Objective To investigate the relationship between polymorphism of resistin(RETN)gene and metabolic associated fatty liver disease(MAFLD)in type 2 diabetes mellitus(T2DM)patients in middle and high altitude areas.Methods A total of 400 patients with T2DM in Qinghai area were recruited and divided into simple T2DM group(T2DM,n=200)and T2DM combined with MAFLD group(T2DM+ MAFLD,n=200)according to liver ultrasonography.Healthy individuals confirmed by physical examination were selected as the normal control group(NC,n=180).Plasma resistin levels were measured by ELISA.The polymorphism of RETN-420C/G and +299G/A genes were detected by PCR sequencing.Results By comparing the polymorphism of RETN-420C/G gene in each group,it was found that the frequencies of G/G genotype and G allele frequency in T2DM+MAFLD group were higher than those in NC group and T2DM group(P<0.05),while the frequencies of C/C genotype and C allele frequency were lower than those in NC group and T2DM group(P<0.05).The risk of MAFLD increased by 1.571,2.126 and 1.537 times respectively in T2DM patients with C/G,G/G genotype and G allele.Logistic regression analysis showed that G/G genotype was a risk factor for MAFLD in T2DM patients.By comparing the polymorphism of RETN+299G/A gene in each group,it was found that A allele frequency in T2DM+MAFLD group was higher than that in NC group and T2DM group,while G allele frequency was lower than that in NC group and T2DM group(P<0.05).The allele A increased the risk of MAFLD in T2DM patients by 1.432 times compared to allele G.Conclusion RETN gene-420C/G locus G/G genotype increases the risk of T2DM combined with MAFLD in middle and high altitudeareas.

AIM: To observe the clinical efficacy of multi -glycoside of tripterygium wilfordii (GTW) on diabetic nephropathy. METHODS: Fifty-one patients with diabetic kidney disease (DKD) with a history of GTW dosing admitted to the outpatient clinic of Yijishan Hospital affiliated to Wannan Medical College from June 2019 to October 2022 were selected as study subjects, and were followed up regularly to observe the changes in laboratory indexes before and after GTW dosing and adverse drug reactions after 6 months of treatment. The t-test, Mann-Whitney U-test or χ

Objective:To study the effect of thioredoxin domain containing protein 5 (TXNDC5)-peroxiredoxin 2 (Prx2) on the drug resistance of prostate cancer cells.Methods:Prostate cancer PC3 cells were cultured in vitro, treated with the chemotherapy drug cyclophosphamide (5, 10, 15 μmol/L) for 24 hours, and PC3 cells without any treatment was served as the control group. The expression levels of TXNDC5 in PC3 cells were detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting. PC3 cells with TXNDC5 knocking down were exposed by cyclophosphamide and CCK-8 was used to detect the cell viability of siTXNDC5 group and siNC group. The content of reactive oxygen free radicals was determined by reactive oxygen detection kit. PC3 cells and its parental cyclophosphamide-resistant ones with TXNDC5 knocking down were treated by 10 μmol/L cyclophosphamide and subjected for CCK8 assay. The expression of Prx2 in PC3 cells was detected by Western blotting after TXNDC5 was silenced. Prx2 expression was silenced in PC3 cells overexpressing TXNDC5, and cell viability and reactive oxygen free radical content were detected in Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group. Results:Compared with the control group, cyclophosphamide treatment significantly increased the expression of TXNDC5 at mRNA and protein levels in PC3 cells. After PC3 cells were treated with cyclophosphamide (10, 15 μmol/L) for 12 h, compared with the siNC group, the cell viability in the siTXNDC5 group was significantly suppressed (0.44±0.08 vs. 0.74±0.10, t=3.647, P=0.031; 0.30±0.04 vs. 0.53±0.06, t=6.115, P=0.006). When PC3 cells were treated with 10 μmol/L cyclophosphamide for 6 and 12 h, compared with the siNC group, the production of reactive oxygen free radicals in the siTXNDC5 group was significantly increased (2.68±0.19 vs. 1.58±0.26, t=-6.027, P=0.005; 4.56±0.37 vs. 2.73±0.26, t=-6.995, P=0.003). When PC3 cells and its cyclophosphamide-resistant ones were treated with 10 μmol/L cyclophosphamide for 12 h, compared with the siNC group, the cell viability was significantly inhibited in the siTXNDC5 group. Western blotting analysis showed that the expression of Prx2 was significantly reduced when TXNDC5 was silenced. Silencing Prx2 could significantly attenuate the increase of cell viability and the decrease of reactive oxygen content resulting from TXNDC5 overexpression. PC3 cells were treated with 10 μmol/L cyclophosphamide for 12 h, and the cell viabilities of the Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group were 0.52±0.07, 0.69±0.03, 0.56±0.05, 0.43±0.05, 0.58±0.07, respectively, and there was a statistically significant difference ( F=8.868, P=0.003). Furthermore, the cell viability in the pcTXNDC5+ siPrx2 group decreased significantly when compared to that of the pcTXNDC5 group ( P=0.045). The contents of reactive oxygen free radicals in the above 5 groups were 3.26±0.46, 2.09±0.49, 3.16±0.38, 4.62±0.26, 2.87±0.36, respectively, and there was a statistically significant difference ( F=16.037, P<0.001). The content of reactive oxygen radicals in the pcTXNDC5+ siPrx2 group was higher than that of the pcTXNDC5 group ( P=0.036). Conclusion:TXNDC5 can reduce the level of reactive oxygen free radicals in prostate cancer cells by regulating the expression of Prx2, so as to promote the drug resistance of prostate cancer cells.

