1.Determination of serum aromatic amino acid in patients with CRI by high performance liquid chromatography with fluorescence detection
Ying LI ; Aiguo TANG ; Sa MU ; Ledong XIAO
Chinese Journal of Laboratory Medicine 2011;34(9):769-774
ObjectiveA HPLC-FLD method was developed to determine the levels of serum AAA in CRIpatients, and to studythe variationof serum AAAinCRI patientsanditsclinical significances. MethodsSerumsampleswerecollected from100healthcontrolsand 80CRI patients. According to 2002 National Kidney Foundation (NKF) staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CRI patients were also divided into 3 groups :chronic nephritis group ( n = 32), DM group ( n = 36), hypertension group ( n = 12 ).Serums were deproteinized by equal volume of 5% (v/v) PCA and supernate were analyzed direcdy. External standard method was used as quantitative method. The analytical column was Megres C18. 10% acetonitrile in water was used as mobile phase. Flow rote was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. ResultsThe linear ranges of the method were 0. 550 -275.000, 3. 050 - 1220. 000 and 0. 049 -49. 000 pμmol/L for Tyr, Phe and Trp, respectively. The limit of detection (LOD) was 0.014 μmol/L for Tyr, 0.500μmol/L for Phe, and 0.005 μmol/L for Trp. The average recovery was 100. 9%, 101.3% and 98. 5% for Tyr, Phe and Trp, respectively. Intra-day CVwas 3. 18% -4. 20% ( mean was 3. 13% )and inter-day CV was 3. 18% -4. 20% ( mean was 3. 58% ). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were( 135.74 ±23.23 )μmol/L, (52.27 +8.25) μmol/L, (21.49 ±4.25) μmol/L and[0.87(0.68 - 1.05)]μmol/L. which were lower than that in healthy groups (t value was -14. 709, 4.452, 22. 100, U value was 266.000,respectively, P<0. 05). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in healthy groups were ( 174. 47 ± 11.57 ) μmol/L, ( 63.53 ± 4. 68 ) μmol/L, (44. 22 ± 3. 67 ) μmol/L and[0. 97(0. 94 - 1.00)]μmoL/L. There were no statistically significant difference between the different stage of CRI. Compared with the concentration of Tyr, Phe and Trp among chronic nephritis group, DM group,hypertension group, the concentration of Tyr had no significant changes among these three kinds of diseases (P > 0. 05 ). The concentration of Phe had significant changes between Chronic nephritis group and DM group, Chronic nephritis group and hypertension group ( U = 395.00, 114. 00, P < 0. 05 ) ; the concentration of Trp haad significant changes between Chronic nephritis group and DM group ( U = 349.00, P < 0. 05 ).The diagnostic sensitivity and specificity of serum AAA for CRI were 90% (72/80) and 100. 0% (100/100).ConclusionsThe method of high-performance liquid chromatography with fluorescence detection ( HPLC-FLD) is simple, rapid, sensitive and specific. Simultaneous determination of serum AAA was benefit to the diagnosis and evaluation of CRI patients.
