptors expression of NK cells from peripheral blood lymphocytes.Methods:Studied the alteration of the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells before and after PBMC interaction with K562 cells for 24 hours,and also compared the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells after PBMC interaction with two different kind of K562 cells transfected with HLA-B39 and HLA-B51 respectively.Results:After PBMCs were incubated with K562 cells for 24 hours,the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells were both increased.However,after PBMCs were incubated with K562-HLA-B51 cells for 24 hours,the percentage of KIR3DL1+ cells and the percentage of CD16+CD56+ cells were both decreased in comparison with that interaction with K562-HLA-B39 cells.Conclusion:CD16 up-regulation was associated with an up-regulation of inhibitory receptors(KIR3DL1).The interaction between HLA-Bw4 and KIR3DL1 would down-regulate the expression of KIR3DL1.In addition,KIR3DL1 down-regulation was associated with down-regulation of activating receptors(CD16).

Objective To investigate whether MICA gene exon 2,3 and 4 polymorphism is associated with seronegative spondylarthropathies (SpA) or not in Chinese Han population.Methods All 199 B27 positive patients with SpA and 183 randomly ethnically matched healthy controls,and 12 B27 positive controls were enrolled to detect the MICA genotype from its exons 2,3 and 4 by using PCR SSOP method in Shanghai area.Results The MICA007 allele frequency was significantly more in the patient group (18 0%) than in the randomly healthy control group (6 6%) (RR=3 04, P =0 000 045).However,the MICA007 allele frequency was not significantly higher in the B27 positive patient group than in the B27 positive control group.Conclusion The MICA007 allele itself may not be the real disease susceptibility gene involved in the development of ankylosing spondylitis.The increased frequency of MICA007 allele is supposed to be due to a strong linkage disequilibrium between MICA and HLA B genes.

Objective To study the relationship between human leukocyte antigen-DRB1 (HLA-DRB1) allele genes polymorphism and intrahepatic cholestasis of pregnancy (ICP). Methods Forty-two patients with ICP were tested for HLA-DRB1 allele genes polymorphism with the polymerase chain reaction technique and sequence specific oligonucleotide (PCR-SSO) probes hybridization, 56 normal pregnant women as control group were also tested. In addition, the phenotype frequencies of HLA-DRB1 alleles were compared with it′s clinical character in patients with ICP. Results The higher frequencies were observed for alleles DR9, DR12 and DR4 in both groups. DR6 alleles were detected in 14 cases out of 42 patients. Patients with ICP had a significantly higher frequency of the allele DR6 when compared to control group (16.7% vs 3.6%), with a relative risk (RR) as 6.5 (P

Objective:To study the HPLC fingerprint of Xinan capsules from different manufacturers, and establish the chemical pattern recognition method by using principal component analysis and cluster analysis in order to provide reference for the quality con-trol of Xinan capsules. Methods:The HPLC chromatographic column was Agilent ZORBAX SB-C18 (250 mm × 4. 6 mm, 5 μm);the mobile phase was 0.1% formic acid(A)-acetonitrile(B)– tetrahydrofuran(C) with gradient elution, the flow rate was 1.0 ml· min-1;the detection wavelength was 350 nm and the column temperature was 30 ℃. Totally 15 batches of samples were analyzed by the Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprint Similarity (2004A version) and SPSS 19. 0 statisti-cal software. Results:According to the results of cluster analysis and principal component analysis, 10 batches of Xinan capsules were screened out, and the fingerprint common pattern was established. Conclusion:The method is accurate and reliable, and can be used to control the quality of Xinan capsules.

Aim To investigate the effect of matrine on cytoskeletonof K562 cells. Methods Micropipette aspiration technique was adoptedto investigate the viscoelasticity of K562 cells, while the different ex-pression of cytoskeletal protein gene was analyzed by DNA microar-ray. Results In matrine-treated K562 cells, the viscoelastic propertiesKI, K2 and were decreased significantly from 726 ± 215 to 432 ±67,433 ±119 to 242±31, 72±38 to 50±15 respectively, and the geneexpression of prefoldin and ezrin was much stronger than that of controlcells. Conclusion The strueture and function can be changed in ma-trine-treated K562 cells.

Objective To explore the difference of resting-state default-mode network functional connectivity in patients with treatment-resistant depression (TRD) and in healthy subjects. Methods Ten patients with TRD and 12 healthy control subjects underwent 440 s fMRI scans while resting quietly. Functional connectivity analysis was used to isolate the default mode network in each subject. Group maps of the default-mode network were generated and compared between the two groups. A within-group analysis was performed in the depressed group to explore effects of depression refractoriness on network functional connectivity. Results Functional connectivity of both side of middle temporal gyrus, rectal gyrus, precuneus gray matter, left orbital gyrus, right inferior parietal lobule, and post cingulate gyrus in TRD group weakened compared with that of the control subjects. Conclusion There are resting default network connection weakening in multiple brain areas in TRD patients, which may lead to self-control and emotional behavior abnormal in patients.

OBJECTIVETo detect genetic polymorphism in the exons 2 to 4 of the MICA gene in Chinese Han population in Shanghai, Dai population in Yunnan province and Uygur population in Xinjiang province respectively.

