Objective To explore the 5,10-methylenetetrahydrofolate reductase( MTHFR) and 5-methyltet-rahydrofolate-homocysteine methyltransferase reductase( MTRR) gene polymorphisms among the Han women in Laiwu City.Methods A total of 559 Han women were recruited.And their oral epithelial cells were collected to extract genome DNA in order to detect gene polymorphisms of MTHFR and MTRR using fluorescence quantitative PCR.Then the results were compared with those in other cities in China.Results The frequency of MTHFR 677CC,677CT and 677TT of Han women in Laiwu city was 14.3%,46.7%and 38.1%,respectively.The frequency of MTHFR 677TT among Laiwu women was significantly different to those of Zhenjiang, Wuhan, Kunming, Deyang, Huizhou, Qionghai (P<0.05).The frequency of AA,AC,CC gene type on MTHFR A1298C was 78.2%,19.7% and 2.1%,respec-tively, the frequency was significantly different to those of Zhenjiang, Wuhan, Kunming, Deyang, Huizhou, Qionghai (P<0.05).The frequency of AA,AG,GG gene type on MTRR A66G was 53.3%,38.8%and 7.9%,respectively. The frequency of MTRR 66GG was significantly different to that of Qionghai(P<0.05).Conclusion The MTHFR, MTRR polymorphism distribution of Han women in Laiwu City has the characteristic of region specificity,respectively.

Objective·To directly detect the bacteria in positive blood culture specimens by the separation gel tube combined with MicroflexTM matrixassisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS),perform direct antimicrobial susceptibility test based on the results of mass spectrometry,and preliminary explore the redesign of conventional positive blood culture specimen processing flow.Methods·449 positive-alarm blood culture specimens of monobacterial infections in clinical microbiology laboratory of Xirhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine from September 1,2015 to August 31,2016 were collected and prepared according to the new positive blood culture specimens processing flow.The new redesigned processing flow included smear staining and microscopy,separation gel/mass spectrometry direct identification,bacteria film/mass spectrometry identification,pure colony/mass spectrometry identification,direct antimicrobial susceptibility test,and conventional antimicrobial susceptibility test,etc.According to the results of microscopic examination,identification test,and antimicrobial susceptibility test,level Ⅰ direct microscopic examination report,positive blood culture level Ⅱ (direct identification/bacteria film identification or direct antimicrobial susceptibility) report,and level Ⅲ final report were provided sequentially.Results·With the new redesigned processing flow,bacterial specieslevel identification reports for 82.2％,95.8％,and 100％ of positive blood samples could be obtain at 10 am and 17 pm on the current day and 10 am in the next morning,respectively,and be sent to clinical departments.Initial antimicrobial susceptibility reports for the bacteria that were considered as clinical significant pathogens by preliminary assessment could be provided in the next day of blood culture positive alarm with a higher coincidence rate as compared to the results of traditional antimicrobial susceptibility tests.Conclusion·The time from blood culture positive alarm to the provision of initial identification and antimicrobial susceptibility reports is shorter by 1-2 d for the redesigned processing flow than for the traditional one,which can provide fast and accurate etiologic diagnosis evidence for bloodstream infections for clinicians and is important for improving early diagnosis and treatment of clinical bloodstream infections.

Objective To evaluate and compare yeast identification ability between Bruker Microflex matrix-assisted laser desorption ionization-time of flight mass spectrometry( MALDI-TOF MS) and Vitek 2 Compact automatic microbial analysis system.Methods Retrospective study.Totally 742 strains of yeast isolated from clinical specimens during March 2013 to March 2014 in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine were identified by Bruker Microflex MALDI-TOF MS and Vitek 2 Compact automatic microbial analysis system simultaneously.The strains with discordant results were validated by gene sequencing.Results The coincidence rate of 699 Candida identified by Bruker Microflex MALDI-TOF MS or Vitek 2 Compact system was 100.0%(699/699) and 99.6%(696/699) to the species level, respectively and the coincidence rate of 43 yeast-like fungi strains identified was 90.7%(39/43) and 79.1%(34/43) to the species level, respectively.Penicillium marneffei could not be identified by both two instruments, but protein profile of Penicillium marneffei by MALDI-TOF MS was established.Conclusions The coincidence rate of yeast identified by Bruker Microflex MALDI-TOF MS is higher than that of Vitek 2 Compact system.Using Bruker Microflex MALDI-TOF MS to identify yeast especially Candida and yeast-like fungus is fast, simple, low-cost, accurate, and it can be used in routine work of ordinary yeast identification in clinical microbiology laboratory.

