Objective: To find out the relation factors of hyperuricemia in cadres. Methods: The cadres who had health check on 2008 were analyzed. Results: The rate hyperuricemia in cadres of Zhuhai city was 35.42%. Conclusion: The rate of hyperuricemia in cadres is increasing by ages. It has the relations with sexual distinction, overweight or obesity, abnormal triglyceride and high purine-food.

BACKGROUND:FGF2,TGFβ/activin/nodal and IGF signal pathways are necessary for keeping functions of human embryonic stem calls,but there was no report concerning whether addition of exogenous basic fibroblast growth factor,transforming growth factor β and insulin can maintain self-renewal of human embryonic stern cells.OBJECTIVE:To establish a feeder layer-and serum-free culture system of human embryonic stem cells.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Kunming Animal Institute of Chinese Academy of Sciences from September 2007 to February 2009.MATERIALS:Pregnancy 12.5 or 13.5-day ICR strain mica (clean grade) were provided by the Animal Center of Kunming Medical College.Human embryonic stem cell line BG02 was purchased from Bresagen,USA.METHODS:BG02 cells were digested,centrifuged,resuspended in feeder layer-free medium,and then incubated for 5-7 days.Differentiated human embryonic stem cells were removed.Dispase was added for digestion.Samples were cut into blocks,centrifuged,resuspended,and then incubated at 1:3 in a 4-well plate coated with laminin in feeder layer-free and serum-free medium,supplemented with 80% DMEM/F12,20% KSR,2 mmol/L glutamine,1% non-essential amino acid,0.1 mmol/L β-mercaptoethanol,insulin-transferrin-selenium (ITS) (×1),10~6 U/L penicillin,100 mg/L streptomycin,4 μg/L basic fibroblast growth factor (bFGF) and 0.12 μg/L transforming growth factor-β1 (TGF-β1).ICR fetus mica were sterilely obtained to culture mouse embryonic flbroblasts by tissue pancreatin digestion method.These cells were incubated in a 6-well plate coated with 0.1% gelatin.Human embryonic stem cell line BG02 cultured on mouse embryonic fibroblasts feeder layers was transferred to laminin-coated plates in serum-free medium containing bFGF,TGFβ1 and ITS.MAIN OUTCOME MEASURES:The morphology of BG02 cells in feeder layer-and serum-free condition was observed.The specific molecular markers of human embryonic stem cells were determined by immunohistochemical staining.The karyotype and differentiation ability in vitro of BG02 cells were analyzed.The differences in the proliferation and the differentiated rate of BG02 clumps in feeder layer-and serum-free condition or mouse embryonic fibroblast feeder layer condition were compared.RESULTS:BG02 calls had been continuously cultured for at least 20 passages in feeder layer-and serum-free culture condition.BG02 clones in this culture system had typical human embryonic stem cell morphology.BG02 cells all expressed SSEA-4,SSEA-3,TRA-1-60,TRA-1-81,Oct-4,but did not express SSEA-1.After 20 days of culture,BG02 cells had the capacity to differentiate into derivatives of embryonic endoderm,mesoderm,and ectoderm.Differentiated cells could express alpha-fetoprotein,nestin,α-actin.At passage 20,call karyotype was normal (46XY).The growth of BG02 cells cultured in feeder layer-and serum-free condition was more slowly compared with mouse embryonic fibroblast feeder,and doubling time was significantly prolonged (P ＜ 0.05).Differentiation rate of the colonies was significantly increased (P ＜ 0.05).CONCLUSION:A feeder layer-and serum-free condition for culture of BG02 cells has been established.The addition of bFGF,TGFβ1 and ITS can maintain the self-renewal of BG02 cells.

