Objective To investigate the feasibility of the gene chip technique and multiplex PCR technique for detecting and screening foodborne pathogens .Methods The primer sequences were designed to target cells ,the biotin was adopted to label the re‐verse primer 5′end and the amino group was adopted to label oligonucleotide probe 5′end .The probe spotted on a solid support for preparing microarray ,PCR product was hybridized with microarray probe region ,PCR and hybridization reaction system was opti‐mized .Results The microarray technique could simultaneously detect multiple pathogens of Shigella ,Salmonella ,Klebsiella pneu‐moniae ,Brucella ,Proteus mirabilis ,Staphylococcus aureus ,Campylobacter jejuni ,etc .,which was easy to operate and had strong specificity .The sensitivity of bacterial pure cultures was 5 .0 × 102 CFU/mL ,the sensitivity of DNA detection was 0 .1 pg ,the coin‐cidence rate for detecting isolated bacteria was 100 % .The PCR detection system was established and optimized by using primers , the concentration of Mg2+ and the annealing temperature Tm value of 1 .5 mmol/L and 56 ℃ were determined ,the detection sensi‐tivity reached to 10 pg ,all the specific primers amplified bands could be amplified under this sensitivity .Conclusion The gene chip technique and multiplex PCR technique can effectively detect foodborne pathogens ,which provide a new idea for detecting pathogens with the high‐throughput screening and are worth popularization and application in the field of food safety .

Objective To study the distribution of clinically separated Hemophilus (H .) influenzae and its drug resistance situa‐tion .Methods Among 2 663 specimens of screening qualified and positive culture collected from the outpatients ,inpatients and e‐mergency patients of our hospital from January to December 2014 ,including 1 895 specimens of lower respiratory tract ,666 blood specimens and 102 urinary secretion specimens ,438 strains of H .influenzae were isolated .The cephalosporin nitro thiophene paper test strip was used to test its beta lactamase and the K‐B method was used to detect its sensitivity to 8 kinds of antibacterial drugs . The testing data were statistically analyzed by using WHONET 5 .6 software .Results Among 438 strains of H .influenzae ,98 .6%was originated from hospitalized patients ,1 .4% from the outpatient department ;176 isolated strains were originated from children , accounting for 40 .18% ;194 strains (44 .29% ) were from elderly patients ,434 strains(99 .09% ) were originated from the lower re‐spiratory tract specimens .The resistance rate to compound sulfamethoxazole was highest and reached 82 .90% ,which to ampicillin and tetracycline were 29 .00% and 43 .40% respectively ;but which to cefuroxime ,ceftriaxone ,ciprofloxacin ,azithromycin ,imipenem were below 2 .00% .The β‐lactamase positive rate was 43 .40% ,in ampicillin resistant strains ,1 .57% of strain was negative in β‐lac‐tamase test .The resistance to ampicillin and enzyme production rate had no significant differences between the children isolated strains and adult isolated strains (42 .43% ,42 .40% vs .44 .00% ,44 .00% ) .Conclusion H .influenzae is mainly isolated from low ‐er respiratory tract specimens ;which is dominated by children and elderly patients ;the second and third generations of cephalospo‐rins and azithromycin can serve as a clinical preferred drugs ;β‐lactamase is the primary mechanism of resistance to ampicillin .

