The up-regulation mechanism of membrane type I matrix metalloproteinase (MT1-MMP) in macrophages stimulated by silica in vitro and the contribution of early growth response 1 (Egr-1) transcription factor in the gene expression pathway were investigated. Macrophages stimulated by silica were treated with Egr-1 antibody or Egr-1 decoy oligodeoxynucleotides (ODN). The levels of MT1-MMP proteins were determined by Western blot and the expression of MT1-MMP mRNAs was detected by RT-PCR. The results showed as compared with control macrophages, silica-stimulated group showed up-regulated gene expression of MT1-MMP via Egr-1 (P<0.01). Compared with silica-stimulated macrophages untreated with antibody, the cells treated with 5 microg/mL Egr-1 antibody were associated with reduced expression of MT1-MMP protein (P<0.01) and mRNA (P<0.01). Compared with silica-stimulated untransfected group, the Egr-1 "decoy" ODN group was associated with reduction in the expression of MT1-MMP protein and mRNA (P<0.01). It was concluded gene expression of MT1-MMP which may play a critical role in silicosis was up-regulated by silica in macrophages. Egr-1 participated in the expression of MT1-MMP and positively regulated the expression. Both Egr-1 antibody and Egr-1 decoy ODN suppressed the expression of MT1-MMP through the Egr-1 pathway and may become a potential therapeutic tool in the management of silicosis in the future.

Objective: To investigate the epidemiological characteristics of varicella in Yandu District, Yancheng City, Jiangsu Province from 2005 to 2021, so as to provide insights into the improvements of the varicella control strategy. Methods: The data pertaining to cases in Yandu District from 2005 to 2021 were captured from the National Notifiable Diseases Network Reporting System, and the coverage of varicella vaccination in Yandu District from 2016 to 2021 was captured from the Jiangsu Provincial Immunization Planning System. The trends in incidence of varicella were analyzed using annual percent change (APC), the temporal, population and spatial distributions of varicella cases were descriptively analyzed, and the correlation between the cumulative first-dose varicella vaccination rate and incidence of varicella was examined among birth cohorts in Yandu District from 2005 to 2021. Results: Totally 6 144 varicella cases were reported in Yandu District from 2005 to 2021, with annual mean reported incidence of 50.05/105. The incidence of varicella appeared a tendency towards a rise in Yandu District from 2012 to 2018, with APC of 77.97% (t=11.702, P<0.001), and the incidence appeared a tendency towards a decline from 2018 to 2021, with APC of -41.93% (t=-6.911, P<0.001). The incidence of varicella peaked during the period between November and January of the following year (45.95%), and during the period between May and June (20.35%). The mean incidence of varicella was higher among men than among women (54.72/105 vs. 45.00/105; χ2=57.858, P<0.001). The highest incidence of varicella was found among cases at ages of 6 to 9 years (543.78/105), and varicella predominantly occurred among primary school students (38.40%) and pre-school children (31.85%), with high annual mean incidence in the eastern urban areas (74.27/105).The cumulative first-dose varicella vaccination rate negatively correlated with the incidence of varicella among the birth cohorts in Yandu District in 2019 (r=-0.501, P<0.001), 2020 (r=-0.646, P<0.001) and 2021 (r=-0.602, P<0.001). Conclusions The incidence of varicella appeared a tendency towards a rise followed by a reduction in Yandu District from 2005 to 2021, and the incidence of varicella peaked in winter and at the turn from spring to summer, with varicella predominantly found among primary school students and preschool children.

Objective: To understand the survival status and influencing factors of HIV/AIDS patients with highly active antiretroviral therapy ( HAART ) among drug users in Yili Prefecture, Xinjiang from 2005 to 2019, so as to provide references for reducing AIDS mortality. Methods : The demographic information, clinical stage, baseline CD4+T lymphocyte ( CD4 ) level and treatment status of HIV/AIDS patients with HAART in Yili Prefecture from 2005 to 2019 were collected through AIDS Antiretroviral Therapy Information System. The survival rate was calculated by the life table method. The influencing factors for survival time were analyzed by Cox proportional hazard regression model. Results: Totally 1 935 patients were recruited, the median age receiving HAART was 37 years old and the median CD4 counts was 293/μL. The cumulative survival rates at 1, 5, 7 and 10 years were 97%, 78%, 73%, and 66%, respectively. The multivariate Cox proportional hazards regression analysis showed that the patients with body mass index of 18.5-<28.0 kg/m2 ( HR: 0.391-0.656, 95%CI: 0.234-0.958 ), baseline CD4>200/μL ( HR: 0.354-0.667, 95%CI: 0.232-0.841 ) , or missed medication in the last 7 days ( HR=0.009, 95%CI: 0.001-0.061 ) had lower risk of death; the patients with WHO clinical stage of Ⅱ-Ⅳ ( HR: 1.479-2.311, 95%CI: 1.004-3.288 ) or treatment delay ≥1 years ( HR: 1.287-1.388, 95%CI: 1.029-1.826 ) had higher risk of death. Conclusions The 5-year cumulative survival rate of HIV/AIDS patients with HAART in Yili Prefecture is 78%. Body mass index, baseline CD4 level, WHO clinical stage, treatment delay and missed medication in last 7 days were the influencing factors for survival time.

