AIM To study the amino acids and proteins in 16 batches of commercial fish swim-bladders with different origins.METHODS A high performance liquid chromatography method based on pre-column derivatization using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate(AQC)was developed for the determination of contents and components of 17 amino acids in fish swim-bladders.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)was performed to analyze the molecular weight distribution of proteins from different fish swim-bladders,and proteins in fish swim-bladders were identified by proteomics method.RESULTS The result showed that the determination of 17 amino acids had a good linear relationship(R2≥0.998 0).The average recovery rate was 85.62%-109.60%and the relative standard deviations of precision,stability and repeatability were less than 3.5%.The total content of the 17 amino acids in 16 batches of fish swim-bladders ranged from 468.31 mg/g to 620.05 mg/g.A total of 688 proteins including 11 collagens were identified from 16 batches of fish swim-bladder samples and a plenty of low-abundance proteins at 52-95 kDa were also detected in fish swim-bladders by SDS-PAGE.CONCLUSION This study provides a good reference for the quality evaluation and further utilization of fish swim-bladders.

Seven compounds were isolated from fermentation extract of cave-derived Metarhizium anisopliae NHC-M3-2 by silica gel, semi-preparative HPLC and other chromatographic methods. Their structures were elucidated by UV, IR, MS and NMR methods as 2,3-dehydroindigotide G (1), (-)-regiolone (2), naphtho-γ-pyrone (3), indigotide G (4), indigotide B (5) destruxin A (6) and destruxin B (7). Compound 1 is a new glycoside naphthopyranone compound. The anti-hepatitis B virus (HBV) activity of these compounds was evaluated. The EC₅₀ and CC₅₀ of compound 3 against HBV were 4.5 μmol·L^-1 and 92.3 μmol·L^-1, respectively. This is the first report of the antiviral activity of compound 3.

The study aims to explore the intervention mechanism of Tingli Dazao Xiefei Decoction on asthma from the perspective of immune inflammation and intestinal flora, providing a theoretical basis for guiding clinical medication. The ovalbumin (OVA) asthmatic rat model was established by intraperitoneal injection of OVA sensitization solution and aerosol challenge, and divided into control (CON), model (M), dexamethasone group (DEX, 0.075 mg·kg^-1) and Tingli Dazao Xiefei Decoction (TLDZ, 3.5 g·kg^-1). Firstly, the effects of Tingli Dazao Xiefei Decoction on asthma symptoms of rats, lung and trachea pathological changes of asthmatic rats were observed by inducing cough and asthma experiment, phenol red excretion, hematoxylin-eosin staining (H&E), Masson and periodic acid Schiff (PAS) staining; the levels of transforming growth factor β1 (TGF-β1), interleukin (IL) 6 and IL-10 in rat serum and the levels of interferon γ (IFN-γ), immunoglobulin E (IgE), IL-4, IL-17A and tumor necrosis factor α (TNF-α) in bronchoalveolar lavage fluid (BALF) were detected by ELISA; the mRNA levels of IL-5, IL-13 and IL-33 in the lung were determined by qRT-PCR; the levels of macrophages and neutrophils in the spleen and the levels of natural killer cell (NK), helper T cell (Thc), dendritic cell (DC), regulatory T cell (Treg) and T helper cell 17 (Th17) in the peripheral blood were measured by flow cytometry combined with immunohistochemistry; the intestinal flora of asthmatic rats were analyzed by 16S rDNA high-throughput sequencing. Pathology and inflammatory results showed that Tingli Dazao Xiefei Decoction could effectively alleviate the asthma symptoms in rats, improve the pathological changes of lung tissue, reduce the production of goblet cells and collagen fibers, and reduce the inflammatory response in asthmatic rats; the results of immune cells showed that Tingli Dazao Xiefei Decoction could effectively increase the levels of NK, Thc, DC and Treg cells and reduce the levels of macrophages, neutrophils and Th17 cells in asthmatic rats; the results of intestinal flora showed that Tingli Dazao Xiefei Decoction could increase the levels of Lactobacillus, Ruminococcus, Christensenellaceae, Bifidobacterium and Eubacterium]_xylanophilum-group, and decrease the levels of Firmicutes, Desulfovibrio, Mucispirillum and Romboutsia in asthmatic rats. Therefore, it is speculated that Tingli Dazao Xiefei Decoction can improve the symptom of asthmatic rats by regulating the immune inflammation and intestinal flora in the asthmatic rats. All animal experiments in this article were approved by the Ethics Committee of Henan University of Chinese Medicine.