2.Case-control study of risk factors for recurrent low hemoglobin level among primary and middle school students
Dan-dan HE ; Xiao-sa WEN ; Hong-mei TANG ; Di-di CHEN ; Xiao-li XU ; Hua CHENG
Shanghai Journal of Preventive Medicine 2020;32(8):675-
Objective To explore the risk factors of recurrent low hemoglobin(RLH) level among students from 6 to 13 years old, and to formulate strategies and policies in this regard. Methods Surveillance on hemoglobin concentration was conducted among 71 742 students aged from 6 to 13y between 2013-2014 based on the annual physical examination for primary and middle school in Minhang District.Of those students, 670 were diagnosed with low hemoglobin level according to WHO criteria.A 1 ︰ 1 matched case-control study was conducted based on gender, age and school type.Questionnaire surveys for data collection were analyzed using Cox′s proportional hazards regression. Results Factors as pregnancy anemia(
3.Effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network
Zhiming TANG ; Mengqing JING ; Lu LU ; Xiao SHAN ; Cuixia ZHANG ; Xiaoyu ZHANG ; Sa MENG
Chinese Journal of Dermatology 2023;56(7):642-650
Objective:To investigate the effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) /microRNA (miR) -485-5p/signal transducer and activator of transcription 3 (STAT3) regulatory network. Methods:HaCaT cells were induced by interleukin-17 (IL-17), and the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3 was detected in IL-17-induced HaCaT cells and normal human epidermal keratinocytes (NHEK) by quantitative PCR (qPCR) and Western blot analysis, respectively. The location of lncRNA NEAT1 and miR-485-5p in IL-17-induced HaCaT cells was observed by fluorescence in situ hybridization (FISH), and the targeted regulatory relationship among lncRNA NEAT1, miR-485-5p and STAT3 was verified by double-luciferase reporter gene assay. Chinese herbs were decocted according to the Xidi Liangxue recipe, SD rats were divided into two groups to be gavaged with the above decoctions (medicated group) or physiological saline (control group) for 5 days, and then serum samples were collected from the above two groups of rats separately. The IL-17-induced HaCaT cells were divided into 4 groups: control group treated with the control sera, lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the control sera, Xidi Liangxue recipe group treated with the medicated sera, and Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the medicated sera. qPCR, Western blot analysis, flow cytometry, and cell counting kit (CCK8) assay were performed to determine the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3, and to evaluate cell proliferation and apoptosis. The two independent samples t-test was used for comparisons between two groups, one-way analysis of variance for comparisons among multiple groups, and least significant difference (LSD) t-test for multiple comparisons. Results:The IL-17-induced HaCaT cell group showed significantly increased relative expression levels of lncRNA NEAT1 and STAT3 mRNA (1.84 ± 0.21, 2.20 ± 0.24, respectively) and significantly increased protein expression of STAT3 and p-STAT3 (1.27 ± 0.13, 2.43 ± 0.16, respectively), but significantly decreased expression level of miR-485-5p (0.32 ± 0.04) compared with the NHEK group (lncRNA NEAT1 and STAT3 mRNA: 1.00 ± 0.11, 1.00 ± 0.11, respectively, both P < 0.05; STAT3 and p-STAT3 protein: 1.00 ± 0.11, 1.00 ± 0.10, t = 2.54, 3.02, respectively, both P < 0.05; miR-485-5p: 1.00 ± 0.12, t = 2.94, P = 0.015). FISH demonstrated that miR-485-5p and lncRNA NEAT1 were co-located in the cytoplasm of HaCaT cells. The double-luciferase reporter gene assay showed that the relative activity of luciferase was significantly lower in the miR-485-5p group than in the negative control group (both P < 0.05) after the transfection with wild-type lncRNA NEAT1 or STAT3 recombinant plasmids, while there were no significant differences between the miR-485-5p group and negative control group after the transfection with mutant lncRNA NEAT1 or STAT3 recombinant plasmids (both P > 0.05). Compared with the control group, the lncRNA-NEAT1 overexpression group showed significantly increased expression of lncRNA NEAT1 and STAT3 (including STAT3 mRNA, STAT3 protein, and p-STAT3 protein) in HaCaT cells (all P < 0.05), but significantly decreased miR-485-5p expression ( P < 0.05) ; the Xidi Liangxue recipe group showed significantly decreased expression of lncRNA NEAT1 and STAT3 (all P < 0.05), but significantly increased miR-485-5p expression compared with the control group ( P < 0.05) ; significantly decreased expression of lncRNA NEAT1 and STAT3, but significantly increased miR-485-5p expression was observed in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group compared with the lncRNA-NEAT1 overexpression group (all P < 0.05). After 24-, 48-, and 72-hour intervention, CCK8 assay showed that the proliferative activity of HaCaT cells was significantly higher in the lncRNA-NEAT1 overexpression group than in the control group (all P < 0.05), as well as in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group than in the Xidi Liangxue recipe group (all P < 0.05), and the cellular proliferative activity was significantly lower in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group and Xidi Liangxue recipe group than in the control group (all P < 0.05). The apoptosis rate was significantly lower in the lncRNA-NEAT1 overexpression group (5.84% ± 0.28%) than in the control group (14.75% ± 0.83%, LSD- t = 3.48, P = 0.002), but significantly higher in the Xidi Liangxue recipe group (35.72% ± 3.62%) than in the control group (LSD- t = 5.34, P = 0.001) ; the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group showed significantly increased apoptosis rate (27.64% ± 2.82%) compared with the lncRNA-NEAT1 overexpression group (LSD- t = 9.06, P < 0.001) . Conclusion:The Xidi Liangxue recipe could inhibit the proliferation of IL-17-induced HaCaT cells and promote their apoptosis, which may be related to the intervention in the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network.
