OBJECTIVE To establish and validate a rat liver micronucleus test(LMNT)method based on fixation of liver tissue with 4%neutral formaldehyde(HCHO fixation)for preparation of hepa-tocytes(HEPs).METHODS ①The LMNT based on neutral HCHO fixation(HCHO fixation-LMNT)was established using the liver micronucleus positive compound N-nitrosodiethylamine(DEN).SD rats were divided into female and male groups,and each group was randomly subdivided into the vehicle control group and DEN 12.5 mg·kg-1 group,with five rats in each.The rats were ig administered with normal saline and DEN once a day for 14 consecutive days,after which liver tissues were collected.Some of the tissue was digested with collagenase to prepare HEP suspension,and the remaining tissue was used to prepare HEP suspension with HCHO fixation.After staining with SYBR Gold,the number of micronucleated hepatocytes(MN-HEP)and the number of HEPs in the mitotic phase were counted under a microscope.The micronucleus rate of HEP(MN-HEP rate)and the mitotic index were calculated,and an MN-HEP rate>0.07%was considered positive.②Male SD rats were divided into the quinoline(30,60,120 mg·kg-1)group,N-nitrosoopyrrolidine(NPYR,25,50,100 mg·kg-1)group,vehicle control group(deionized water for NPYR,and corn oil for quinoline),and positive control DEN(12.5 mg·kg-1)group,with 5-6 rats per group,and were ig administrated for 15 consecutive days.Body mass was recorded daily,and at the end of the experiment,the liver was removed to record the total liver weight,and calculate the liver coefficient.Liver function-related serum biochemical indicators including glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT)activities,and levels of total bili-rubin(T-BIL)were measured and direct bilirubin(D-BIL)using an automatic biochemical analyzer.The MN-HEP rate was determined using the collagenase digestion and HCHO fixation methods,and the peripheral blood MN assay and hepatocellular carcinoma comet assay were conducted to evaluate the genotoxicity of quinoline and NPYR.RESULTS ① Compared with the corresponding vehicle control groups(0.069%and 0.030%),the MN-HEP rate of male rats treated with DEN by formalin-LMNT was 1.10%,and the MN-HEP rate of female ones was 0.82%,both significantly increased(P<0.05).Com-pared with corresponding vehicle control groups(0.060%and 0.030%),the MN-HEP rate of male rats treated with DEN by collagenase digestion-LMNT was 1.45%,and that of female rats was 0.46%,both significantly increased(P<0.05),which were considered positive.The MN-HEP rate of male rats was significantly higher than that of females with both methods(P<0.05).There was no significant differ-ence in mitotic indexes between the DEN groups by collagenase digestion-LMNT and HCHO fixation-LMNT in male and female rats compared to corresponding vehicle control groups.② Compared to the vehicle control group,the body mass of rats in the NPYR 50 and 100 mg·kg-1 groups was significantly reduced 7 to 14 days into the ig administration(P<0.01),and the DEN group showed a significant reduction at days 8 to 14(P<0.01).The body mass of rats in the quinoline 120 mg·kg-1 group was signifi-cantly reduced 4 to 14 days into the ig administration(P<0.01),and the DEN group showed a signifi-cant reduction at days 10 to 14(P<0.05).Compared to the vehicle control group,both the liver weight and liver coefficient were significantly reduced in the NPYR 100 mg·kg-1 group(P<0.01)and the DEN group(P<0.05).The liver weight(P<0.01)and liver coefficient(P<0.05)were significantly increased in the quinoline 60 and 120 mg·kg-1 groups.Compared to the vehicle control group,the serum T-BIL level was significantly increased in the DEN group(P<0.01),and the activities of GPT and GOT,as well as the levels of D-BIL and T-BIL,were significantly increased in the NPYR 100 mg·kg-1 group(P<0.01).There were no significant changes in the NPYR 25,50 mg·kg-1 groups or any of the dose groups of quinoline.The MN-HEP rate by HCHO fixation-LMNT for NPYR was slightly higher than that by collage-nase digestion-LMNT,both considered positive.Compared with corresponding control group,the MN-HEP rate by formalin-LMNT for NPYR and the MN-HEP rate by collagenase digestion for NPYR were both significantly increased(P<0.05).The MN-HEP rate by HCHO fixation-LMNT for quinoline was comparable to that by collagenase digestion-LMNT,both considered positive.Compared with corresponding vehicle control group,the MN-HEP rate by HCHO fixation-LMNT for quinoline and the MN-HEP rate by collagenase digestion-LMNT for quinoline were both significantly increased(P<0.05).The correlation between the MN-HEP rates based on HCHO fixation and collagenase digestion for NPYR and quinoline was good(R2=0.8614 and 0.9279,respectively).In the NPYR groups,the periph-eral blood micronucleus assay were negative,while the comet assay results were positive.In the quino-line group,both the peripheral blood micronucleus assay and the comet assay results were negative.CON-CLUSION The HCHO fixation-LMNT has been established and validated,and the sensitivity of the LMNT method based on HCHO fixation-LMNT for detection of hepatocarcinogens is higher than that of collagenase digestion-LMNT.