AIM:To investigate the effects of celecoxib on viability , apoptosis and autophagy in acute myeloid leukemia (AML) cell lines HL-60 and HL-60A.METHODS:The HL-60 cells and HL-60A cells were cultured with vari-ous concentrations (0, 20, 40, 60, 80 and 100μmol/L) of celecoxib.The inhibitory effect of celecoxib on the cell viabil-ity was evaluated by MTT assay .Apoptosis was analyzed by Annexin-V/PI staining.Apoptosis-related and autophagy-relat-ed proteins were determined by Western blot .RESULTS:IC50 of celecoxib were 49.4 μmol/L, 32.0 μmol/L and 25.1μmol/L for HL-60 cells treated with celecoxib for 24 h, 48 h and 72 h, respectively.For HL-60A cells, the corresponding IC50 were 69.1 μmol/L, 42.5 μmol/L and 29.6 μmol/L, respectively.The results of flow cytometry analysis showed the proportions of Annexin-Ⅴ+PI-, Annexin-Ⅴ+PI+and Annexin-Ⅴ-PI+cells were increased in the HL-60 cells, and those of Annexin-Ⅴ+PI-and Annexin-Ⅴ+PI+cells were increased in the HL-60A cells treated with celecoxib for 24 h. After treated with celecoxib , the induction of apoptosis was observed , the apoptosis-related proteins cleaved caspase-3 and cleaved PARP were upregulated , the autophagy-related proteins LC3 II and P62 were both increased , and mTOR, p-mTOR, 4-EBP and p-4-EBP were not changed , indicating that celecoxib inhibited autophagy in the AML cells without the mTOR pathway involvement .CONCLUSION:Celecoxib inhibits the viability of HL-60 cells and HL-60A cells in a time-and dose-dependent manner by its effects of inducing apoptosis and necrosis .Celecoxib inhibits mTOR-independent autoph-agy in AML cells, indicating a possible way of using celecoxib for enhancing the antitumor activity of therapeutic agents to induce cytoprotective autophagy in the AML cells .

Objective To compare the efficacy of urinary type Ⅳ collagen( Ⅳ C), N-acetyl-β-D-glucosaminidase( NAG ), serum cystatin C ( CysC ), β2 microglobulin ( β2-MG ) in early diagnosis of diabetic nephropathy(DN) ,and to develop a multiple regression equation using above mentioned indices. Methods One hundred and eight cases of DM patients were enrolled in the study. All those DM patients were divided into two groups according to 24 hr urinary albumin excretion(UAE): non-DN group( UAE ＜30 mg/24 h)and DN group (UAE ≥30 mg/24 h). Receiver operating characteristics (ROC)curve was developed using urinary IVC, NAG,serum CysC and β2-MG,and the efficiency of the four indices for early diagnosis of diabetic nephropathy were assessed by area under the curve ROC (AUCROC). Furthermore, the regression equation of four indicators was developed. All statistical analyses were performed using SPSS 13.0. Results The levels of urine Ⅳ C, NAG,CysC,β2-MG,were(3.91±1.93)ng/ml, ( 12.20 ±3.46)U/L, ( 1.18 ±0.41 )mg/L , (2. 50 ±0. 74)mg/ml in the non-DN group, respectively; and ( 14.14 ± 11.17 ) ng/ml, ( 23.12 ± 13.57 ) U/L, ( 2.69 ± 1.69 ) mg/L and(5.21 ± 2.78)mg/ml in the DN group, respectively. There were significant differences in the comparison of the four indicators between the two groups ( Ps ＜ 0.01 ). AUCROC of Ⅳ C, NAG, CysC and β2-MG were 0. 747,0.732,0.764 and 0.823 respectively;which meant the diagnostic efficacy for DN decended from β2-MG, CysC,Ⅳ C, to NAG in order. All these indices showed significant efficiency in assisting diagnosis of early DN ( Ps ＜0.01 ). The regression equation of UAE and the four indices was: UAE = - 242.624 + 6.362IVC + 8.662NAG + 64. 622CysC + 29.488β2-MG, and the equation had statisticl significance( P ＜ 0.O1 ). Conclusion Urine Ⅳ C, NAG,serum CysC, and β2-MG showed significant value in assisting diagosis of early DN, and could be sensitive indices for DN.