OBJECTIVETo explore the expansion method of high purity NK cells from human peripheral blood and explore the changes in biological functions of NK cells after ex vivo expansion.

METHODSNK cells were isolated from peripheral blood mononuclear cells (PBMNCs) by using miniMACS (Magnetic cell-selection) and NK Cell Isolation Kit II, and cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of interleukin (IL)-2 and/or IL-12, IL-15 for 15 days. Cultures were semi-exchanged with fresh media and cytokines every 3 days. Evaluation for cell expansion, phenotype, perforin and granzyme B mRNA expressions, and IFN-gamma secretion before and after the culture period.

RESULTSCD3(-) CD56(+) cells concentration increased from (11.2 +/- 5.2)% to (94.2 +/- 3.5)%. In group IL-2 + IL-15 and IL-2 + IL-15 + IL-12 group, cells were expanded 50.5 +/- 4.3 and 52.3 +/- 6.7 - fold, respectively, being significantly higher than that in other three groups [(15.4 +/- 1.1 fold in IL-2 group, 19.9 +/- 3.9 fold in IL-2 + IL-12 group, 6.1 +/- 1.0 fold in control group)] (P<0.01), but no significant difference between each other (P>0.05). The purity of CD3(-) CD56(+) NK cells was over 94% in all groups except the control. The perforin and granzyme B mRNA expressions of expanded NK cells in four experimental groups were significantly higher than those of before expansion (P<0.01) and the expressions in IL-2 + IL-15 and in IL-2 + IL-12 + IL-15 group were significant higher than in other three groups (P<0.01) while no significant difference between each other (P>0.05). IFN-gamma levels in the supernatants of four experiment groups were significantly higher than that in control group (P<0.01) and its levels order was IL-2 + IL-15 + IL-12 group > IL-2 + IL-12 group > IL-2 + IL-15 group > IL-2 group (P<0.01).

CONCLUSIONHigh purity NK cells isolated by negative selection using miniMACS can be efficiently expanded with IL-2 + IL-15, and their biological functions were enhanced.

Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; pharmacology ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukins ; pharmacology ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; metabolism ; Perforin ; metabolism

Objective Brucine and strychnine monomer reference substance as extremely toxic substance had potential threat during transportation and utilization. In this study we investigated the homogeneity, stability and assignment accuracy of the mixture reference solution of strychnine and brucine, so as to provide reference for the quality control of extremely toxic chemical reference substances for Chinese medicine. Methods Following the assay in Chinese Pharmacopoeia volume I (2015), we prepared the mixture reference solution of brucine and strychnine, and investigated the solvents and the concentration of mixutre reference solution. The stability test lasted for 12 months. F-test was used for heterogeneity assay. Three researchers were involved for collaboration. Results Methanol and chloroform solution were selected as the solvents for the stability test. Results showed the difference was not statistically significant among various mixture solutions. The results of value assignment were 0.14 mg/mL for strychnine (sR = 0.5％)and 0.10 mg/mL for brucine (sR = 1.0％). The stability of mixture solution were better under the conditions of methanol solution at 4 ℃ or -20 ℃. Conclusion The results provide a possible way to develop the mixture solution in place of the monomer reference, and the mixture reference solution is expected for the quality control in the slices of Semen Strychni and its compound preparations.

