Objective To detect the creatin kinase (CK) and its isozeyme (CK-MB) activity in serum of the healthy black bear (Ursus thibetanus) reared in pens to establish reference standard of biochemistry for CK and CK-MB activities of black bears as well as to offer scientific basis for prevention and cure of the black bear disease. Methods The CK activity and CK-MB activity were measured using spectrophotometer based on the principle of enzymatic reaction rate. All data were used to calculate the mean value, variation coefficient and the confidential interval of expected value of enzymatic activity. Results Under 30 ℃, of the male black bears, the average CK activity is 162.70 U/L, the variation coefficient 11. 90%, the confidential interval of expected value 148.90 to 176.50 U/L, and the average CK-MB activity is 17.4 U/L, the variation coefficient 17%, the confidential interval of expected value 15.26 to 19.54 U/L. Of the female black bears, the average CK activity is 149.3 U/L, the variation coefficient 8%, the confidential interval of expected value 140.68 to 157. 92 U/L, and the average CK-MB active is 15.8 U/L, the variation coefficient 18.6%, the confidential interval of expected value 13.70 to 17.90 U/L. Conclusion The data we obtained can serve as biochemical indexes for the healthy black bears.

Objective To observe the prevalence of hyperuricemia(HUA)among obese children,and to ex-plore the association between uric acid(UA)levels and cardiometabolic risk factors,acanthosis nigricans and non - al-cohol fatty liver disease(NAFLD). Methods By using representative sampling method,1 753 obese children aged 6 -17 years old from 18 schools in 3 districts of Beijing(Xicheng,Haidian,Miyun)were selected to participate in the clini-cal examinations,including anthropometric measurements(height,weight)and blood pressure. Serum biochemical pa-rameters were assessed,including fasting plasma glucose(FPG),total cholesterol(TC),triglyceride(TG),high - densi-ty lipoprotein cholesterol(HDL - C),low - density lipoprotein cholesterol(LDL - C)and UA. Acanthosis nigricans and B - model ultrasonography of the liver were conducted. Results The prevalence of hypertension,impaired fasting glu-cose,dyslipidemia,acanthosis nigricans,and NAFLD among these 1 753 obese children was 33. 6%(589 cases), 66. 5%(1 156 cases),54. 3%(943 cases),23. 3%(408 cases),and 17. 0%(298 cases),respectively. The preva-lence of HUA was 40. 70%(714 / 1 753 cases),with 50. 17%(581 / 1 158 cases)in boys and 22. 34%(133 / 595 ca-ses)in girls. There was a significant increase in body mass index,systolic blood pressure,diastolic blood pressure, FPG,TG and LDL - C with the increase of UA,but there was a decrease in HLD - C with the increase of UA(all P ﹤0. 05). In boys,the adjusted odds ratios( OR)and 95% CI of the highest quartile of UA for hypertension,impaired fasting glucose,dyslipidemia,acanthosis nigricans,and NAFLD were 1. 16(0. 77 - 1. 74),1. 34(0. 90 - 1. 99),1. 29 (0. 89 - 1. 87),1. 89(1. 17 - 3. 04),and 1. 71(1. 03 - 2. 84),respectively;in girls,the adjusted OR and 95% CI of the highest quartile of UA for hypertension,impaired fas-ting glucose,dyslipidemia,acanthosis nigricans,and NAFLD was 0. 70(0. 40 - 1. 24),0. 60(0. 40 - 1. 00),1. 69(1. 04 - 2. 70),1. 67(0. 80 - 3. 49),and 1. 33(0. 48 - 3. 66),re-spectively. Conclusions The prevalence of HUA is relatively high in obese children and there is a strong association between UA and some car-diovascular metabolic disorders,acanthosis nigricans and NAFLD.