Objective To study the clinical significance of urinary type Ⅳ collagen(IVC)and UmAlb/UCr ratio in the diagnosis of early diabetic nephropathy. Methods We collected 52 cases of diabetes(group A)without DN(UAE ＜30 mg/24 h),35 cases of diabetes(group B)with early DN(UAE as 30-300 mg/24 h),and 50 cases of healthy controls. The differences of urine IVC,UmAlb/UCr were compared among group A,B and the control group. ROC curve was used for evaluating the use of urine IVC and UmAlb/UCr in the diagnosis of early DN. The correlation of urine IVC and UmAlb/UCr with UAE were investigated,and linear model curve were established. IVC in urine was detected by chemiluminescence,UmAlb was detected by immunoturbidimetric assay,UCr was detected by enzymatic. Statistical analysis were performed with SPSS13.0 statistical software. Results The urine IVC testing of group A,B and the control group were(2. 64 ± 0. 91),(3.91 ± 1.93)and(10. 08 ± 6. 50)μg/L,respectively. The UmAlb/UCr(mg/mmol)testing of group A,B and the control group were(1.50 ± 0. 40),(2. 58 ±2. 10)and(17.95 ± 13. 38)mg/mmol,respectively. Urine IVC and UmAlb/UCr were significant difference between group A,B and the control group(Ps ＜ 0. 01);the ROC area under the curve(AUCROC)of urine IVC and UmAlb/UCr were 0. 724,0. 945,the two indicators for early diagnosis of DN were significant(Ps ＜ 0. 01);The pearson correlation coefficients of the urine IVC and UmAlb/UCr were 0. 529,0. 919 ,respectively. They were positive and significant correlation(Ps ＜ 0. 01),On the basis of the correlation coefficient and linear model fitting curve with the UAE,the relationship of UmAlb/UCr with the UAE was better than that of IVC. Conclusions Urine IVC and UmAlb/UCr ratio,which significantly assists diagnosis in diabetic nephropathy ,can be used as a sensitive diagnostic indicator of diabetic nephropathy. Moreover,the relationship of UmAlb/UCr with the UAE is better than that with IVC.

Objective To explore the risk factors of postoperative pulmonary infection in patients with craniocerebral injury and establish a nomogram model to predict the risk of postoperative pulmonary infection after craniocerebral injury.Methods The clinical data of 169 patients with craniocerebral injury,admitted to and underwent craniotomy in our hospital from January 2013 to December 2018,were retrospectively analyzed.The clinical data of patients with postoperative pulmonary infection and without postoperative pulmonary infection were compared.The risk factors of postoperative pulmonary infection were analyzed by multivariate Logistic regression.R language was used to establish a nomogram model to predict the risk of postoperative pulmonary infection after craniocerebral injury.Receiver operating characteristic (ROC) curve was used to explore the prediction efficiency of the nomogram model for pulmonary infection after craniocerebral injury.Results Among the 169 patients,74 (43.8％) were complicated with pulmonary infection and 95 (56.2％) were not complicated with pulmonary infection.As compared with non-pulmonary infection group,pulmonary infection group had significantly higher percentages of patients with open craniocerebral injury and Glasgow coma scale (GCS) scores＜7,significantly higher American Society of Anesthesiologists (ASA) grading,lower albumin level one week after surgery,statistically longer operation time,and significantly higher percentages of patients with conscious disorder,patients accepted intraoperative blood transfusion,patients used breathing machine,and patients stayed in bed for 4 weeks or more (P＜0.05).Multivariate Logistic regression analysis showed that GCS scores (OR=0.243,95％CI:0.122-0.497,P=0.000),ASA grading (OR=3.349,95％CI:2.233-5.021,P=0.000),disturbance of consciousness (OR=3.185,95％CI:1.217-8.334,P=0.018),and useofventilator (OR=3.376,95％CI:1.590-7.167,P=0.002) were independent risk factors for postoperative pulmonary infection in patients with craniocerebral injury.The scores of the nomograrn model were 13.7,100.0,38.0 and 27.5 in GCS scores,ASA grading,disturbance of consciousness and use of ventilator,respectively.The consistency index of the nomogram model for predicting postoperative pulmonary infection in patients with craniocerebral injury was 0.835.ROC curve showed that the area under the curve predicted by nomogram model for postoperative pulmonary infection in patients with craniocranial injury was 0.840 (95％CI:0.778-0.901).Conclusion Based on the risk factors for pulmonary infection after craniocerebral injury,a nomogram model for predicting the risk of pulmonary infection is established,which has a good differentiation degree and prediction effect,and can provide a reference for medical staffto identify high-risk patients at an early stage,so as to take more targeted intervention measures.