Objective To improve the efficiency of result reporting and ensure the accuracy of the results by establishing autoverification system in Clinical Chemistry and Immunology Laboratory.Methods The study followed the requirements of the Clinical Laboratory Standards Institute(CLSI)AUTO-10A and ISO 15189:2012.In addition,seven categories of verification rules were encoded using the autoverification function of the CentraLink?Data Management System on the Aptio?Automation platform.These rules included Clinical Diagnostic Standard(CS), Sample Status(SS), Quality Control Severity(QS), Instrument Error Flags Severity(IS), Normal Severity(NS), Delta Check Severity(DS), and Logical Assessment Standard(LS).Various modules of Aptio Automation,laboratory information system(LIS)and hospital information system(HIS)were integrated using the CentraLink system to establish the autoverification system.Results The autoverification system was set up and tested from August 2015 to April 2016.In total, the system ran 4 496 425 tests on 366 180 chemistry specimens.The overall autoverification rate for tests performed increased from 53.4% to 87.0%.Glucose had the highest rate (98.3%)while CKMB had the lowest rate(63.6%).Average TAT for result verification decreased by 97.7%,from 46.3 minutes to 3.7 minutes.The system ran 410,040 tests on 160 119 chemiluminescence specimens.The autoverification rate for tests performed increased from 40.2%to 89%.C-P had the highest rate(98.4%)while A-TPO had the lowest rate(58.7%).Average TAT for result verification decreased by 77.4%,from 14.6 minutes to 3.3 minutes.From May 2016 to January 2017(when autoverification was employed),compared with the same period in 2014(when manual verification was employed),the following changes were observed with no increase in staff capacity:a)Volume of routine chemistry tests increased by 46.4%,and median TAT for tests decreased by 41.9%, from 118 minutes to 83 minutes; b)Volume of chemiluminescence tests increased by 24.5%and median median TAT for tests decreased by 52.4%, from 131 minutes to 86 minutes;c)Median TAT for critical values decreased by 50.5%; d)Rates of tests that did not go through autoverification were 88.2% for NS,6.05% for SS, 2.40% for DS,2.00% for LS, 0.97%for IS,and 0.43% for CS; e)Rates of abnormal specimen status identified by Aptio Automation were 7.13‰for jaundice,5.39‰ for blood lipids,2.20‰ for hemolysis,0.17‰ for barcode error, and 0.15‰ for insufficiency;f)Error rate decreased to 0.00%;and g)staff satisfaction increased from 85%to 100%.Conclusion Autoverification of results by using the CentraLink Data Management System can achieve quality control over the entire process of clinical laboratory testing, ensure accuracy of test results, improve work efficiency, decrease TAT, minimize the error rate, avoid skill variation of staff, reduce the pressure of performing manual verification,and improve medical security.