4.Prevalence and incidence of anemia among school students aged 6-11 years old in Minhang District of Shanghai
dan Dan HE ; sa Xiao WEN ; yi Xin ZHANG ; di Di CHEN ; mei Hong TANG ; li Xiao XU ; Hong FANG ; jing Hui SHI
Fudan University Journal of Medical Sciences 2017;44(5):633-638
Objective To analyze the epidemic features of anemia among students aged 6-11 years old in Minghang District,to provide basis for the control and prevention strategies of anemia among school students.Methods Surveillance on hemoglobin concentration was conducted among 42 872 students aged from 6 to 11 years old between 2012 and 2015.All data analyses were completed by SPSS 18.0 statistical software.Results The prevalence rate of anemia among school students 6-11 years old was 5.05% (95%CI:4.84-5.26).The prevalence rate of anemia among girls was 5.28%,which was significantly higher than that 4.84% among boys (x2 =4.24,P =0.037).Six-year-old boys and girls were most susceptible to anemia.Compared to students in Hope Schools,public school students are more vulnerable to anemia (boys:OR =2.37,95 % CI:2.03-2.76;girls:OR =2.08,95 % CI:1.74 -2.49).Overweight and obese students had a lower risk of anemia than average students (boys:OR =0.65,95 % CI:0.55-0.76;girls:OR =0.75,95 % CI:0.61-0.92).The three-year cumulative incidence of anemia was 12.80% (95 %CI:12.49-13.12).The cumulative incidence among girls was 14.52%,which was significantly higher than that of 11.28% among boys (x2 =100.26,P<0.001).Six-year-old boys have the highest three-year cumulative risk among all students,while 10-11-year-old girls have the highest three-year cumulative incidence.The risk of anemia in Hope School was found highest in all schools (boys:RR =1.93,95%CI:1.72-2.16;girls:RR =1.20,95 %CI:1.04-1.39).Overweight and obesity were considered protective factors to anemia (boys:RR =0.75,95%CI:0.67-0.84;girls:RR =0.77,95%CI:0.68-0.88).The primary and recurrent detection rates of anemia were 14.58% and 2.54%.The anemia among students was mainly detected during the first examination despite the differences in ages,schools and nutrition conditions (P < 0.05).Conclusions The prevalence of anemia among primary school students in Minhang District is at a mild level.Control measures should be adopted,including monitoring of hemoglobin in all school children and adolescents,taking individual intervention measures based on results,promoting the health education for adolescent girls and parents of school children,and paying special attention to Hope School and students with recurrent and continuous anemia.