Objective:To evaluate the left ventricular myocardial strain in patients with light chain cardiac amyloidosis (AL-CA) with normal left ventricular ejection fraction (LVEF) by three-dimensional speckle tracking imaging(3D-STI), and to explore the clinical value of 3D-STI in predicting the prognosis of AL-CA patients with normal LVEF.Methods:A total of 80 patients with AL-CA and LVEF≥50% were retrospectively analyzed in the Xijing Hospital of Air Force Military Medical University from October 2014 to May 2020.According to whether the patients had endpoint events, they were divided into endpoint event group and non-endpoint event group. The clinical data, conventional echocardiographic parameters, 3D-STI related parameters and follow-up results were collected. Cox regression proportional hazards model was used to analyze the survival status of AL-CA patients with univariate and multivariate regression analyses, in order to find the relevant indicators of conventional echocardiography and 3D-STI to predict adverse events.Results:All patients were followed up for 20(7.3, 40.8) months. At the end of follow-up, 25 patients had all-cause deaths. Compared with the non-endpoint group, the endpoint event group had significantly increased left ventricular end diastolic maximum wall thickness (MLVWT), peak early diastolic flow velocity/peak early diastolic velocity at mitral annulus(E/e′) (all P<0.05), and decreased LVEF, left ventricular global longitudinal strain (GLS) and basal segment longitudinal strain (LS) (all P<0.05). Multivariate cox regression analysis after adjusting for age and gender showed that basal segment LS ( HR=0.812, 95% CI=0.675-0.976, P=0.026) was an independent predictor of end-point events in patients with AL-CA. Kaplan-Meier survival curve showed that AL-CA patients with basal segment LS≤13.07% were more likely to have endpoint events. Conclusions:Basal segment LS can be used as a predictor of endpoint events in patients with AL-CA.

Objective:To explore the value of three-dimensional speckle tracking imaging (3D-STI) in the diagnosis of immunoglobulin light-chain cardiac amyloidosis(AL-CA) patients with normal left ventricular ejection fraction (LVEF).Methods:A total of 92 consecutive patients diagnosed with systemic immunoglobulin light chain amyloidosis(sAL) and with normal LVEF from October 2014 to January 2018 in Xijing Hospital were enrolled.Based on the diagnostic criteria of cardiac involvement, the patients were divided into AL-CA group (52 cases) and immunoglobulin light chain amyloidosis (AL) group (40 cases). The clinical data and serological markers of the patients were collected, the conventional echocardiography and full-volume three dimensional dynamic images were acquired, left ventricular global longitudinal strain (GLS), global radial strain (GRS), global circumferential strain (GCS), and global area strain (GAS) were analyzed using off-line TomTec software. The differences between the two groups were compared.Results:Compared with the AL group, the NT-proBNP of AL-CA group was significantly higher ( P<0.05) and there were no significant differences of the other serological indexes between the two groups(all P>0.05). Compared with the AL group, the maximal left ventricular wall thickness, left ventricular mass index, left atrial volume index, and E/e′ in the AL-CA group were significantly increased (all P<0.05). There were no significant differences of other conventional echocardiographic measurements between the two groups(all P>0.05). Compared with the AL group, GLS, GAS, and GRS were significantly lower in AL-CA group (all P<0.05); but there was no significant difference of GCS between the two groups( P>0.05). The ROC curve analysis showed that the cut-off values discriminating cardiac involvement were 16.09% for GLS, 36.54% for GAS and 31.90% for GRS. Conclusions:3D-STI measurements of left ventricular myocardial mechanics could detect cardiac involvement in patients with sAL amyloidosis, and provides a new method for diagnosis of AL-CA.