Objective To investigate DNA methylation markers in the whole blood of patients with systemic lupus erythematosus(SLE),in hope to facilitate the evaluation of SLE severity.Methods Whole blood samples were obtained from 58 patients with SLE(including 14 cases of severe SLE,25 moderate SLE,19 inactive SLE)and 50 healthy controls.Bisulphite sequencing was performed to determine the methylation status of interleukin-2 common receptor gamma chain(IL-2RG)promoter region,and real-time reverse transcriptionPCR to quantify the expression level of IL-2RG mRNA,in these subjects.Results The methylation level of IL2RG promoter region was 0.217 ± 0.140,0.325 ± 0.230,0.342 ± 0.085 and 0.175 ± 0.036 in the patients withsevere,moderate and inactive SLE and healthy controls,respectively.A significant increase was observed in the methylation level of IL-2RG promoter region in the patients with inactive SLE compared with the patients with severe SLE and healthy controls(both P ＜ 0.01),and in the patients with SLE compared with the healthy controls(0.263 ± 0.047 vs.0.175 ± 0.036,P ＜ 0.05).The expression level of IL-2RG mRNA was significantly lower in the patients with SLE than in the healthy controls(2.550 ± 0.823 vs.4.293 ± 1.283,P ＜ 0.05).A negative correlation was observed between the expression level of IL-2RG mRNA and methylation level of IL2RG promoter region in 20 patients with SLE(r =-0.44,P ＜ 0.05).Conclusion The methylation status of IL2RG promoter region is statistically higher in patients with SLE than in healthy controls,and significantly different between patients with active SLE and those with stable SLE.

OBJECTIVETo study the effects of Qiwei Granule (QWG) on the protein and mRNA expressions of renal tissue transforming growth factor beta1 (TGF-beta1) in KK-Ay mice with spontaneous type 2 diabetes mellitus (T2DM).

METHODSSpontaneous T2DM KK-Ay mice model was adopted. Forty-five male mice were randomly divided into three groups, i. e., the model group, the Chinese medicine group, and the Western medicine group, 15 in each group. Fifteen male C57BL/6J mice were set up as the normal control group. The mice in the Chinese medicine group and the Western medicine group were administered intragastrically with QWG (at the daily dose of 20 g/kg) and valsartan (at the daily dose of 10 mg/kg), and the treatment lasted for 12 successive weeks. The pathological changes of the kidney were observed using HE staining, PAS, and Masson staining. The protein and mRNA expressions of TGF-beta1, were detected using immunohistochemical method and Real-time fluorescent quantitative PCR.

RESULTSThe renal pathological changes of mice in the model group showed hypertrophic glomeruli, widened mesenteric matrix, increased mesangial cells, vacuolar renal tubular epithelial cells, tubular ectasia, and foci atrophy. Necrosis was occasionally seen. More protein cast, mesenchymal infiltration of inflammatory cells, and interstitial fibrosis could be seen. The protein and mRNA expressions of TGF-beta1 increased more in the model group than in the normal control group. After treatment by QWG and valsartan, the renal pathological changes were obviously alleviated, and the protein and mRNA expressions of TGF-beta1 were obviously lowered (P<0.05).

CONCLUSIONBy inhibiting the protein and mRNA expressions of TGF-beta1, QWG could play a role in preventing and curing diabetic nephropathy.

Animals ; Diabetes Mellitus, Type 2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Kidney ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Select patients diagnosed of hypertension in the first place in 16 hospitals and the patients were described by association rules analysis and distribution for the analysis in the study,in order to understand the information of diagnosis and treatment in hypertension patients in real world. The information include age, gender, admission condition, inpatient department, hospitalization expenses, western medicine comorbidities, the traditional Chinese medicine (TCM) syndrome and the medicine. The average age of patients was 64 years in the study. The proportion of men to women about 1. 10: 1. Admission condition is critical for about 10.12% of the total and more concentrated in the cardiovascular department. Hospital stay ranged from 8 to 14 d. Inpatients with medical insurance is 62.93%. Total hospitalization cost distribution most is the 5 000-10 000 RMB. Hypertension complicated with coronary heart disease, blood lipoprotein disorder disease; phlegm and blood stasis and Yin deficiency of liver and kidney are the most in TCM syndrome type. Promoting blood circulation and removing blood stasis is the major in Chinese medicine treatment. Western medicine treatment basically conforms to the guidelines. The most antihypertensive drugs is calcium antagonistst, accounting for about 81.2% of the total number. Class ACEI, class ARB, beta blockers, diuretics class are accounted for 43.0%, 43.4%, 42.4%, 42.4%, fixed compound is only 2 393, accounting for about 8%. the total frequency of five classes of antihypertensive drugs is 78 206 times. The principal conclusions of this analysis are as follows: the elderly people is the most in hypertension, more men than women; medical insurance is the majority type in hospitalization cost; nearly half of the population is combined with coronary heart disease; phlegm and blood stasis and yin deficiency of liver and kidney are the most in TCM syndrome type; western medicine treatment basically conforms to the guidelines. Combination therapy is more, less fixed compound preparation in western medicine. Chinese medicine is mainly to promoting blood circulation. Suggesting that the clinical treatment in strict blood pressure and at the same time, pay attention to combination therapy and the use of fixed compound preparation,it's useful to control the blood pressure better. Combined with traditional Chinese medicine and resolving phlegm and removing blood stasis, reduce the target organ damage, reduce complications, the prevention and control work of hypertension in China still faces enormous challenges at present. The result provides reference information for clinical medicine and medical research.
Adolescent ; Adult ; Aged ; Antihypertensive Agents ; therapeutic use ; Blood Pressure ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension ; complications ; drug therapy ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Treatment Outcome ; Young Adult

