In order to obtain an efficient screening procedure for identification of succinate producing anaerobic strains,a semi-quantitative paper chromatography method was developed. Lactic acid and acetic acid were identified as the main byproducts in the process of succinate production by the high performance liquid chromatography (HPLC).Succinic acid was completely separated from the byproducts of lactic acid and acetic acid in the same broth developed by paper chromatography.The content of succinic acid was calculated by a semi-quantitative method.The results showed that paper chromatography was a simple and cost effective method that could be utilized to screen anaerobic strains producing succinic acid.

Objective To explore the relationship between the serum level of E-selectin and S128R polymorphisms in the exon 4 of E-selectin gene and acute myocardial infarction (AMI) in local Han peoples. Method The genotype of E-selectin were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 168 patients with acute myocardial infarction and 200 healthy controls,and the serum level of E-selectin was measured by enzyme-linked immunosorbent assay (ELISA).Results There was significant difference in frequencies of allele and genotype in S128R polymorphism between acute myocardial infarction and control groups respectively,The relative risk suffered from acute myocardial infarction of SS genotype was 2.234 times of the SR genotype (OR=2.234,95% CI:1.112～4.437),The serum E-selectin level was significantly higher among carriers of SR genotype as compared with SS genotype (41.65?8.87)?g/L vs (34.23?6.72)?g/L,P

Objective To study the action of puerarin on Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and explore its mechanism. Methods SD female rats were randomly assigned into sham operation group, model group, premarin group and puerarin groups (120, 60, 30 mg/kg). The model group and sham operation group were injected NS intraperitoneally, other groups were treated with corresping drugs for 30 d. Immunohistochemistry and RT-PCR were used to determine Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons. Results Caspase-3 expression of hippocampal CA1 neurons was significantly decreased in high-dose puerarin group (P <0.05). Bcl-2 expression of hippocampal CA1 neurons was significantly increased in high- and medium-dose puerarin groups (P<0.05, P<0.01). Bcl-2 mRNA level of hippocampal CA1 neurons in high-dose puerarin group was significantly increased (P<0.05). Conclusion Puerarin can decrease Caspase-3expression and increase Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and has protective effect on neuronal structure.

OBJECTIVETo investigate the clinical characteristics and diagnosis of mucopolysaccharidosis VII.

METHODThe clinical and biochemical features of an infant with mucopolysaccharidosis VII confirmed by enzyme assay were analyzed.

RESULTThe 2 month-old male infant showed hydrops fetalis, mental retardation, coarse face, corneal clouding, hepatosplenomegaly, hernias, Alder-Reilly granules in the leucocytes and decreased platelet (32 × 10(9)/L). The biochemical markers showed urinary glycosaminoglycans (GAG) (532.8 mg/L, controls < 70.0 mg/L). The ratio of GAG/creatinine was 161.3 (controls: 26.2 ± 11.7). Serum chitotriosidase activity was 315.8 nmol/(ml·h) [control < 53 nmol/(ml·h)]. Beta-glucuronidase activity was deficient in isolated leukocytes.

CONCLUSIONSevere form of mucopolysaccharidosis VII exhibited characteristics of hydrops fetalis, hepatosplenomegaly, coarse face, thrombocytopenia and Alder-Reilly granules in the leucocytes. The measurements of GAG in urinary and beta glucuronidase in leucocytes are critical to diagnosis and deferential diagnosis.

Glucuronidase ; metabolism ; Glycosaminoglycans ; urine ; Humans ; Infant ; Leukocytes ; enzymology ; Male ; Mucopolysaccharidosis VII

Objective:To investigate the effect of small molecule antibacterial agent Halicin against Staphylococcus aureus. Methods:The minimum inhibitory concentration (MIC) and minimum bactericidal concentration of Halicin against S. aureus were detected by the microbroth dilution method. The time-kill assay of Halicin against S. aureus was detected by agar plate dilution method. Micro checkerboard dilution method was used to determine the synergistic antibacterial activity between Halicin and conventional antibiotics. Crystal violet staining method was used to assess the biofilm inhibitory and eradicating activity of Halicin. Hemolysis rate was used to detect the mammal cell toxicity of Halicin. Through the mouse skin abscess model, take the skin tissue around the abscess to grind and dilute the colony to detect the antibacterial effect of Halicin in vivo. Results:Halicin showed significant bacteriostasis effects against S. aureus with the minimum inhibitory concentration of 2-4 mg/L. Halicin could significantly reduce the average CFU counts of S. aureus about 5.5×10 6 CFU/ml in a concentration-dependent manner after 8 h treatment at the concentration of 16 mg/L. The fractional inhibitory concentration value between Halicin and ampicillin was 0.5, showing a synergistic antibacterial efficacy. Halicin effectively inhibited the formation of biofilms at the concentration of 4 × MIC, reducing the total biofilm biomass ( A570) from (2.89±0.09) to (1.35±0.17) ( t=11.12, P<0.05). However, there was no eradication effect against preformed biofilms. In addition, Halicin had almost no hemolytic activity on red blood cells even at the concentration up to 128 mg/L. It showed that 20 mg/kg Halicin reduced bacterial burden about 3.0×10 7 CFU/ml in vivo. Conclusion:Halicin had a strong antimicrobial activity against S. aureus with no hemolytic activity.