Objective To investigate the clinical efficacy and mechanism of vacuum-assisted closure (VAC) in the preoperative and postoperative treatment of bedsore united with skin flap.Methods Twenty two cases with bedsore were randomly divided into experimental and control groups.In the control group,the surgery of flap was performed after the treatment of continuous negative pressure about 7-10 days and the VAC was not applied after operation.While in the experimental group,VAC was not used before operation.It was applied on flaps as soon as sutured the border of flap and decubitus ulcers and removed after 7-10 days.By comparing the general appearance of two groups,microvessel count and the detection rate of bacterial culture and other indicators,the clinical effects of two treatments were investigated and the preliminary mechanism was analyzed.Results After preoperative VAC treatment,11 cases of control group showed a little granulation tissue growth,less subcutaneous hematoma and wound effusion,increased microvessel count and negative bacterial culture.However,there were 4 cases of death cavity residual,subcutaneous hematoma and wound effusion,positive bacterial culture and another 4 cases of delayed healing with skin flap repairing bedsore.The application of VAC in experimental group showed close contact of flap with the basement,less effusion,increased microvessel count and negative bacterial culture.One case of skin flap had a small area of separation,after the dressing of skin and the flap survived.The other wounds healed by first intention.Conclusions The use of VAC to repair bedsore can reduce the number of operation,and it is beneficial to the flap survival.

Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.

BACKGROUND: Whether human embryonic stem cells (hESCs) cultured on different feeder layers can maintain identical or similar characteristics remains unclear.OBJECTIVE: To compare the characteristics of hESCs cultured on human- and mouse- origin feeder layers.DESIGN, TIME AND SETTING: The in vitro cytology observation was performed at the Kunming Institute of Zoology, Chinese Academy of Sciences between September 2007 and February 2009.MATERIALS: Two ICR pregnant mice with 12.5-13.5 embryonic days were provided by Animal Center of Kunming Medical College. Immortalized human adult fibroblast (HAFi) cell line was presented by School of Medicine, Johns Hopkins University (USA). hESCs line was provided by Kunming Institute of Zoology, Chinese Academy of Sciences.METHODS: Mouse embryonic fibroblasts (MEF) were harvested from ICR mice by trypsinization method. HAFi were conventionally cultured. After γ ray treatment, two kinds of cells were incubated on 6-well gelatin-coated plate with density of 2.5×10~4/cm~3. hESCs were cultured on HAFi or MEF feeder cells containing β-mercaptoethanol DMEM/F12 and basic fibroblast growth factor.MAIN OUTCOME MEASURES: Morphology, expressions of specific molecular markers, Oct-4 positive rate, as well as cell doubling time of hESCs cultured by two methods were compared.RESULTS: ①BG02 cells on MEF and HAFi shared the similar morphology and characteristic pluripotency markers, which expressed SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4, but not SSEA-1. However, the proportion of positive Oct-4 cells in hESCs colonies maintained on MEF was lower than that of HAFi (P < 0.05) with shorter doubling time (P < 0.05).CONCLUSION: hESCs cultured on MEF and HAFi represent some differences in the growth and pluripotency characteristics.

Objective To study the metabolic characteristics in type 2 diabetes mellitus with different BMI.Methods 233 patients with type 2 diabetes mellitus were divided into 2 groups. The first group was the BMI ＜25kg/m2 ,the second group with BMI ≥25kg/m2. The blood sugar、 blood-fat 、blood uric acid、type-B ultrasonic of liver were analyzed. Results The waistline、hip circumference、limosis c-peptide 、total cholesterol 、triglyceride and morbidity of fatty liver and hypertension of the second group were higher than the first group. Conclusion Over weigh patients of type 2 diabetes had IR、hyperlipemia、hypertension 、fatty liver. The rate and the severity of the artery diseases in type 2 diabetes mellitus increased wit the increase of the BMI. So weigh reduction could decrease the development of the artery diseases in type 2 diabetes.