【Objective】 To explore the effects and the possible mechanism of RNA targeting membrane-bound prostaglandin E2 synthase l（mPGES- 1）on proliferation，apoptosis and drug resistance of leukemia cell line K562/A.【Methods】RNA interference was used to inhibit the expression of mPGES-1 of K562/A cells. Four groups were set up as follows：untreated group（K562/A），negative control group after interference（K562/A-NC），group after interference（K562/ A-KD），and group after interference with exogenous PGE2（K562/A-KD+PGE2）.Cell viability was assessed by CCK-8 assay. Cell apoptosis was analyzed by flow cytometry. Concentration of PGE2 was detected by ELISA. Proteins expression was detected by western blot.【Results】The expression of mPGES- 1 in K562/A cells was significantly down- regulated and the synthesis of PGE2 decreased（P < 0.000 1）after RNA interference. After RNA interference，the proliferation of K562/A cells was inhibited and apoptosis increased，and the sensitivity to chemotherapy drugs was enhanced（P < 0.05）. Meanwhile，the expression of β-catenin and MDR1 was decreased（P < 0.01）. Exogenous PGE2 could reverse the effect of RNA interference on proliferation ，apoptosis and drug sensitivity in K562/A cells（P < 0.05），and up-regulate the expression of β-catenin and MDR1（P < 0.01）. XAV939，an inhibitor of β-catenin，could down-regulate the expression of β- catenin and MDR1 in an dose- dependent pattern in K562/A cells（P < 0.05）.【Conclusions】RNA interference of mPGES- 1 could inhibit proliferation，induce apoptosis and reverse drug resistance in K562/A cells. The mechanism was related to reducing the synthesis of PGE2 and thus down- regulating the expression of β- catenin and MDR1. Wnt/β- catenin signal pathway may participate in the regulation of MDR1 by mPGES-1/PGE2.

Objective To understand the current situation of soil-transmitted nematode infections and influencing factors of pupils in rural areas of Yi Ethnicity in Butuo County, Sichuan Province, so as to provide the evidence for the prevention and control of soil-transmitted nematode infections. Methods In 2016, the pupils of 2 villages in Butuo County were selected by the multi-stage random sampling method, and the relevant information was obtained by questionnaires, including social demographic information, diet and hygiene habits and acknowledge of parasitic disease prevention. The soil-transmitted nematode infections were examined by Kato-Katz technique. The results were statistically analyzed by the single factor analysis and multivariate logistic regression methods. Results A total of 160 pupils were investigated in this survey. The soil-transmitted nematodes included Ascaris lumbricoides, Trichuris trichiura and hookworm. The overall infection rate was 89.38%, and the multi-infection rate was 54.55%. The infection rates of A. lumbricoides and T. trichiura were higher. The infection densities were mild (75.68%) or moderate (22.97%). The logistic regression analysis showed that the protective factor of soil-transmitted nematode infections was the knowledge about correct diet and health habits notified by teachers (OR = 0.67), but the risk factor was drinking unboiled water (OR = 19.26). Conclusions The infection rate of nematodes is still high in the pupils in rural areas of Yi Ethnicity in Butuo County, Sichuan Province. Therefore, we should strengthen the prevention and control of soil-transmitted nematode infections, especially A. lumbricoides and T. trichiura infections, and mostly develop the teachers’function in health education.