Objective: Previous cross-sectional studies suggested that elevated levels of total cholesterol content of erythrocyte membrane (CEM) could significantly increase the risk of acute coronary syndrome (ACS). The purpose of the present study was to assess the predictive value of baseline CEM levels for the risk of clinical endpoint events in patients with ACS through prospective follow-up studies. Methods: This study is a prospective follow-up study, which consisted of 859 patients with first ACS (698 patients with unstable angina pectoris and 161 patients with acute myocardial infarction), diagnosed and hospitalized in the First and Second Affiliated Hospital of Anhui Medical University. The routine blood lipid levels and CEM were measured. Patients were divided into two groups according to the median of baseline CEM: CEM≤131.56 μg/mg group (n=430) and CEM>131.56 μg/mg group (n=429). Patients were followed up at 6 months interval. The clinical endpoints were nonfatal myocardial infarction, nonfatal stroke, all-cause mortality, all-cause mortality, heart failure requiring hospitalization, and coronary artery revascularization. Kaplan-Meier curve analysis and Cox proportional hazard model were used to analyze the impact of elevated CEM on the occurrence of clinical end-point events. HR values and 95%CI of each variable were obtained. Cox regression analysis of all-cause mortality was performed according to whether patients had risk factors for coronary heart disease (hypertension, diabetes, smoking and elevated LDL-C) and whether they were treated with PCI. Results: The follow-up time was 1 640 (1 380, 2 189) days. Cox analysis after adjustment showed that an elevated baseline of CEM (>131.56 μg/mg) was associated with an increased risk of all-cause mortality (HR=1.690, 95%CI 1.041-2.742, P=0.034), but had no significant predictive effect on the other clinical endpoints. Subgroup analysis showed that elevated baseline CEM levels in ACS patients with LDL-C>1.8 mmol/L (HR=1.687, 95%CI 1.026-2.774, P=0.039), receiving in-hospital PCI (HR=2.365, 95%CI 1.054-5.307, P=0.037), or male (HR=1.794, 95%CI 1.010-3.186, P=0.046) were associated with an increased risk of all-cause mortality. Conclusion The results showed that elevated CEM levels can increase the risk of all-cause mortality in ACS patients.

Objective To investigate the effect of kinesio taping on upper crossed syndrome (UCS). Methods From December, 2016 to March, 2017, 52 patients with UCS were randomly divided into control group (n=26) and observation group (n=26). The control group re-ceived exercise therapy, and the observation group received kinesio taping in addition, for four weeks. They were assessed with Visual Ana-logue Scale (VAS), forward head angle (FHA) and forward shoulder angle (FSA). Results There was no significant difference in the score of VAS, FHA and FSA between two groups before treatment (P>0.05). After treatment, the scores of VAS, FHA and FSA significantly de-creased (t>2.804, P<0.05), and they were less in the observation group than in the control group (t>2.632, P<0.05). Conclusion Exercise therapy could relieve pain and correct abnormal posture of UCS, which is more effective combined with kinesio taping.

OBJECTIVE To investigate the delayed cytotoxicity effect of chlorpyrifos (CPF) with?drawal on primary hippocampal neurons. METHODS Hippocampal neurons were prepared from SD rat fetuses on the 17th day of gestation. Seven days after culture,neurons were exposed to CPF 10 and 30 μmol · L-1,respectively,for 72 h or for 48 h followed by CPF withdrawal for 24 h. CCK-8 kit and neuronal nuclei(NeuN), 5-bromodeoxyuridine(BrdU) and β Ⅲ tubulin immunofluorescence expression methods were used to evaluate the cell viability. RESULTS Compared with normal control, no significant cytotoxicity was found after CPF 72 h continuous exposure. However,CPF 48 h expo?sure followed by CPF withdrawal for 24 h induced evident cytotoxicity. The amount of BrdU positive and β Ⅲ tubulin positive hippocampal neurons were both decreased significantly(P<0.05),and cell survival and viability reduced after CPF withdrawal. CONCLUSION CPF exposure withdrawal can induce more seriously delayed cytotoxicity than continuous exposure in rat primary hippocampal neurons.

OBJECTIVE: To explore the prevalence of anemia during pregnancy and the influential factors in changsha city, and to provide scientific evidence for prevention and intervention of anemia during pregnancy. 
 METHODS: A cluster sampling of 713 mothers, who delivered baby between January 1, 2013 and December 31, 2013 in the three streets of kaifu district of Changsha, was selected to establish a retrospective cohort. The maternal health information during pregnancy was collected within 15 days after the delivery. The influential factors of anemia during pregnancy were analyzed by the method of retrospective nested case-control study and conditional logistic regression.
 RESULTS: The prevalence of anemia during pregnancy in Changsha was 9.96%. The multiple conditional logistic regression results show that high family income (OR=0.632, P=0.020), high education level (OR=0.276, P=0.033), folic acid supplement (OR =0.248, P=0.050), iron supplement (OR=0.272, P<0.001) and eating blood products (OR=0.588, P=0.044) were the beneficial factors for anemia during pregnancy, while multipara (OR=2.917, P=0.003), long duration of menstrual period before pregnancy (OR=1.335, P=0.041), living in new decoration housing (OR=3.690, P=0.045) and tea-drinking (OR=1.365, P=0.094) were the risk factors for anemia during pregnancy. 
 CONCLUSION The prevalence of anemia during pregnancy in Changsha is below the average levels. The occurrence of anemia during pregnancy is closely related to the women's economic level and nutritional status. Women should strengthen maternal nutrition during pregnancy and bedroom environment monitor, drink little strong tea and improve the nutritional status of iron during pregnancy.
Anemia ; Case-Control Studies ; Dietary Supplements ; Female ; Humans ; Iron ; Nutritional Status ; Pregnancy ; Prevalence ; Retrospective Studies ; Risk Factors

