Objective To establish a RP-HPLC method for determining the concentration of prulifloxacin active metabolite in human plasma and urine.Methods The supernatant obtained by centrifugation after the sample was precipitated with methanol- acetonitrile (1:1) was chromatographically separated on a Diamonsil C_(18)(250 mm?4.6 mm,5?m) using a mobile phase con- sisting of acetonitrile and 0.05 mol/L potassium dihydrogen phosphate (pH2.2) containing 1% tetrabutylammonium bromide. The solutions of 20:80 (V/V) and 12:88 (V/V) at a flow rate of 1.0 mL/min and 1.6 mL/min were used for plasma and u- rine, respectively.Then the samples were assayed at wavelength of Ex 280 nm and Em 425 nm.Results The linear range for prulifloxacin active metabolite in plasma and urine were 0.005-5 mg/L (r=0.9999) and 0.05-5 mg/L(r=0.9999)with a low- er limit of quantitation of 0.002 mg/L and 0.01 rag/L, respectively.In plasma, the relative recovery ranged from 100.64% to 101.00% at the concentration of 5.00, 0.50 and 0.05 mg/L and within-day and between-day precisions were less than 2.5% and 4.6% respectively.Meanwhile, the relative recovery ranged from 97.20% to 100.20% at the concentration of 2.50, 0.50 and 0.10 mg/L in urine.The within-day and between-day precisions were lower than 1.3% and 4.3%, respectively.The method had been successfully used for the pharmacokinetic studies of a prulifloxacin formulation after oral administration to healthy volunteers.Conclusions The present method is simple, rapid, accurate, reproducible and suitable for the pharmacoki- netic study of prulifloxacin in humans.

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

Abstract: Objective　To explore the spatial epidemiological characteristics of severe cases hand, foot and mouth disease (HFMD) in Guangxi, China, from 2014 to 2018, and to provide a basis for identifying the high-risk regions as well as the prevention and control of severe cases of HFMD in Guangxi. Methods　Spatial-temporal scanning analysis, global and local spatial autocorrelation analysis were used to analyze the spatial clustering of HFMD. The trend surface analysis was used to evaluate the spatial distribution trend of HFMD. Results　From 2014 to 2018, the incidence and severe case fatality rates of HFMD were 3.89/100 000 and 4.23%, respectively. Monte Carlo scanning analysis showed that the first cluster region was Cenxi City, the second cluster was mainly concentrated in northwest of Guangxi, and the aggregation time was mainly concentrated in April to May and August to October. The global spatial autocorrelation analysis showed that the severe HFMD was significant clustering distribution, and the Moran's I coefficients of the sever cases, severe morbidity and severe case fatality rate were 0.088, 0.118, 0.197, respectively (P<0.05). Local spatial autocorrelation analysis showed that hotspots of severe HFMD cases were concentrated in the southern Guangxi, mainly in Lingshan County. Anselin local Moran's I clustering and outlier analysis indicated that 5 high-high (H-H) clustering regions for fatality were Lingshan, Pubei, Zhongshan, Zhaoping and Pinggui County. There were 6 high-high (H-H) clustering regions for severe incidence rate, namely Lingshan, Qinnan, Lingyun, Youjiang, Bama Yao Autonomous and Pinggui County, and 1 high-low (H-L) clustering region, Cenxi County. The trend surface analysis showed that the overall number of severe cases of death decreased from east or west to the middle, and increased from north to middle, and then decreased to south. Conclusions　Severe HFMD cases in Guangxi have obvious spatial-temporal clustering, and the hop spots are mainly concentrated in southern Guangxi. The prevention and control of HFMD in areas with high incidence of severe cases should be strengthened to reduce the burden of HFMD cases.