5.A cross-sectional survey of the relationship between eating habits and childhood obesity:based on a cohort study of school children in Minhang District of Shanghai
di Di CHEN ; dan Dan HE ; sa Xiao WEN ; yi Xin ZHANG ; mei Hong TANG ; li Xiao XU ; Hong FANG ; wei Chao FU
Fudan University Journal of Medical Sciences 2017;44(5):626-632,643
Objective To investigate the relationship between eating habits and childhood obesity by the method of cross-sectional survey in order to provide scientific basis for the prevention and intervention of childhood obesity.Methods We enrolled first-year students in Minhang District of Shanghai who participated in physical examination in 2014.Information was obtained from standardized face to face interviews of their parents or guardians with detailed questions on demographic characteristics,dietary habit,food preference,physical activity,sleep time and so on.Odds ratio (OR) and 95% CI were calculated by unconditional Logistic regression models.Results Finally,5 752 students were enrolled in the study.The rate of obesity was 15.16% (boys:18.35%;girls:11.83%) and the rate of over weight was 14.31% (boys.:15.15%;girls:13.43%).Meat-based diet (OR=1.58,95% CI:1.32-1.91),skipping breakfast (OR =1.87,95% CI:1.01-3.46),eating too much caffeine-containing food (OR =1.66,95% CI:1.09-2.51) and eating fast (OR:3.32-14.95) were associated with an increased risk of childhood obesity.Plant-based diet (OR =0.51,95% CI:0.35-0.74) and choosy in food (OR:0.38-0.64) were associated with a decreased risk of childhood obesity.Besides,we found that children who preferred meat (OR =2.11,95%CI:1.42-3.14) and cured food (OR =1.42,95% CI:1.05-1.92) had a higher risk of obesity.Conclusions Our results support the association between eating habits and childhood obesity.
6.Establishment of immune suppression-mediated infectious pneumonia model using resistant Acinetobacter baumannii strains
Zining TANG ; Xuewu LIU ; Qianqing LAN ; Hailiang TANG ; Sa XIAO ; Xiangzhong YUAN ; Dongdong PENG
Chinese Journal of Microbiology and Immunology 2023;43(12):908-917
Objective:To establish an animal model of pneumonia for research on clinical prevention and treatment of bacterial pneumonia by infecting immunocompromised rats with drug-resistant Acinetobacter baumannii ( Ab) strains. Methods:Drug-resistant Ab strains were selected. Forty-eight SD rats were randomly divided into four groups: normal group, cyclophosphamide control group (intraperitoneal injection of 45 mg/kg cyclophosphamide), bacterial infection group (intratracheal instillation of 1.5×10 8 CFU Ab suspension), and bacterial infection+ immunosuppression group (intraperitoneal injection of 45 mg/kg cyclophosphamide+ intratracheal instillation of 1.5×10 8 CFU Ab suspension). Flow cytometry analysis was used to detect the proportion of CD4 + , CD8 + and NK cells in rat peripheral blood before as well as 3 d and 7 d after infection. A lung function meter was used to detect peak inspiratory flow (PIF), peak expiratory flow (PEF), tidal volume (Vt ) and forced expiratory volume in the second second/forced vital capacity (FEV 200/FVC) at 3 d and 7 d after modeling. ELISA was used to detect the levels of IL-6, TNF-α and IL-10 in the alveolar lavage fluid. HE staining was used to observe the morphology of rat lung tissues in each group. Bacterial loads in rat lung tissues were counted by bacterial culturing. Results:A decrease in voluntary activity was observed in rats in the cyclophosphamide control group, bacterial infection group and bacterial infection+ immunosuppression group after modeling. Lung rales could be heard in the bacterial infection group and bacterial infection+ immunosuppression group. Compared with the normal group, the cyclophosphamide control group showed decreased proportion of CD4 + and CD11b + NK cells and increased CD8 + cells in peripheral blood; the bacterial infection group showed decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in the alveolar lavage fluid, and higher bacterial load in lung tissues with mild widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). Compared with the cyclophosphamide control group and the bacterial infection group, the bacterial infection+ immunosuppression group showed a lower proportion of CD4 + cells and a higher proportion of CD8 + cells in rat peripheral blood, decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in alveolar lavage fluid, higher bacterial load in lung tissues, and aggravated widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). The proportion of CD11b + NK cells in peripheral blood of rats in the bacterial infection+ immunosuppression group was significantly lower than that in the bacterial infection group ( P<0.05, P<0.01). Conclusions:A bacterial pneumonia model was successfully constructed by infecting rats with Ab alone or in combination with cyclophosphamide immunosuppression. In the model constructed with Ab and cyclophosphamide immunosuppression, the rats had more severe pneumonia, which might be related to the reduced cellular immune function and the aggravated bacterial infection in rat lung tissues by cyclophosphamide.