OBJECTIVETo investigate the co-culture of keratinocytes with Malassezia isolates which cause the pityriasis versicolor with different color and to analyze the changes of cytokines associated with melanogenesis.

METHODSThe effects of Malassezia species with different proportions on the growth rate of keratinocytes was assessed with 5 g/L methyl thiazolyl tetrazolium (MTT). Co-culture of keratinocytes and Malassezia species were performed with isolates from hyer- and hypo-pigmentation areas of pityriasis versicolor. The supernatants were collected at different time points, and the changes of basic fibroblast growth factor (b-FGF), endothelin-1 (ET-1), nerve growth factor-beta (NGF-beta), interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), stem cell factor (SCF) were recorded. Three control groups were established accordingly.

RESULTSWhen the ratio between keratinocytes and Malassezia species was lower than 1: 10, the growth rate of keratinocytes was not affected by Malassezia (P > 0.05). When the ratio was increased above 1:20, the growth rate of keratinocytes was significantly inhibited by Malassezia (P < 0.01). The secretions of IL-1alpha, IL-6, TNF-alpha, and ET-1 was significantly increased after the co-culture of keratinocytes and Malassezia (P < 0.01), while those of b-FGF, NGF-beta, and SCF had no significant changes (P > 0.05). Compared with the isolates from the hypo-pigmentation area, ET-1 induced by isolate from hyperpigmentation area significantly increased (P < 0.01).

CONCLUSIONWhen Malassezia isolates are co-cultured with keratinocytes, the secretions of cytokines associated with melanogenesis may differ from each other. ET-1 may play certain role in the hyper-pigmentation of pityriasis versicolor.

Cell Proliferation ; Cells, Cultured ; Cytokines ; biosynthesis ; Humans ; Keratinocytes ; cytology ; metabolism ; microbiology ; Malassezia ; isolation & purification ; physiology ; Melanins ; biosynthesis ; Tinea Versicolor ; microbiology

Objective:To compare the different follow-up methods on compliance and prognosis of non-selective beta blockers (NSBBs) for out-patients with secondary prevention of esophagogastric variceal bleeding (EGVB).Methods:A total of 72 patients with portal hypertension and esophageal gastric varices (EGV), who were admitted to the department of gastroenterology, Renmin Hospital of Wuhan University from July 2018 to April 2019 were randomly included in the traditional outpatient follow-up group (group A, 24), remote platform follow-up group (group B, 23) and combined follow-up group (group C, 25) according to the random number table. The patient′s medication compliance, heart rate response, condition change and treatment were recorded. NSBBs medication compliance, influencing factors and prognosis of patients in different groups were analyzed.Results:Forty (40/72, 55.6%) patients had good compliance with NSBBs, among which 34 (34/40, 85.0%) achieved heart rate response. The medication compliance of the group B (15/23, 65.2%) and the group C (17/25, 68.0%) was higher than that of the group A (8/24, 33.3%; χ2=4.778, P=0.029; χ2=5.889, P=0.015, respectively). There was no significant difference in the medication compliance between the group B and the group C ( χ2=0.042, P=0.838). In the group A, the subgroup of good compliance had higher proportion of local patients than that of poor compliance subgroup [7/8 VS 37.5% (6/16), P=0.033]. Patients with good drug compliance had higher proportion of gastroscopy review than that of poor compliance patients [75.0% (30/40) VS 21.9% (7/32), χ2=20.085, P<0.001] and less patients with EGVB [5.0% (2/40) VS 21.9% (7/32), P=0.073]. Conclusion:Patients with portal hypertension and EGV have poor compliance with NSBBs. Remote platform follow-up is a better way to improve compliance of drug prevention. Patients with good NSBBs compliance have a higher compliance of gastroscopy review. The risk assessment of variceal bleeding and endoscopic sequential therapy based on the results of gastroscopy review are expected to reduce the risk of EGVB.

OBJECTIVETo explore the incidence of various types of mucopolysaccharidosis (MPS) and their clinical characteristics.

METHODSA total of 75 children highly suspected as having MPS underwent quantitative and electrophoretic analysis of urinary glycosaminoglycans (GAGs) and enzymatic analysis of seven types of MPS from January 2009 to December 2011. Fluorescence assay was used to measure the activities of α-L-iduronidase, iduronate-2-sulfatase, α-N-acetylglucosaminidase, galactosamine-6-sulfatase, β-galactosidase, arylsulfatase B and β-glucuronidase in the white blood cells.