Objective:To explore the method of ultrasonography for detecting the fetal umbilical vein diameter, blood flow volume and normalized volume blood flow and establish normal reference ranges with umbilical vein diameter, blood flow volume and normalized blood flow and Z-scores for umbilical vein diameter and blood volume flow.Methods:This was a prospective study on 907 normal fetuses in the Second Xiangya Hospital, Central South University and Women and Children Healthcare Hospital of Zhuzhou from March 2019 to December 2020. The umbilical vein diameter (Duv), umbilical vein blood flow volume (Quv) and normalized volume blood flow (nQ = Quv/estimated fetal weight) of the free loop of umbilical vein (FUV) and fetal intra-abdominal umbilical vein (IUV) were collected. And the mean values and 90% confidence intervals of Duv, Quv and nQ in two segments of umbilical veins at different gestational ages were calculated. Regression analysis of Duv, Quv and nQ were performed with gestational age (GA), and the parameters of umbilical vein in different segments were compared. Finally, with gestational age (GA) as the independent variable, Z-scores of the Duv and Quv were built.Results:The mean values and 90% confidence intervals of Duv, Quv, and nQ in 858 (94.6%) normal fetal umbilical veins were successfully obtained. The Duv, Quv of the FUV and IUV increased as pregnancy progressed. The Quadratic curve of Duv and Linear curve of Quv were of the highest fitnesses, respectively( r=0.951, 0.941, 0.986, 0.982; all P<0.001). While nQ increased with GA followed by a decreased trend, and the Quadratic curve was the highest fitting curve of nQ( r=0.610, 0.611; all P<0.001). Duv-FUV was greater than Duv-IUV( P<0.001), nQ-FUV was bigger than Quv-IUV( P=0.001), and he difference was not statistically significant between Quv-FUV and Quv-IUV( P=0.133). Z-scores models of Duv and Quv were successfully established, and all Z-scores were Gaussian distribution. Conclusions:The normal ranges and Z-scores of umbilical vein parameters are useful to improve the evaluation of placental circulation and provide a strong basis for the monitoring of fetus-related diseases and the evaluation of pregnancy prognosis. The choice of FUV or IUV umbilical vein to evaluate placental circulation may depend on the actual situation in clinical application.

Objective:To explore the hematological phenotype and genotypic characteristics of five Chinese individuals with a rare thalassemia mutation HBB: c. 93-21G>A. Methods:A retrospective study was carried out on five individuals identified by the People′s Hospital of Yangjiang and Guangzhou Hybribio Co., Ltd. from May 2018 to September 2022. Routine blood test and hemoglobin electrophoresis were performed, and the genotypes of five subjects were determined by using PCR combined with reverse dot blotting (RDB), nested PCR, Gap-PCR and Sanger sequencing. This study was approved by Medical Ethics Cornmittee of the People′s Hospital of Yangjiang (Ethics No. 20240001).Results:Among the five individuals, hematological data of one was unavailable, and the remaining four had presented with microcytosis and hypochromia. The results of hemoglobin electrophoresis indicated that all of them had a HbA 2 level of ≥4.7%. Genetic analysis showed that one case had harbored compound heterozygous mutations of ααα anti3.7 triplet and HBB: c. 93-21G>A, one had compound heterozygous mutations of -α 3.7 and HBB: c. 93-21G>A, whilst the remaining three were heterozygous for the HBB: c. 93-21G>A mutation. Conclusion:The hematological phenotype of β-thalassemia carriers ( HBB: c. 93-21G>A) is similar to that of other β + thalassemia heterozygotes with mild β-thalassemia characteristics.