Objective To analyze the effect evaluation of modified TT enrichment broth in the separation of intestinal pathogenic Salmonella and Shigella bacteria .Methods The routine inoculation of Macconkey agar and SS agar were adopted ,meanwhile two different methods for adding the modified TT enrichment broth(sodium thiosulfate and calcium carbonate) were used to screen Sal‐monella and Shigella bacteria .Serum coagulation was performed by referring to the bacterial isolation situation over the years .The SPSS 18 .0 software was adopted to process the data .Then the difference in the separation rate of Salmonella and Shigella bacteria between the two different stool culture methods was compared .Results Totally 790 stool culture samples during 2013-2015 were included into the statistical analysis ,30 cases of Salmonella bacteria were isolated by adopting the routine method ,the positive rate was 3 .80% ;5 cases were Shigella bacteria ,the positive rate was 0 .63% ;but 77 cases of Salmonella bacteria were isolated by adop‐ting the modified TT enrichment broth method ,the positive rate was 9 .75% ,7 cases were Shigella bacteria ,the positive rate was 0 .89% ;the detection rate of Salmonella bacteria by adopting the modified TT enrichment broth method was 2 .57 times of the con‐ventional culture method ,which of Shigella bacteria was 1 .41 times ,showing that the difference of Salmonella and Shigella bacteria isolation between the two different methods had statistical significance(P<0 .05) .Conclusion The modified TT enrichment broth can significantly improve the positive isolation rate of Salmonella and Shigella stool culture ,which provides larger help for clinical doctor′s correct diagnosis and treatment of patients .

To investigate the prevalence of mental health problems in male violent offenders of Hunan and Sichuan Provinces; and to compare the types and severity of problems between the violent and nonviolent offenders.Methods:Ninety-one violent juvenile offenders and 64 nonviolent juvenile offenders in the juvenile detention centers of Hunan,and 81 violent juvenile offenders in the juvenile detention centers of Sichuan; 39 high school students from a middle school of Hunan Province and 49 from a middle school of Sichuan Province were investigated using the Investigation Screening Inventory for Child Mental Disorder and the Kiddy Schedule for Affective Diseases and Schizophrenia (K-SADS-PL),and classified according to the Diagnostic and Statistical Manual of Mental Disorders.Results:In the Hunan violent group,86.6％ met the criterion of conduct disorder (CD),17.6％ of substance abuse,9.9％ of substance dependence,while in the Hunan nonviolent group,75.0％ met the criterion of CD,and 11.7％ of substance abuse; these were all significantly higher than those of the control group (P＜0.05).In the Sichuan violent group,17.3％ met the criterion of attention deficit/hyperactivity disorder (ADHD),18.5％ of oppositional defiant disorder (ODD),69.1％of CD,and 22.2％ of substance abuse; these were all significantly higher than those of the Sichuan control group.Moreover,63.7％ of Hunan-violent,55.6％ of Sichuan-violent,and 45.0％ of Hunannonviolent offenders had a previous offence records.Conclusion:Mental and behavioral disorders among delinquent youth is becoming a serious problem,and there is an urgent need to develop and implement effective assessment and treatment approaches for juvenile offenders with the aim of reducing offence and recidivism in this population.

OBJECTIVE To screen the anti-meticillin-resistant Staphylococcus aureus(MRSA)traditional Chinese materia medica(TCMM)by biosensor technique,targeted on the soluble penicillin binding protein 2a(PBP2a)of clinical MRSA.METHODS The soluble PBP2a with amino acid sequence from 25 to 668 from clinical MRSA were expressed in Escherichia coli by gene recombination technique.Then,the expressed product was identified and its biological function was analyzed.After the PBP2a was immobilized into the carboxymethyl dextran cuvette(CMD),the anti-MRSA TCMM was screened by means of biosensor.RESULTS The soluble protein PBP2a had been successfully expressed,whose relative molecular mass was 74?103.It was confirmed that the soluble PBP2a had transpeptidase activitiy and ?-lactamase activitiy.Subsequently,10 kinds of anti-MRSA TCMM were screened out by biosensor technique.Moreover,Radix Scutellariae,Rhizoma Coptidis and Spica Prunellae had greater anti-MRSA effect than others.CONCLUSIONS Anti-MRSA TCMM has been successfully screened out by biosensor technique,targeted on the soluble PBP2a of clinical MRSA.

Objective Research the bilingual teaching of medical organic chemistry experiment. Methods PBL teaching method was used in bilingual teaching of medical organic chemistry experiment. Results As a result, we improved the effect of bilingual teaching in medical organic chemistry experiment. Conclusion Teaching research found that the teaching mode with a problem-centered and students-orientation approach has improved students' analysis and problem solving skills. During the process of teaching, the students have enhanced their enthusiasm of study.