Objective :To explore change of plasma interleukin 15 (IL-15) level and its correlation with N terminal pro B-type natriuretic peptide (NT-proBNP).Methods :A total of 205 patients diagnosed as CHF were sequentially en-rolled as CHF group ,and another 52 healthy adults undergoing physical examination simultaneously were regarded as healthy control group .According to NYHA cardiac function class ,CHF group was further divided into class II group (n＝55) ,class III group (n＝87) and class IV group (n＝63).Plasma IL-15 level was measured and compared among all groups ,and correlation among plasma level of NT-proBNP ,left ventricular ejection fraction (LVEF) and plasma IL-15 level were analyzed in CHF patients .Results : Compared with healthy control group ,there was signif-icant rise in plasma NT-proBNP level [ (71 ± 33) pg/ml vs.(5884 ± 1379) pg/ml] ,and significant reduction in LVEF [(57 ± 7)% vs.(31 ± 8)% ] in CHF group ,P＝0.001 both .Plasma IL-15 level of CHF group was significant-ly higher than that of healthy control group [(18.35 ± 6.42) ng/L vs.(7.78 ± 3.11) ng/L] , P＝0.001 ;compared with class II group ,there was significant rise in plasma IL-15 level [ (10.24 ± 3.61) ng/L vs.(20.17 ± 6.35) ng/L vs.(21.94 ± 7.32) ng/L] in class III and IV group , P＝0.001 both ,and there was no significant difference be-tween class III and class IV group (P＝0.187).Pearson correlation analysis indicated that plasma IL-15 level was significant positively correlated with plasma NT-proBNP level ( r＝ 0.172 , P＝ 0.038).Conclusion :Plasma IL-15 level of CHF patients is significantly higher than that of healthy subjects ,and significant positively correlated with NT-proBNP level ,and it has certain relationship with heart failure severity ,so it may become a new marker for pre-dicting CHF .

Human glutamate decarboxylase 65(hGAD65) is an enzyme that catalyzes the transformation of L-glutamic acid into ?-aminobutyric acid.It has been found that Type 1 diabetes mellitus(T1DM)is an autoimmune disease,in which pancreatic islet ?-cells are destroyed due to immune response mediated by autoantigen.hGAD65 is considered as a key autoantigen of the autoimmune response,so anti-hGAD65 antibody(hGAD65-Ab) is the most effective and specific immune marker for T1DM diagnosis,and hGAD65 can be used to detect hGAD65-Ab in serum of T1DM patients.The hGAD65 gene was cloned into pET32a(+),then the recombinant plasmid with hGAD65 was transformed into E.coli BL21(DE3) and expressed by IPTG induction.The fusion protein containing thioredox,hexahistidine and hGAD65(Trx-hGAD65) was mostly insoluble,but the band of soluble Trx-hGAD65 could also be detected by SDS-PAGE,and it was a great improvement compared with the results reported.Trx-hGAD65 was isolated from lysate and purified by immobilized metal ion affinity chromatography(IMAC).After enterokinase digestion and IMAC purification,hGAD65 with high purity was obtained.Detection of thin-layer chromatography(TLC) showed that both Trx-hGAD65 and hGAD65 had enzymatic activity,whereas Trx-hGAD65 had better stability.Furthermore,it was confirmed that Trx-hGAD65 was able to conjugate with hGAD65-Ab in the serum of T1DM patients by ELISA assay.In conclusion,Trx-hGAD65 instead of hGAD65 can be used for T1DM diagnosis,and its application in prophylaxis and therapy of T1DM is expectable.