7.Research of malnutrition epidemiologic trend among students aged 6-18 years from 2010 to 2016 in Minhang District of Shanghai
yi Xin ZHANG ; dan Dan HE ; Hong FANG ; sa Xiao WEN ; di Di CHEN ; Qi GUO ; mei Hong TANG ; li Xiao XU ; Na WANG
Fudan University Journal of Medical Sciences 2017;44(5):621-625
Objective To analyze malnutrition epidemiologic trend among students aged 6-18 years from 2010 to 2016 in Minhang district of Shanghai,and to provide reference for malnutrition prevention and treatment.Methods Physical examination data from healthy monitoring schools in Minhang District from 2010 to 2016 were used to screened out the malnutrition of students aged 6-18 years via National standard 2014 and analyzed the different type of malnutrition in different population.Results During the period of 2010 to 2016,malnutrition rate of students demonstrated a fluctuated trend.Malnutrition rate in 2010 was 4.0% and 4.4% in 2016.The major type of malnutrition is wasting with a proportion more than 88% during these years.Rate of wasting increased from 3.5% to 7.8% in boys aged 13-18 years old and from 1.3% to 3.9% in girls aged 13-18 years old.Conclusions The malnutrition of students in Minhang District is under control,while the screening rate of wasting increased in students aged 13-18 years old.Intensified intervention of prevention and treatment should be administrated and integrated with obesity control in Minhang District of Shanghai.
8.Risk factors of persistent thrombocytopenia after adult liver transplantation and prophylactic measures.
Long-Yu CHENG ; ; Jun YU ; ; Wu ZHANG ; ; Song-Feng XIE ; ; Lin ZHOU ; ; Min ZHANG ; ; Sa-Xiao TANG ; ; Lin ZHANG ; ; Shu-Sen ZHENG ;
Journal of Zhejiang University. Medical sciences 2014;43(6):670-677
OBJECTIVETo investigate the risk factors associated with persistent thrombocytopenia after liver transplantation (LT), and to explore effective measures for prevention.
METHODSOne hundred and twenty-eight adult patients, who received liver transplantation in our hospital between January 2009 and June 2012 and met the inclusive criteria, were enrolled in the study. The clinical data were retrospectively analyzed, including pre-LT spleen volume, main portal vein size, coronary vein size, platelet and white blood cell levels, total bilirubin level and model of end stage liver disease score. The risk factors associated with persistent thrombocytopenia after LT were evaluated by logistic regression analysis. The effect of simultaneous splenic artery coarctation for high risk patients was evaluated with χ2 test.
RESULTSLogistic regression analysis showed that per-LT spleen volume larger than 500 ml (P = 0.012, OR=2.789, 95%CI: 1.249-6.227) and portal vein size beyond 15 mm (P = 0.017, OR = 3.124, 95%CI: 1.230-7.933) were independent risk factors for persistent thrombocytopenia after LT. The incidence rate of persistent thrombocytopenia after LT in patients with or without simultaneous splenic artery coarctation were 16.7% (1/6) and 66.7% (32/48), respectively(P < 0.05).
CONCLUSIONSpleen volume larger than 500 ml and portal vein size beyond 15 mm are risk factors for persistent thrombocytopenia after LT. Simultaneous splenic artery coarctation may reduce the occurrence of persistent thrombocytopenia after LT.
Adult ; Humans ; Liver Transplantation ; adverse effects ; Organ Size ; Portal Vein ; anatomy & histology ; Retrospective Studies ; Risk Factors ; Spleen ; anatomy & histology ; Thrombocytopenia ; epidemiology