OBJECTIVE: To investigate the alternative antimicrobial drugs for the treatment of neonatal pertussis and the antigen genotypes of Bordetella pertussis (B. pertussis) strains. METHODS: A total of 32 B. pertussis strains isolated from neonates between May 2013 and July 2018 were used in this study. E-test stripes were used to measure the minimal inhibitory concentration (MIC) of 18 antimicrobial drugs including erythromycin, sulfamethoxazole-trimethoprim (SMZ) and ampicillin. The 23S rRNA gene of isolated strains was amplified and sequenced to identify the mutation site of erythromycin resistance gene, and the seven antigen genotypes of B. pertussis strains (ptxA, ptxC, ptxP, prn, fim2, fim3 and tcfA2) were analyzed. RESULTS: Of the 32 B. pertussis strains, 25 (78%) were resistant to erythromycin, azithromycin, clarithromycin and clindamycin, with an MIC of >256 mg/L, and A2047G mutation was observed in the 23S rRNA gene. All strains had an MIC of ≤0.064 mg/L for SMZ. The MIC of ampicillin, amoxicillin, amoxicillin-clavulanic acid and ceftriaxone ranged from 0.032 to 1 mg/L. The strains resistant to macrolide antibiotics had an antigen genotype of ptxA1/ptxC1/ptxP1/prn1/fim2-1/fim3-1/tcfA2. CONCLUSIONS B. pertussis strains from neonates are often resistant to macrolides, and the in vitro test shows that off-label use of sulfonamides is a reliable regimen for the treatment of neonates with macrolide-resistant pertussis. The prevalence of drug-resistant strains further emphasizes the importance of immunoprophylaxis.
Anti-Bacterial Agents ; Bordetella pertussis ; genetics ; Erythromycin ; Genotype ; Humans ; Infant, Newborn ; Microbial Sensitivity Tests ; Whooping Cough

Objective To investigate the distribution of alleles in 19 autosomal short tandem repeat (STR) loci in Jiangsu Han population.Methods Goldeneye20A kit was used to detect 9 025 samples.Genetic analysis was performed on typing data of 19 autosomal STR loci, and genetic distance with other 17 populations was analyzed.Results All the 19 autosomal STR loci were consistent with the Hardy-Weinberg equilibrium (P＞0.05), with the heterozygosity 0.616 1-0.916 3, probability of match 0.012 8-0.202 6, discrimination power 0.797 4-0.987 2, probability of paternity exclusion 0.310 8-0.828 8, and polymorphic information content 0.561 7-0.913 6.The cumulative discrimination power and cumulative probability of exclusion were 0.999 999 999 999 999 998 434 1 and 0.999 999 989, respectively.The Jiangsu Han population had close genetic distances with the Han population in Tianjin, Hunan and Jilin, and significant difference with Han population in Aletai region in Xinjiang (P＜0.05).Conclusion The STR allele polymorphism data and population genetic parameters of Jiangsu Han population can provide data support for the forensic application of these STR loci in Jiangsu Han population.

Objective Based on the disadvantages of NUSS procedure, we design a new type of funnel chest orthodontic steel bars for clinical application to simplify surgery operation , reduce the surgical trauma, improve operation effect, enhance the safety of operation, and relieve the suffering of the patients, etc.Methods From June 2010 to July 2015, 1020 patients used the new type of funnel chest orthodontic metal plates, including 706 cases of males and 314 cases of females, aged 3-40(aver-age age 11.38 ±5.31).Haller index of 3.36 to 15.45(average index 4.97 ±1.58).668 symmetrical funnel chest cases, 281 cases of asymmetry and 71 recurrent funnel chest cases are involved.Surgical steel bars, having 15 specifications, are arc-shaped, the lengths of which are from 12cm to 26cm.The two ends of steel bars have fixed piece, one end of the bar was fused with a stabilizer and the other end connected to an introducer or a stabilizer and use screws to fix two ends .Steel plates, as the design product, are finalized in the factory.Therefore we need not make intraoperative reprocess.Front tip of introducer and the back-end can be connected to the metal plate.Patients were placed supine position under general anesthesia.Two inci-sions, with length about 2cm and deep to the rib periosteum were made along axillary midline line on the both sides .In the di-rection of high blunt slightly medial free for subcutaneous tunnel to the highest point .Fixed wires were worn around the ribs on both sides.A thoracoscope to guide the bar was inserted from the highest point on the right side of the rear of the medial by sternum low into the chest from the left corresponds to the highest point of the medial , remove the introducer to connect another stator, use screws to fix the connection, with a fixed stator and steel plate stabilized.At the end of the surgery, the stabilizer on either side was secured with sutures.Results The median first operation time was 23 -45(32.31 ±3.57)min, second op-eration time was about 34-147(68.27 ±30.25)min..The bleeding volume(first operation) was 0-2(0.40 ±0.36)ml, the bleeding of second operation time was 1-630(144.00 ±57.00)ml.The hospital stay was 3-9(4.22 ±0.61)days after the operation.All the cases were followed up for 34-48(34.4 ±6.1)months.Steel plate was fixed at the bottom of the sternum in 957 of them,with slight shifting in 58 patients.There was another 5 cases who need re-operation because of the obvious shift-ing.Conclusion The new type of funnel chest orthodontic steel bars simplify surgery operation, reduce the surgical trauma, improve operation effect, relieve the suffering of the patients and enhance the safety of operation.