This study was performed to investigate the therapeutic effect of oroxylin A(OA)on chicken type Ⅱcollagen-induced arthritis(CIA)in mice and observe the changes of immune cells.The CIA model was established,and 40 mg/kg OA was intraperitoneally injected for 10 consecutive days from the 28th day.The mice were sacrificed at three different times during the administration period,and the joints were scored at each time point.HE staining was used to observe the pathology of the mouse ankle joint;flow cytometry was used to detect the changes of Th17,Treg and macrophages in spleen and inguinal lymph nodes;ELISA was employed to detect the expression levels of IL-1β,IL-18 and IL-6 in serum;and qRT-PCR was used to detect the expression levels of IL-1β,IL-18,IL-6 and TNF-β in spleen of mice.Data showed that on the 34th day after OA administration,the joint swelling of CIA mice was significantly relieved,the pathological score was decreased,and the inflammatory cell infiltration was decreased.Flow cytometry results showed that the proportion of Th17 cells and macrophages in the spleen and inguinal lymph nodes of CIA mice in OA group decreased,while the proportion of Treg cells increased.The results of ELISA and qRT-PCR showed that OA could inhibit the level of inflammation in CIA mice.In conclusion,OA can regulate the proportion of immune cells,inhibit the level of inflammation in CIA mice,and then relieve the symptoms of CIA mice.

OBJECTIVE:To investigate the effects of UGT1A4142T>G polymorphism and blood concentration of valproic ac-id(VPA)on blood concentration of lamotrigine(LTG)in southern Chinese Han children with epilepsy,and to establish the predic-tion equation for quantitatively estimating blood concentration of LTG. METHODS:A total of 72 southern Chinese Han children with epilepsy selected from Guangzhou Women and Children's Medical Center during Jan. 2010-Sept. 2016 were given LTG+VPA. LC-MS/MS and enzyme amplified immunoassay were adopted to determine the blood concentration of LTG and VPA. RFLP-PCR was adopted to determine UGT1A4142T>G polymorphism. The relationships of age, gender, blood concentration of VPA, UGT1A4142T>G polymorphism and LTG concentration-to-dose-ratio (CDR) were also investigate. The prediction equation for blood concentration of LTG was established by multiple linear regression analysis. RESULTS:Age and blood concentration of VPA were positively related to CDR of LTG(r=0.225,0.300,P<0.05);there was no statistical significance in the influence of gender on CDR of LTG(P>0.05). UGT1A4 TT,TG and GG genotypes were detected in 39,29,and 4 cases respectively;the frequencies of each genotype were in line with the Har-dy-Weinberg balance(P>0.05). CDR of LTG of TT genotype was significantly lower than those of TG and GG genotype,with sta-tistical significance(P<0.05). Results of multiple linear regression analysis showed that the dose of LTG(x1),body weight(x2), blood concentration of VPA(x3)and UGT1A4142T>G polymorphism(x4)were all related to blood concentration of LTG(P<0.05). Using blood concentration of LTG(c)as dependent variable,above factors as independent variable,the regression equation was c=0.794+0.032x1-0.057x2+0.010x3+0.532x4(R2=0.616,P<0.05;UGT1A4 TT genotype was equal to 0,TG and GG genotype was equal to 1). There was a strong positive correlation between predicted blood concentration and measured ones(r=0.785,P=0.001). CONCLUSIONS:The dose of LTG,body weight,blood concentration of VPA and UGT1A4142T>G polymorphism may associated with blood concentration of LTG. Established prediction equation can provide reference for precise medication in south-ern Chinese Han children with epilepsy.

Objective To investigate the effects of KRAS mutation isoforms on tumor regression grade(TRG)of rectal cancer received neoadjuvant therapy.Methods The clinicopathologic data of patients with locally advanced and metastatic rectal cancer(stage Ⅲ-Ⅳ)who underwent radical tumor resection after neoadjuvant therapy were collected.The correlations between the mutated subtypes of KRAS and TRG as well as the clinicopathological features were analyzed.Results A total of 95 patients were enrolled,including 55 patients with poor tumor regression.The incidences of G12V mutation in exon 2 and A146 mutation in exon 4 of the KRAS gene were higher in the group with poor tumor regression than those in the significant tumor regression group(P＜0.05).In patients with G12D mutation of KRAS gene,the histological grade of tumor in poor tumor regression was higher than that in significant tumor regression group(P=0.027).In patients with poor tumor regression,the levels of CD8+,PD-1+T-lymphocyte infiltration were higher in G12D mutation subgroup than those in G12V mutation and G13D mutation subgroups(P＜0.05).Conclusions KRAS G12V mutation and A146 mutation suggest poor neoadjuvant therapy effect for rectal cancer;for patients with KRAS G12D mutation and poor tumor regression,the potential beneficiary for immunotherapy would be screened by detecting CD8+and PD-1+T-lymphocyte infiltration.