OBJECTIVE:To study the mechanism of Aurantii fructus immaturus(AFI)and its main active ingredients in pro-moting gastrointestinal motility of model rats with spleen deficiency. METHODS:170 rats were randomly divided into blank group (10 rats) and modeling group (160 rats),rats in modeling group was induced models with spleen deficiency by bitter cold diar-rhea+irregular diet. After modeling, rats were randomly divided into model group, naringin (NA) low-dose, medium-lose, high-dose groups(3.267,6.535,13.070 mg/mL),neohesperidin(NE)low-dose,medium-lose,high-dose groups(3.865,7.730, 15.460 mg/mL),synephrine(SY)low-dose,medium-lose,high-dose groups(0.252,0.504,1.008 mg/mL),compatibility groups with 3 monomer ingredients (NA-NE-SY) low-dose,medium-lose,high-dose and AFI water decoction low-dose,medium-lose, high-dose groups(0.104,0.208,0.416 g/mL,calculated by crude drug),ig,once a day,10 mL/kg,for 7 d. After the last admin-istration,gastrin (GAS) in serum,and acetylcholine (ACh),motilin (MTL),substance P (SP),vasoactive intestinal peptide (VIP)levels in plasma were detected. RESULTS:Compared with blank group,GAS level in serum and ACh,MTL,SP levels in plasma in model group were reduced(P<0.01),VIP level in plasma was increased(P<0.05). Compared with model group,ex-cept for the GAS level in serum showed no obvious change in NA high-dose group and SY doses groups,other medicine groups were obviously increased (P<0.05 or P<0.01);the ACh levels in serum were obviouly increased in NE high-dose group,SY high-dose group and AFI water decoction low-dose group(P<0.01). MTL levels in plasma were obviously increased in NE medi-um-dose,high-dose groups,SY high-dose group,compatibility low-dose,medium-dose groups and AFI water decoction medi-um-dose,high-lose groups (P<0.05);SP levels in plasma were obviously increased in NA low-dose,medium-dose groups and NE doses groups(P<0.05 or P<0.01);VIP levels were reduced in NA low-dose group,SY high-dose group and AFI water decoc-tion low-dose,medium-lose groups(P<0.05). CONCLUSIONS:AFI may promote the gastrointestinal motility of model rats with spleen deficiency by promoting the secretion of GAS,ACh,MTL,and inhibiting the secretion of VIP;there are differences be-tween AFI and the 3 monomer ingredients in regulation of gastrointestinal hormones.

Objective To study the effect of Chinese medicinal compound recipe ( modified Fuzheng Yiliu Decoction, MFYD) on the proliferation of colon cancer HT-29 cells and on the expression of cellular apoptosis gene, Bcl-2, and tumor-inhibiting gene, P53. Methods Colon cancer HT-29 cells were divided into serum containing MFYD groups ( treated with different concentrations of serum containing MFYD) and blank serum group. Methyl thiazolyl tetrazolium ( MTT) , real time cellular analysis ( RTCA) and reverse transcription poly merase chain reaction ( RT-PCR) were used to detect the cell proliferation and mRNA expression of Bcl-2 and P53 separately. Results The proliferation of colon cancer HT-29 cell line was inhibited by MFYD ( P<0.05) in concentration-and time-dependent manner. The Bcl-2 mRNA expression was decreased and p53 mRNA was increased markedly in HT-29 cells after co-culturing with 12% or 15% volume fraction of serum containing MFYD for 48 hours, the differences being significant compared with the blank serum group ( P<0.05). Conclusion MFYD has obvious inhibitory effect on the proliferation of colon cancer HT-29 cells, and its mechanism may be related to down-regulating the expression of Bcl-2 gene and up-regulating the expression of P53 gene.