The technique of liquisolid compress is a new technique developed in 1990s, which was considered to be the most promising technique to improve the dissolution of water-insoluble drugs. In this article, tanshinone II(A) and the extracts of the ester-solubility fractions were chosen as the model drugs to evaluate the effects of the liquisolid technique for enhancement of dissolution properties of tanshinone II(A). Several liquisolid tablets (LS) formulations containing different dosage of drugs and various liquid vehicle were pre-pared and for all the formulations, microcrystalline cellulose and silica were chosen as the carrier and coating materials to evaluate their flow properties, such as angle of repose, Carr's compressibility index and Hausner's ratio. The interaction between drug and excipients in prepared LS compacts were studied by differential scanning calorimetry(DSC) and X-ray powder diffraction (XRPD). The dissolution curves of tanshinone II(A) from liquisolid compacts were investigated to determine the technique's effect in improving the dissolution of tanshinone II(A) and its impacting factors. According to the results, the dissolution increased with the rise in the dissolution of the liquid-phase solvent. The R-value and drug dosage can significantly affect the drug release, but with less impact on active fractions. This indicated that liquisolid technique is a promising alternative for improvement of dissolution property of water-soluble drugs, and can make a synergistic effect with other ester-soluble constituents and bettern improve the release of tanshinone II(A). Therefore, the technique of liquisolid compress will have a better development prospect in traditional Chinese medicines.
Calorimetry, Differential Scanning ; Diterpenes, Abietane ; chemistry ; Solubility ; X-Ray Diffraction

To develop a HPLC method for determination of the concentration of scutellarin and scutellarin ethyl ester and their pharmacokinetics were also compared. 104 mg kg-1of scutellarin or 114. 5 mg kg-1 scutellarin ethyl ester were given at single dose by oral gavarge. Blood samples were collected from the jugular vein. Plasma concentration was measured by HPLC. The pharmacokinetic parameters were calculated with Winnonlin program. The plasma concentration-time profile of scutellarin and scutellarin ethyl ester were both fitted with non-compartment model and both were double peaks. The main pharmacokinetic parameters of scutellarin and scutellarin ethyl ester were as follows: Tmax Cmax and AUC0-t for scutellarin were (6 +/- 1.26) h, (321.55 +/-48.31) microg L-1 and (2 974 +/-753) h micro.g L-1; for scutellarin ethyl ester, Tmax, Cmax and AUC0-t were 0.5 h, (1 550.82 +/-219.75) +/- microg L- and (6 407 +/- 399) h microg L-1. The speed ingested into the blood of scutellarin ethyl ester was faster than scutellarin, and the bioavailability of scutellarin ethyl ester was two times higher than scutellarin.
Animals ; Apigenin ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Flavones ; pharmacokinetics ; Glucuronates ; pharmacokinetics ; Glucuronides ; pharmacokinetics ; Male ; Rats ; Rats, Wistar