Objective To investigate the effect of age,gender,weight and UGT1A4142T>G gene poly-morphism on the efficacy of LTG in epileptic children treated with valproic acid ,and to determine the effective se-rum concentration of LTG in children with epilepsy in south China. Methods A total of 72 pediatric patients with epilepsy received LTG and VPA treatments were enrolled in this study. Patients were treated from January 2010 to September 2016 in Guangzhou women and childrens′medical center. Serum concentration of LTG was measured by using the liquid chromatography-tandem mass spectrometry. UGT1A4142T > G was genotyped by the polymerase chain reaction-restriction fragment length polymorphism method. The correlations between the efficacy of LTG and age,gender,weight were analyzed by chi-square test,non-parametric test and logistic regression analysis,respec-tively. Results The curative effect of patients who were younger and with lighter weight were relatively poor ,and men were better than women in the curative effect. UGT1A4142T > G was not related with LTG efficacy. When combined with VPA,the effective serum concentration of LTG in children with epilepsy was more than 2 g/mL. Conclusion There is a good correlation between age and LTG curative effect. The effective serum concentration of LTG in children with epilepsy,who were co-treated with VPA,was more than 2 g/mL. This study provides a refer-ence for the individual administration of children with epilepsy in south China.

Objective To investigate the effect of restrictive intravenous fluid on the complications and quality of life in the advanced hepatocellular carcinoma patients.Methods Clinical data of Three hundred and fifty-seven cases with advanced hepatocellular carcinoma from Mar 2010 to Mar 2016 was retrospectively analyzed.One hundred and sixty-eight cases were recruited in the restrictive intravenous fluid (RIF) group,and One hundred and eighty-nine cases were involved in the control group.The average volume of intravenous fluid of each day,plasma albumin concentration,splanchnocoel hydrops rate,phlebitis,incidence of vomiting,cancer related pain degree,anxiety degree were compared in the two groups.Results The average volume of intravenous fluid of each day in the RIF group [(720.29 ± 106.84) ml] were much lower than that in the control group [(1 820.36±342.12)ml] (P ＜0.05).The plasma albumin concentration in the RIF group [(35.65 ± 2.21)g/L] were higher than that in the control group [(32.25 ±2.32)g/L] (P ＜0.05).The rate of splanchnocoel hydrops,phlebitis,vomiting,bedsores,and hypstatic pneumonia in RIF group were 6.25％,4.69％,8.59％,3.9％,11.72％ and those in the control group were 13.97％,10.92％,17.47％,10.04％,and 24.45％,respectively (P ＜ 0.05).Moreover,the scores of cancer related pain and anxiety were much lower in the RIF group (5.21 ± 1.09,39.12 ± 5.54) than those in the control group(5.68 ± 1.18,41.56 ± 6.78) (P ＜ 0.01).Conclusions Restrictive intravenous fluid therapy can decrease the cancer associated complications and improved the quality of life in the advanced hepatocellular carcinoma patients.