Objective To analyze the causes of hypoglycemia coma in emergency diabetes patients and its corresponding nursing strategies.Methods The reasons of hypoglycemia coma was retrospectively analyzed in our hospital and the risk factors of hypoglycemic coma were analyzed by multivariate logistic regression,and the corresponding nursing strategies were summarized.Results Of the 767 critically ill patients,100 (13.04％) had hypoglycemia,among whom 27 patients (3.52％) had severe hypoglycemia.Multivariate logistic regression analysis showed that patients with type 2 diabetes history,hypoglycemic drugs therapy,hypoproteinemia,renal insufficiency,hemodialysis,malnutrition were risk factors for hypoglycemic coma.While normal hemoglobin was a protective factor for hypoglycemic coma.Conclusion Hypoglycemia is the result of multiple factors in emergency patients.Reasonable nursing measures can effectively prevent the occurrence of hypoglycemic coma in emergency patients.

Objective To analyze the causes of hypoglycemia coma in emergency diabetes patients and its corresponding nursing strategies.Methods The reasons of hypoglycemia coma was retrospectively analyzed in our hospital and the risk factors of hypoglycemic coma were analyzed by multivariate logistic regression,and the corresponding nursing strategies were summarized.Results Of the 767 critically ill patients,100 (13.04％) had hypoglycemia,among whom 27 patients (3.52％) had severe hypoglycemia.Multivariate logistic regression analysis showed that patients with type 2 diabetes history,hypoglycemic drugs therapy,hypoproteinemia,renal insufficiency,hemodialysis,malnutrition were risk factors for hypoglycemic coma.While normal hemoglobin was a protective factor for hypoglycemic coma.Conclusion Hypoglycemia is the result of multiple factors in emergency patients.Reasonable nursing measures can effectively prevent the occurrence of hypoglycemic coma in emergency patients.

Objective To investigate the precision, trueness, and accuracy of self-developed detection system in clinical chemistry.Methods This was a methodological evaluation.Take serum creatine kinase( CK) for instance, 6 serum sampleswith different leves ( on the upper or lower limit of the reference range or close to the medicine decide levels ( MDLs) , were collected for within-run precision( repeatability) and within-laboratory precision ( intermediate precision ) experiments.5 proficiency testing ( PT ) samples, 5 samples assigned value by reference method, and 40 fresh-frozen serums were measured and compared with reference method for trueness verification.Drawing method evaluation decision chart, calculating total errors and sigma level evaluation experiment based on the CV, bias, and allowed total errors(TEa)were used to evaluate the accuracy performance.The precision, trueness, and accuracy were compared with the quality indicators.Results The within-run precision and within-laboratory precision were less than the highest requirement of Chinese industrial standard.The mean bias was -8.96%, didn′t reachthe required standard (5.5%).After taking corrective actions, all samples but one ( -5.8%) met the required standard. Compared with the reference method, the mean bias on the MDLs was less than TEa.The performance points of the method evaluation decision chart indicated excellence performance.The total errors on MDLs were 14.2%, 10.4% and 7.6%, less than 15%.The sigma levels on MDLs were 5.9, 7.5 and 15, also achieved excellent level. Conclusions The precision, trueness, and accuracy performance of CK measured by self-developed detection system achieved excellent level of the Chinese industry standard, and the same results were found from different evaluation methods.

Objective To estimate the methodology for evaluating the analytical measurement range ( AMR) and the clinically reportable range ( CRR) in lab-developed system in clinical chemistry .Methods Method evaluation .Take serum CK for instance , a series of samples were prepared from both a specimen with a high concentration of the analyte of interest and a specimen with a low concentration for the following assays.Average slope method , linear dilution recovery method and the method recommended by Clinical and Laboratory Standards Institute ( CLSI ) EP6-A were used to established AMR in the lab-developed clinical chemistry system.Based on the maximum valid dilution of the specimen and the results of AMR , CRR were determined.One-half of the total error allowance ( TEa) of Chinese health standard was set up as allowance error (7.5%).Results X-Y scatter plot was made by assigning sample numbers to the horizontal axis and actual measured values to the vertical axis , which determined the upper limit of AMR was approximate 1 651 U/L.The results analyzed by average slope method indicated that the linear correlation between expected values and actual measured values was determined , the correlation (r), the intercept (a) and the slope (b) met the linear standard, and AMR was 5-1 699 U/L.The results analyzed by EP6-A indicated that the best fitting curve was obtained by using cubic polynomial method , and the linearity deviation of the minimum concentration was -77.1%, which exceeded one-half of TEa.Followed by the deletion of the maximum concentration , the resumed experiment was done .The results showed that the nonlinear coefficient c of quadratic polynomial and the nonlinear coefficient c and d of cubic polynomial have no significant difference to 0, and AMR was 7.5-1 458.0 U/L.By linear dilution recovery method , the linear correlation between expected values and actual measured values was determined , the correlation ( r) , the intercept ( a) and the slope ( b) met the linear standard , the recovery rates was between 100.0%and 104.8%, and AMR is 5 -1 699 U/L.The CRR was determined to be 5 -33 880 U/L, which met the standard of TEa . Conclusions Average slope method, linear dilution recovery method and EP 6-A method were all used to established AMR in lab-developed clinical chemistry system .Without complicated statistical analysis , linear dilution recovery method was suitable for clinical use .The linearity deviation of the minimum concentration analyzed by EP6-A did not meet the standard of the quality objective system , suggesting defects in the statistical analysis of the results .CRR was feasibly determined by using linear dilution recovery method align with AMR.