Objective To establish an efficient HPLC method for the determination of uric acid in plasma of hyperuricemia model mice,and the evaluation of uric acid lowering effect of Lesinurad.Methods The Laballiance Series Ⅲ HPLC system was adopted with Kromasil C18 column (100 mm × 4.6 mm,5 μm).The mobile phase consisted of methanol-0.5％ acetic acid (10:90) for isocratic elution with a flow rate of 0.4 mL/min.The detection wavelength was set at 283 nm.The established HPLC method was used to detect the plasma uric acid level of mice at 0.5,1.0,and 2.0 h time points after which being ip injected with 250 and 500 mg/kg uric acid.Lesinurad of 250 and 500 mg/kg was ig given to mice,0.5 h later,mice were ip injected with 500 mg/kg uric acid to establish hyperuricemia model,and 1 h later,the established HPLC method was used to detect the plasma uric acid level of mice.Results There was a good linear relationship between peak area and the concentration of plasma uric acid in the range of 7.5-150 μg/mL (r =0.997).The specificity,repeatability,precision,stability,and recovery of the established HPLC method was in accordance with the guiding rules of biological sample determination.Compared with the endogenous serum uric acid concentration of control group mice,serum uric acid concentration of 250 mg/kg dose group was significantly increased 0.5 h after ip administration with uric acid (P ＜ 0.01),and serum uric acid concentration of 500 mg/kg dose group was significantly increased 0.5,1.0,and 2.0 h after ip administration with uric acid.Compared with model group,the concentration of uric acid in plasma decreased significantly in low dosage group administered with Lesinurad (P ＜ 0.05),while decreased more significantly in high dosage group (P ＜ 0.01).Conclusion This convenient,rapid,and accurate method can be applied to the determination of uric acid in mouse plasma and the evaluation of relative drugs,which provide an efficient analysis way for establishing hyperuricemia model and screening relative drugs.

OBJECTIVETo observe the liver injury degree of SD rats after 28-day administration of aqueous extracts of Polygonum multiflorum (AEPM) and the correlation with cholestasis mechanism.

METHODAdult SD rats were orally administered with 30, 60 g x kg(-1) of APEM once every day for 28 d. After 28 d, the general condition of rats such as weight were observed, liver function-related indicators were detected. Bile was collected to determine total bile acid output, flow rate and density and changes in major compositions. Their livers were weighed then sent for histopathological examination.

RESULTAEPM did not change the general conditions and weights of rats. From the results of the related indicators of liver function and cholestasis, AEPM did not change the contents of ALT and AST in serum, but high dose of AEPM can increase the contents of ALP, GGT and TBA in serum (P < 0.05, P < 0.01) and decrease the content of TBIL in serum (P < 0.05). And the contents of GGT in serum of low dose rats were increased (P < 0.05). The bile flow was not changed by AEPM, but bile compositions of high dose male rats were obviously changed (TG increase, TBIL decrease, TBA decrease). The weights of liver and ratio of liver of the high dose rats were increased but showed no statistical significance. Pathologic examination displayed that there were only small pieces of necrosis in livers of several rats, without any severe disease.

CONCLUSIONAEPM can obviously injure bile duct epithelial cells, intervene liver cell functions and change bile compositions in rats, thus it is proved to induce cholestasis without severe liver injury.