OBJECTIVE: To compile a questionnaire on maternal and child health birth cohort study, to provide effective tools for exploring the effect of various exposure factors on pregnancy outcome and children's health, and to track children's growth and development dynamically.
 METHODS: The preliminary questionnaire on maternal and child health birth cohort study was designed after literature research, health information systems analysis and group discussion. The questionnaire was finalized after expert consultation and pre-survey.
 RESULTS: The formal questionnaire on maternal and child health birth cohort study was developed, including maternal health information and children's health information.
 CONCLUSION The questionnaire can be used for collecting maternal health data and children's health data within 1 year old, and this work is useful for scholars in the study on birth cohort invovling the maternal and child health.
Child Health ; Cohort Studies ; Female ; Humans ; Infant ; Maternal Health ; Pregnancy ; Pregnancy Outcome ; Surveys and Questionnaires

OBJECTIVE: To explore the impact of pre-pregnancy body mass index on baby's physical growth and nutritional status. METHODS: A total of 491 pairs of mother-infant were divided into 3 groups according to mother's pre-pregnancy body mass index (BMI): a pre-pregnancy low BMI group (BMI<18.5 kg/m², n=93), a pre-pregnancy normal BMI group (18.5 kg/m² ≤ BMI<24.0 kg/m², n=326), and a pre-pregnancy high BMI group (BMI ≥ 24.0 kg/m², n=72). Analysis of variance of repeated measurement data and the median percentage methods were used to compare the physical growth and nutritional status of babies in different groups. RESULTS: Baby's weight in the high BMI group were higher than that in the normal BMI and the low BMI group (F=3.958, P=0.020). The incidence of malnutrition in the low BMI group showed a tendency to decline along with the months (χ²=5.611, P=0.018), the incidence of overweight and obesity in the high and the normal BMI groups displayed a tendency to decline along with the months (χ²=18.773, 53.248, all P<0.001). Baby in the low BMI group had higher incidence of malnutrition while baby in the high BMI group had higher incidence of overweight and obesity. CONCLUSION Pregnancy BMI was correlated with the growth of baby. Too high or too low prepregnancy BMI exerts harmful effect on baby's weight and nutritional status. Medical workers should strengthen the education on women's pre-pregnancy to remind them keeping BMI at normal level.
Birth Weight ; Body Mass Index ; Female ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Nutritional Status ; Obesity ; Overweight ; Pregnancy ; Weight Gain

Objective To investigate the status of treatment institution selection and its influencing factors in elderly patients with chronic diseases in a community in Changsha.Methods By using systematic sampling,400 elderly patients with chronic diseases from June to August 2012 were included in the investigation by questionnaires.The influencing factors were analyzed by binary logistic regression model or multinomial logistic regression model on the basis of the numbers of response variables.Results In mild conditions in self-consciousness,more than half of the elderly patients with chronic diseases (206 cases,51.5％) preferred community health service centers including private clinics (67 cases,16.8％),municipal hospitals (92 cases,23.0％),third-grade class-A hospitals (35 cases,8.8％).In serious conditions in self-consciousness,patients preferred third-grade class-A hospital (238 cases,59.5％) and municipal hospitals (162 cases,40.5％),no one selected the private clinics or community health service center (x2=443.240,P＜0.001).The analysis of impact factors for the first treatment institution selection in patients with serious conditions in self-consciousness showed that,compared with patients selecting the municipal hospital,the following personnel preferred to choose third-grade class-A hospital:patients in the staff remaining in positions (OR=32.38) and retirees (OR=15.17),patients with monthly income more than 5,000 yuan (OR=134.28),rural cooperative medical patients (OR=661.42),patients with highly trusting in community health service centers (OR=4.23),patients with BP and serum glucose determination at a frequency of once a month to every three months (OR=83.93),patients unknowing the telephone numbers of community health service centers(OR=3.02)and patients living far away from community health service centers (OR=15.70),patients with little information about community health service centers (OR=1.62).The analysis of impact factors for the first treatment institution selection in patients with mild conditions in self-consciousness showed that,compared with patients selecting the private hospitals,the following personnel preferred to choose community health service centers:patients with primary education level (OR=0.15),retired persons (OR=0.14),patients with higher health attention (OR=0.26),while persons with less health attention (OR-10.65) and patients living far away from community health service center (OR =5.22) were more willing to choose the private hospitals.Conclusions Patient selection for treatment agencies is affected mainly by the perceived severity,in addition,age,education level,employment status,income,health insurance type,trust and understanding on community health services centers,and the emphasis on health and the distance from community health services centers.