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

On the basis of MEDLINE and EMBASE database, through bibliometrics, the quantitative research was conducted on the published literatures on the acupuncture-moxibustion clinical trial abroad. The situation of published articles in each continent, country and institution was analyzed statistically. It was found that the number of published articles was higher in Germany, America, England, Sweden, Austria, Japan, South Korea, etc. In Europe, the clinical trial of acupuncture and moxibustion was in the tendency of more country participants, wider distribution and larger amount of research. In North America, America was the main country for the study. In Asia, Japan and South Korea played the leading role. Of those countries, some institutions in Germany America, and South Korea were on the top of the list. In future, the above-mentioned countries and institutions should be monitored specifically so as to launch the active cooperation and strategic project.
Acupuncture Therapy ; Asia ; Bibliometrics ; Clinical Trials as Topic ; Europe ; Humans ; Moxibustion ; United States

Objective To explore the impact and regularity of snail population after changing of water level and to develop effective control and prediction programs. Methods A typical snail habitat closed to Changjiang county in Yueyang city, Hunan province was selected as the survey field.Data on water level and the changing trend of snail population during 2001-2009 including the average water level, maximum and minimum water levels, snail and infected snail densities,proportion of sampling frames with living and infected snails etc. were collected. The distribution of water level and snail indexes were described and a general additive model(GAM)for the relationships between these indexes were also fitted. Results Impacts of the average water level and the watered-out days in current year in the surveyed field on the snail density were statistically significant(P＜0.01), showing a quadratic curve association. The snail density remained at the lowest level when the average water level was at 25.0 m and the surveyed field watered-out days was 120. The average water level in current year and the field watered-out days in the past year showed statistically significant impacts on the infected snail density(P＜0.001), the proportion of sampling frames with infected snails(P＜0.05)and living snails(P value neared 0.05), presented a quadratic curve, a cubic curve or even a more complicated piecewise curve association. Both the infected snail density and the proportion of sampling frames with infected snails remained at the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 110, while the proportion of sampling frames with living snails kept the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 160. Conclusion The water level and the field watered-out days affected the development of snail population directly. The changing water level had an impact on snail population change, which mainly presented as nonlinear smooth function relation. Impact of the field watered out days on the infected snail density and the proportion of sampling frames with living snails and infected snails showed a hysteresis effect. The snail density was predicted to be retaining a high level when the water level was 24.0 m and the field watered-out days was 3 months. It had obvious advantages to fit the relationship of the changing water level and the snail indexes with a GAM which could get closer to the reality as well as easier to find and explain the potential associations and regulations.

OBJECTIVETo explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.

METHODSHepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and

NAD(P)Hquinone oxidoreductase-1(NQO1) were analyzed by Western blot.

RESULTSCompared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).

CONCLUSIONThe in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.

Antioxidant Response Elements ; Culture Media ; Fatty Acids, Nonesterified ; Fatty Liver ; metabolism ; GA-Binding Protein Transcription Factor ; Glutathione Peroxidase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Malondialdehyde ; metabolism ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism ; Nitric Oxide ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; metabolism

AIMTo detect the deletion distribution of dystrophin gene and dystrophin changes in muscle cells of the patients with Duchenne/Becker muscular dystrophy (DMD/BMD), furthermore to investigate the relationship between them and clinical symptoms.

METHODS42 patients with DMD/BMD were screened by 9 primers multiplex PCR. The patients from 5 DMD and 2 BMD were detected by immunofluorescence technique for analyzing dystrophin located in muscle cell membrane, compared with 2 normal males.

RESULTSThe deletion of one or more exons was found in 21 patients. 16 cases (76.2%) were detected in the central region and 5 patients (23.8%) in the 5' extreme region, especially in exon 48 (6 patients). Negative result of staining was seen in 5 DMD patients. Of these, one case of DMD had no detectable levels of dystrophin, but no deletion of DMD gene. Dystrophin immunostaining from two BMD patients consisted of a discontinuous staining pattern around most fibers.