METHODSDNA samples from 183 random healthy individuals in Han population, 41 in Dai population and 66 in Uygur population were genotyped by using the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probing (SSOP) method.

RESULTSIn total, 10, 7 and 9 alleles of MICA were observed in Han, Dai and Uygur population respectively. MICA*008 was the most common allele in the population of both Han and Uygur, whereas MICA*010 was the most popular one in Dai population. Han and Dai had a bit similar distribution pattern (Chi-square=12.809 P=0.046), in contrast with Han to Uygur (Chi-square=58.499 P=0) and Dai to Uygur (Chi-square=49.273 P=0).

CONCLUSIONMICA gene displayed different allele distributions in different populations.

Alleles ; China ; Exons ; genetics ; Gene Frequency ; Genotype ; Geography ; Haplotypes ; Histocompatibility Antigens Class I ; genetics ; Humans ; Linkage Disequilibrium ; Polymorphism, Genetic

Objective:To evaluate the effectiveness of enhanced CT texture feature analysis in predicting pseudoprogression in patients with metastatic clear cell renal cell carcinoma (mccRCC) undergoing programmed cell death protein 1 (PD-1) inhibitor therapy.Methods:A cross-sectional study. Data from 32 patients with mccRCC were retrospectively collected who received monotherapy with PD-1 inhibitors after standard treatment failure at Henan Cancer Hospital, from June 2015 to January 2021. Clinical information and enhanced CT images were analyzed to assess target lesion response. The lesions were divided into pseudoprogression and non-pseudoprogression groups. Manual segmentation of target lesions was performed using ITK-Snap software on baseline enhanced CT, and texture analysis was conducted using A.K. software to extract feature parameters. Differences in texture features between the pseudoprogression and non-pseudoprogression groups were analyzed using univariate and multivariate logistic regression. A predictive model for pseudoprogression was constructed, and its performance was evaluated using ROC curve analysis.Results:A total of 32 patients with 89 lesions were included in the study. Statistical analysis revealed significant differences in seven texture features between the pseudoprogression and non-pseudoprogression groups. These features included“original_ngtdm_Strength”(0.49 vs. -0.61, P=0.006), “wavelet-HLH_glszm_ZonePercentage”(0.67 vs. -0.22, P=0.024),“wavelet-LHL_ngtdm_Strength”(1.20 vs. -0.51, P=0.002), “wavelet-HLL_gldm_LargeDependenceEmphasis”(-0.84 vs. 0.19, P=0.002), “wavelet-HLH_glcm_Id” (-0.30 vs. 0.43, P=0.037),“wavelet- HLH_glrlm_RunPercentage”(0.45 vs. -0.01, P=0.032),“wavelet-LHH_firstorder_Skewness”(0.25 vs. -0.27, P=0.011). Based on these features, a pseudoprogression prediction model was developed with a P-value of 0.000 2 and an odds ratio of 0.045 (95% CI 0.009-0.227). The model exhibited a high predictive performance with an AUC of 0.907 (95% CI 0.817-0.997) according to ROC curve analysis. Conclusions:Enhanced CT texture feature analysis shows promise in predicting lesion pseudoprogression in patients with metastatic ccRCC undergoing PD-1 inhibitor therapy. The developed predictive model based on texture features demonstrates good performance and may assist in evaluating treatment response in these patients.

Objective:To investigate the ability of the anti-PD-1(scFv)/hIL-15 fusion protein(PD-15) to specifically bind to PD-1 in vitro and the effect of the combination of PD-15 with GF-hIL-15Ra-K562(G15Ra-K562) feeder cells to expand NK/T cells. Methods:Overlap PCR was used to construct G15Ra expression vector. pMXs-G15Ra-IP was transfected into K562 by electroporation. G15Ra-K562 feeder cell lines were obtained by limiting dilution method. pUC57-PD-15 was constructed by digestion and ligation. Lipofectamine? 2000 was used to transiently transfect pUC57-PD-15 into HEK293T cells and the conditioned medium containing PD-15 fusion protein was obtained. Density gradient centrifugation was used to obtain human peripheral blood mononuclear lymphocytes(PBMC), and CFSE staining was used to mark active proliferating cells. Flow cytometry was used to detect the ability of PD-15 to specifically bind to PD-1 and its effect on the proliferation of human PBMC and the proportion of different subpopulations of lymphocytes.Results:The feeder cells G15Ra-K562 with high expression of fusion protein G15Ra was successfully constructed. The addition of hIL-15 can increase the ability of G15Ra-K562 to expand human PBMC by more than 5 times( P<0.05). PD-15 fusion protein has PD-1 specific binding ability( P<0.001), combined with G15Ra-K562 can efficiently expand human peripheral blood-derived NK/T cells in vitro( P<0.05). The cells expanded by PD-15 and G15Ra-K562 are mainly natural killing cell, CD8 + T and CD4 + T cells. Conclusions:The PD-15 fusion protein can specifically target the PD-1 molecule and has a strong human peripheral blood-derived NK/T cell expansion ability when combined with G15Ra-K562 feeder cells. These results shed light on selective expansion of PD-1 + lymphocytes in vitro.