Objective:To compare the clinical characteristics and analyze the prognostic factors between human immunodeficiency virus (HIV)-infected patients and non-HIV-infected immunocompromised patients with pneumocystis pneumonia (PCP) complicated with acute respiratory failure (ARF) in intensive care unit (ICU).Methods:The clinical data of patients with PCP complicated with ARF admitted in ICU of The First Affiliated Hospital of Zhengzhou University and The Sixth People′s Hospital of Zhengzhou City between May 2018 and October 2020 were retrospectively reviewed. All subjects were divided into HIV-infected group and non-HIV-infected immunocompromised group. General characteristics and underlying diseases of patients in the two groups were analyzed. Laboratory parameters, treatment and outcomes between two groups were compared. Independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis, and univariate and multivariate logistic regression models were used to identify the risk factors for the clinical outcome. Results:A total of 129 PCP complicated with ARF patients were enrolled, including 75 HIV-infected patients and 54 non-HIV-infected immunocompromised patients. Only 10.7%(8/75) patients of HIV-infected group received anti-retroviral therapy (ART), but none of the patients in either groups had previously received trimethoprim-sulfamethoxazole (TMP-SMX) for PCP prophylaxis. Acute physiology and chronic health evaluation (APACHE) Ⅱ score of HIV-infected group was 18.7±6.0, which was higher than that in non-HIV-infected immunocompromised group (13.1±4.4) when admitted in ICU ( t=-5.45, P<0.001). Hypoproteinemia was common in both groups. Ninety-six percent (72/75) of HIV-infected patients had CD4 + T lymphocyte counts lower than 200/μL and 84.0%(63/75) of patients had CD4 + T lymphocyte counts even lower than 50/μL, while 5.74%(31/54) of patients in non-HIV-infected immunocompromised group had CD4 + T lymphocyte counts lower than 200/μL. The CD4 + /CD8 + T lymphocyte counts ratio was 0.05(0.02, 0.12) in HIV-infected group, which was lower than that in non-HIV-infected immunocompromised group (0.96(0.64, 1.44)), and the difference was statistically significant ( Z=-9.16, P<0.001). The length of ICU stay and hospital stay of non-HIV-infected immunocompromised patients were 10.0(7.0, 14.0) days and 18.0(11.8, 32.5) days, respectively, which were both longer than those in HIV-infected patients (7.0(4.0, 9.0) days and 13.0(7.0, 23.0) days, respectively), and the differences were both statistically significant ( Z=-3.58 and -2.73, respectively, both P<0.050). The hospital mortality of HIV-infected patients was 57.3%(43/75), which was significantly higher than that in non-HIV-infected immunocompromised patients (38.9%, 21/54) ( χ2=4.27, P=0.039). Multivariable logistic regression identified that lactic dehydrogenase (LDH), C-reactive protein (CRP) and APACHE Ⅱ score were the risk factors for the clinical outcome of HIV-infected patients (odds ratio ( OR)= 1.006, 1.015 and 1.736, respectively, all P<0.050). The partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen (PaO 2/FiO 2), LDH and CD4 + T lymphocyte counts were the risk factors for the clinical outcome of non-HIV infected immunocompromised patients ( OR=0.970, 1.008 and 0.989, respectively, all P<0.050). Conclusions:PCP patients with ARF are critically ill with high mortality rate. LDH, CRP and APACHEⅡscore are predictors for prognosis of HIV-infected patients with PCP, while PaO 2/FiO 2, LDH and CD4 + T lymphocyte counts are predictors for prognosis of non-HIV infected immunocompromised patients with PCP.

Background and Objectives: Systemic scleroderma (SSc) is a rare and serious connective tissue disease, an autoimmune disease, and a rare refractory disease. In this study, preventive effect of single systemic human umbilical cord mesenchymal stem cells (UC-MSCs) transfusion on SSc was preliminarily explored. Methods: and Results: SSc mouse model was established by daily intradermal injection of Hypochlorite (HOCl). SSc mice were treated by single transfusion of UC-MSCs at 0.625×10 5 , 2.5×105 and 1×106 respectively. At the 42nd day of intradermal injection of HOCl, the symptoms showed up by skin and alveolar wall thickening, lymphocytic infiltration, increased collagen in skin/lung, and the increased proportion of CD3 ＋ CD4＋ CD25＋ FoxP3＋ cells (a Treg subset) in spleen. After UC-MSCs transfusion, the degree of skin thickening, alveolar wall thickening and lymphocyte infiltration were decreased, the collagen sedimentation in skin/lung was decreased, and the proportion of CD3＋ CD4＋ CD25＋FoxP3＋ cells was decreased. Conclusions UC-MSC can achieve a preventive effect in SSc mice by fibrosis attenuation and immunoregulation.

Objective:To investigate the molecular mechanism of VRC01 resistance in HIV-1 subtype B′ strains isolated from a patient (DRVI01) with broadly neutralizing antibody (bNAb).Methods:Sequences of the HIV-1 subtype B′ strains isolated from patient DRVI01 were compared with those of HIV-1 subtype B′ strains that were isolated at the same time but sensitive to VRC01 antibody. Key amino acids that might affect the neutralization of VRC01 were selected according to literature reports. Effects of the selected amino acids on VRC01 neutralization were verified by site-directed mutation and sequence exchange of membrane proteins from different patients.Results:Single-point mutations of E279D and R282K in LoopD region and N460A and N463Q in V5 region reversed the viral sensitivity to VRC01 neutralization. Combined mutations in two or three above-mentioned sites significantly increased the viral sensitivity to VRC01 antibody compared with single-point mutations. Contrary to literature reports, the glycosylation site mutation of N276 had no influence on the viral sensitivity to VRC01.Conclusions:HIV-1 subtype B′ strains isolated from patient DRV01 with bNAb carried the mutations of D279 and K282 in LoopD region and N460 and N463 in V5 region, resulting in resistance to VRC01 antibody.