RESULTSA total of 52 cases were confirmed with MPS based on clinical, radiological, and enzymatic examinations. The 52 cases, with a mean age of 4.0 ± 2.2 years, included 5 cases of MPS I (10%), 20 cases of MPS II (38%), 20 cases of MPS IVA (38%), 6 cases of MPS VI (12%) and 1 case of MPS VII (2%). No MPS IV B cases or MPS IIIB cases were found. Compared with healthy children of the same age, the GAG/Cr ratio was significantly elevated in 50 confirmed cases of MPS (two MPS IVA cases having no increased ratio). All children with increased urinary GAGs had a confirmed diagnosis of MPS. The age of onset was between 1 and 2 years after birth in most cases, and often complicated by hernia and valvular heart disease. Children with MPS I, MPS II, and MPS VI presented with ugly and unsmooth face, short stature, joint stiffness, and limitation of motion, while children with MPS IVA presented with short stature, skeletal dysplasia, and joint laxity.

CONCLUSIONSType IVA and type II are the most common in MPS cases, followed by type VI and type I. MPS children are characterized by special appearances including ugly and unsmooth facial appearance, short stature and skeletal dysplasia. Quantitative analysis of urinary GAG, as a simple, rapid, and reliable method, is recommended for screening of MPS.

Acetylglucosaminidase ; blood ; Child ; Child, Preschool ; Creatinine ; urine ; Female ; Glucuronidase ; blood ; Glycosaminoglycans ; urine ; Humans ; Iduronidase ; blood ; Infant ; Magnetic Resonance Imaging ; Male ; Mucopolysaccharidoses ; diagnosis ; enzymology ; pathology ; beta-Galactosidase ; blood

Objective:High-throughput screening to obtain small molecular compounds against Gram-negative bacilli by targeting BamA outer membrane protein.Methods:The sybyl-X2.1 software was used to perform high-throughput virtual screening of small molecular compounds in Chemdiv compound library based on the molecular docking. The top 150 hits by high-throughput screening were re-screened through in vitro biological experiments. The top 4 small molecules with obvious antibacterial activity were selected for in-depth molecular docking analysis, and the small molecule 8308-0401 with the highest docking score was selected for further experiments. The antibacterial effect of 8308-0401 combined with rifampicin was tested by checkerboard assay. Finally, the affinity between 8308-0401 and BamA was tested by plasma surface resonance assay. Results:The docking score of the top 150 hits calculated by high-throughput virtual screening had a mean value of 5.63. In vitro biological experiments showed that small molecules 8308-0401, 8365-1335, C066-2507 and L582-0346 exhibited strong antibacterial activity. Among those molecules, 8308-0401 showed the highest molecular docking score, and synergistic antibacterial activity against both types of strains and clinical isolates when combined with rifampicin. 8308-0401 has a strong affinity to BamA with binding a constant of 182 μmol/L. Conclusion:The small molecule 8308-0401 exerts antibacterial activity against Gram negative bacilli by targeting the outer membrane protein BamA.

OBJECTIVETo explore the relationship and interaction of elevated fasting glucose and hypertension on cardiocerebral vascular disease.

METHODS10 054 males and females were recruited for our cross-sectional study during May 2007 to August 2007. Unconditional logistic regression was used to analysis the relationship between fasting glucose and hypertension on cardiocerebral vascular disease. A product of fasting glucose and hypertension was added to the logistic regression model to evaluate the multiplicative interaction and relative excess risk of interaction (RERI), attributable proportion (AP) of interaction and synergy index (S) was applied to evaluate the additive interaction of the two factors. Bootstrap was used to calculate 95% confidence intervals (CI) of RERI, AP and S.

RESULTSAfter adjusting age, gender, smoking, drinking, body mass index (BMI) and region, the product of fasting glucose and hypertension was not statistically significant, which means there was no multiplicative interaction between the two. But the additive indexes RERI, AP and S with 95%CI of diabetes and hypertension were 0.64 (0.03, 1.25), 0.27 (0.01, 0.47) and 1.83 (1.02, 5.13) respectively, which means significant additive interaction was shown between the two on cardiovascular disease but not no stroke. And there were no additive interaction between impaired fasting glucose on cardiovascular disease or stroke.

CONCLUSIONSHypertension was independently related to cardiovascular disease and stroke in Beijing citizens, and diabetes were independently related to stroke. There was additive interaction between diabetes and hypertension on cardiovascular disease.

Adult ; Aged ; Blood Glucose ; metabolism ; Blood Pressure ; Cardiovascular Diseases ; epidemiology ; Cerebrovascular Disorders ; epidemiology ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Hypertension ; blood ; complications ; Male ; Middle Aged ; Risk Factors