Objective To investigate the association between gene polymorphism of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and Graves ophthalmopathy (GO) in Qinghai Han population.Methods Ninety cases of Graves disease were selected from June 2011 to February 2014 in The People's Hospital of Qinghai Province,and the 90 patients were divided into two subgroups according to GO (49 cases) and GD without GO(41 cases).Then the genotype and allele of CTLA-4 exon 1 (+ 49A/G) were detected in surum by the method of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).Results The distribution of CTLA-4 exon 1 (+ 49 A/G) genotype frequencies (AA,AG,GG) was not different between GO and GD without GO subgroups [4.1％ (2/49) to 7.3％ (3/41),44.9％ (22/49) to 61.0％ (25/41),51.0％ (25/49) to 31.7％ (13/41),Fisher exact probability,P =0.180 ＞ 0.05]; the distribution of CTLA-4 exon 1 (+ 49A/G) allele frequencies (A,G) was not different between GO and GD without GO subgroups [26.5％ (26/98) to 37.8％ (31/82),73.5％ (72/98) to 62.2％ (51/ 82),x2 =2.622,P＞ 0.05].Conclusion CTLA-4 gene exon 1 (+ 49A/G) may not be a candidate susceptibility gene for Qinghai Han GO.

BACKGROUND:Recent studies have found that bone marrow mesenchymal stem cels that culturedin vitro for a long time can naturaly differentiate into neural stem cels, which then differentiate into neurons and glial cels, thereby providing a new therapeutic thinking for Parkinson’s disease, sequela of cerebral infarction, cerebelar atrophy and brain dysplasia.
OBJECTIVE:To discuss the influence of neural stem cel transplantation on neurologic function of rats with cerebral hemorrhage at recovery stage and the relevant mechanism of action.
METHODS: Sixty male Sprague-Dawley rats were randomly divided into normal group (n=18), cerebral hemorrhage group (n=21) and transplantation group (n=21). Cerebral hemorrhage models were established in the latter two groups using VII type colagen enzyme induction method. At 21 days of modeling, rats in the transplantation group were injected neural stem cels via the tail vein, and those in the other two groups received the same volume of normal saline. At 7, 14, 21 days after cel transplantation, modified adhesive removal test (MST) was employed to evaluate the neurologic function of rats, and then the rats were kiled. RT-PCR was used to detect angiopoietin-1 mRNA expression in the bleeding tissues, and western blot assay was employed to measure tyrosine kinase receptor-2 protein expression.
RESULTS AND CONCLUSION:Compared with the normal group, the MST scores in the cerebral hemorrhage group and transplantation group were significantly decreased (P< 0.05). From the 7th day after transplantation, MST scores in the transplantation group were significantly higher than those in the cerebral hemorrhage group (P < 0.05). At 7, 14, 21 days after transplantation, expressions of angiopoietin-1 mRNA and tyrosine kinase receptor-2 protein were ranked as folows: transplantation group > cerebral hemorrhage group > normal group, and there was a significant difference among the three groups (P< 0.05). These findings indicate that neural stem cel transplantation can effectively promote the neurologic recovery of rats with cerebral hemorrhage at recovery stage, and the concrete mechanism may be related to the increase of angiopoietin-1 mRNA and tyrosine kinase receptor-2 protein in the bleeding tissues.