CONCLUSIONIt might be possible that some correlation existed between the type of gene deletion and the degree of severity of the disease. The amount and size of exon deletion may not affect the symptoms. DMD/BMD are highly heterogeneous in clinical manifestation and in inheritance pattern. The pathologic foundation of DMD and BMD is the absence or abnormal expression of dystrophin. The consequence of that depends not only on the degree, but also on the function.

Adolescent ; Adult ; Child ; Child, Preschool ; Dystrophin ; analysis ; Exons ; Gene Deletion ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; genetics ; Sequence Deletion ; Young Adult

OBJECTIVEOur previous work indicated that overexpression of imprinting gene PEG10 is associated with malignant phenotype of hepatocellular carcinoma. The aim of this study is to explore whether disregulation of PEG10 leads to dysregulation of microRNAs.

METHODSIn silico analysis using TargetScan indicated that miR-122 could regulate the expression of PEG10. The expression of miR-122 in three hepatoma cell lines, Huh7, Hep3B and HepG2 and in primary human normal liver cell were compared using real time RT-PCR. After pre-miR-122 was transfected into HepG2 cell, the levels of PEG10 mRNA and protein were measured.

RESULTSIn silico analysis revealed that miR-122 could regulate the expression of PEG10. Real time RT-PCR indicated that miR-122 was not expressed in Hep3B and HepG2 cells, and only weakly expressed in Huh7 cells, but highly expressed in primary human normal liver cells. The expression of miR-122 was negatively correlated with the expression of PEG10. After pre-miR122 was transfected into HepG2, the mRNA level of PEG10 was not increased, whereas the protein level of PEG10 was increased.

CONCLUSIONmiR-122 may be involved in regulation of PEG10 expression.

Carcinoma, Hepatocellular ; genetics ; Gene Expression Regulation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; MicroRNAs ; genetics ; Proteins ; metabolism

BACKGROUND: Hypothermia is associated with many adverse clinical outcomes in pediatric patients, and thus, it is important to find an effective and safe method for preventing peri-operative hypothermia and its associated adverse outcomes in pediatric patients. This study aimed to investigate the effect of forced-air warming blankets with different temperatures on changes in the transforming growth factor-β (TGF-β), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-10 levels in children undergoing surgical treatment for developmental displacement of the hip (DDH). METHODS: The study included 123 children undergoing surgery for DDH under general anesthesia. The patients were randomly assigned to three groups, using a random number table: the 32, 38, and 43°C groups according to the temperature setting of the forced-air warming blankets. For each patient, body temperature was recorded immediately after anesthesia induction and intubation (T0), at initial incision (T1), at 1 h after incision (T2), at 2 h after incision (T3), at the end of surgery (T4), immediately upon return to the ward after surgery (T5), and then at 12 h (T6), 24 h (T7), 36 h (T8), and 48 h (T9) after the surgery. The serum levels of TGF-β, TNF-α, IL-1β, and IL-10 were measured at T0 and T4 for all groups. RESULTS: The number of patients with fever in the 38°C group was significantly less than those in the 32 and 43°C groups (χ = 6.630, P = 0.036). At T0, the body temperatures in the 38 and 43°C groups were significantly higher than that in the 32°C group (F = 17.992, P < 0.001). At T2, the body temperature was significantly higher in the 43°C group than those in the 32 and 38°C groups (F = 12.776, P < 0.001). Moreover, at T4, the serum levels of TGF-β (F = 3286.548, P < 0.001) and IL-10 (F = 4628.983, P < 0.001) were significantly increased in the 38°C group, and the serum levels of TNF-α (F = 911.415, P < 0.001) and IL-1β (F = 322.191, P < 0.001) were significantly decreased in the 38°C group, compared with the levels in the 32 and 43°C groups. CONCLUSION: Force-air warming blankets set at 38°C maintained stable body temperature with less adverse outcome and effectively inhibited the inflammatory response in pediatric patients undergoing surgery for DDH. CLINICAL TRIAL REGISTRATION ChiCTR1800014820; http://www.chictr.org.cn/showproj.aspx?proj=25240.