Administration, Oral ; Animals ; Bile ; chemistry ; drug effects ; Cholestasis ; chemically induced ; Female ; Liver ; drug effects ; pathology ; Male ; Plant Extracts ; toxicity ; Polygonum ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective:To compare the clinical efficacy of platelet-rich plasma (PRP) injection and conventional treatment for partial rotator cuff tear and analyze the possible mechanism.Methods:A prospective randomize control study was performed for clinical data of 100 patients with partial rotator cuff tear admitted to Beijing Tongren Hospital affiliated to Capital Medical University between January 2019 and December 2020. The patients were assigned to observation group ( n=50) and control group ( n=50) according to the random number table. Both groups were given conventional non-operative treatment, together with PRP injection in observation group and steroid injection in control group. The visual analogue score (VAS), Constant-Murley Score (CMS) and serum levels of inflammatory cytokines [interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α)] were measured before and at 3, 6 months after treatment. The MRI images were also evaluated in observation group before treatment and at 6 months after treatment. Results:There were 55 males and 45 females with the age range of 40-75 years [(55.9±9.1)years]. All patients were followed up for 6.0-8.2 months [(6.2±2.9)months]. There were no significant differences between the two groups in VAS, CMS, IL-6, IL-1β and TNF-α before treatment (all P>0.05). The VAS was (3.5±0.9)points and (1.4±0.4)points] in observation group at 3, 6 months after treatment, significant lower than (4.1±0.9)points and (1.8±0.5)points in control group (all P<0.01). The CMS was (59.5±9.4)points and (86.5±7.9)points in observation group at 3, 6 months after treatment, significant higher than (53.5±8.6)points and (78.5±8.4)points in control group (all P<0.01). The serum levels of IL-6, IL-1β and TNF-α were (0.69±0.21)μg/L, (2.06±0.55)μg/L and (2.12±0.49)μg/L in observation group at 3 months after treatment, significant lower than (0.92±0.26)μg/L, (2.67±0.48)μg/L and (2.87±0.51)μg/L in control group (all P<0.01). The serum levels of IL-6, IL-1β and TNF-α in observation group [(0.37±0.11)μg/L, (1.14±0.23)μg/L, (1.27±0.25)μg/L] were also significantly lower than those in control group [(0.45±0.09)μg/L, (1.36±0.27)μg/L, (1.88±0.26)μg/L] at 6 months after treatment (all P<0.01). In both groups, the VAS, CMS and inflammatory cytokines (IL-6, IL-1β, TNF-α) showed significant improvement at 3, 6 months after treatment when compared to the pre-treatment levels (all P<0.05). Simutaneously, each group showed significant differences in each indicator at 3, 6 months after treatment ( P<0.05). In observation group, MRI showed that the high signal on the superior surface of the supraspinatus disappeared whereas the high signal in the tendon decreased at 6 months after treatment. Conclusion:For patients with partial rotator cuff tear, PRP injection can relive shoulder pain and improve shoulder function, which may be associated with the down-regulation of inflammatory responses.

Objective:To analyze the difference and reliability of blood 17-hydroxyprogesterone (17-OHP), an indirect screening index for congenital adrenal hyperplasia (CAH), between preterm and full-term infants.Methods:In this retrospective cross-sectional study, a total of 210 285 newborns who underwent CAH screening at the Neonatal Screening Center of Shanghai Children′s Hospital from January 2019 to December 2022 were collected, including 14 312 premature infants and 195 973 full-term infants.The concentration of 17-OHP in dried blood spots on filter paper was determined by an automatic fluorescence analyzer.The distribution of 17-OHP levels in preterm and full-term infants and its statistical index were analyzed.The Kolmogorov-Smirnov test was used for normal distribution.The skewed distribution data was converted into approximately normal distribution using Box-Cox.Outliers were eliminated by the interquartile range method.The cumulative frequency distribution map was drawn by R language programming.The 99.5 th percentile value was used as the screening threshold and compared with the reference value given by the manufacturer or laboratory and with the reference change value (RCV). Results:According to the threshold provided by the laboratory, 26.76‰ of premature infants were tested positive in preliminary screening, and 4 were confirmed with an incidence of 1∶3 578, while 0.79‰ of full-term infants were tested positive in preliminary screening, and 11 were confirmed with an incidence of 1∶17 816.The thresholds for CAH screening established indirectly were 20.35 nmol/L in preterm infants and 10.78 nmol/L in full-term infants.The relative deviations between the indirect CAH screening thresholds and the manufacturer′s or laboratory′s CAH screening thresholds were higher than the RCV, respectively.According to the indirect CAH screening thresholds, the negative and positive coincidence rates of 65 samples in 13 batches from the Centers for Disease Control and Prevention interlaboratory quality assessment program in the United States reached 100%.A retrospective analysis of 210 285 neonates showed that 17-OHP concentration was higher than the screening threshold in all CAH-positive neonates.The application of this screening threshold reduced the false positive rate of preterm infants by 59.79%.Conclusions:It is feasible to establish the CAH screening thresholds for premature and full-term infants by an indirect method, which can improve the efficiency of screening and provide better diagnostic basis for clinical practice.