Objective:To explore the effect of electroencephalographic (EEG) biofeedback combined with buspirone on patients with schizophrenia with anxiety, in order to provide evidence-based reference for clinical treatment.Methods:From January 2018 to January 2020, 80 patients with schizophrenia and anxiety in Jinzhou Kangning Hospital were prospectively selected and divided into the study group and the control group by simple randomization, each group with 40 patients. The control group was given buspirone, and the study group was given EEG biofeedback combined with buspirone. Both were treated for 8 weeks. The efficacy, adverse reactions and the Positive and Negative Symptoms Scale (PANSS) and Hamilton Anxiety Scale (HAMA), the Pittsburgh Sleep Quality Scale (PSQI), Scale of Social Function in Psychosis Inpatients (SSPI), Barthel Index (BI) score, and serum neuroendocrine index thyroid stimulating hormone (TSH), cortisol (Cor) levels before treatment, 4 weeks and 8 weeks after treatment were compared between the two groups.Results:After 8 weeks of treatment, the total effective rate in the study group was higher than that in the control group: 95.0% (38/40) vs. 77.5% (31/40), the difference was statistically significant (χ 2 = 5.16, P<0.05). After 4, 8 weeks of treatment, the PANSS scores in the study group were lower than those in the control group: (49.57 ± 5.65) scores vs. (57.96 ± 6.48) scores, (37.69 ± 4.35) scores vs.(45.07 ± 5.74) scores, the differences were statistically significant ( P<0.05). After 4, 8 weeks of treatment, the HAMA and PSQI scores in the study group were lower than those in the control group, while the SSPI and BI scores were higher than those in the control group, the differences were statistically significant ( P<0.05). After 4, 8 weeks of treatment, the serum TSH and Cor levels in the study group were lower than those in the control group: after 4 weeks of treatment: (2.74 ± 0.84) mU/L vs. (3.35 ± 0.97) mU/L, (276.51 ± 45.96) μg/L vs. (346.42 ± 50.34) μg/L; after 8 weeks of treatment: (2.46 ± 0.72) mU/L vs. (2.82 ± 0.86) mU/L, (197.26 ± 36.84) μg/L vs. (264.19 ± 42.46) μg/L, the differences were statistically significant ( P<0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups during treatment ( P>0.05). Conclusions:EEG biofeedback combined with buspirone can enhance the therapeutic effect by regulating neuroendocrine, reduce the mental symptoms, anxiety and sleep disorders ofschizophrenia patients with anxiety. It can also improve social function and daily life ability, and have high safety.

Kv2.1 channel currents in pancreatic beta-cells are thought to contribute to action potential repolarization and thereby modulate insulin secretion. Because of its central role in this important physiological process, Kv2.1 channel is a promising target for the treatment of type 2 diabetes. Jingzhaotoxin-XI (JZTX-XI) is a novel peptide neurotoxin isolated from the venom of the spider Chilobrachys jingzhao. Two-microelectrode voltage clamp experiments had showed that the toxin inhibited Kv2.1 potassium currents expressed in Xenopus Laevis oocytes. In order to investigate the structure-function relationship of JZTX-XI, the natural toxin and a mutant of JZTX-XI in which Arg3 was replaced by Ala, were synthesized by solid-phase chemistry method with Fmoc-protected amino acids on the PS3 automated peptide synthesizer. Reverse-phase high performance liquid chromatography (RP-HPLC) and matrix assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were used to monitor the oxidative refolding process of synthetic linear peptides to find the optimal renaturation conditions of these toxins. The experiments also proved that the relative molecular masses of refolded peptides were in accordance with their theoretical molecular masses. RP-HPLC chromatogram of co-injected native and refolded JZTX-XI was a single peak. Under the whole-cell patch-clamp mode, JZTX-XI could completely inhibit hKv2.1 and hNav1.5 channels currents expressed in HEK293T cells with IC50 values of 95.8 nmol/L and 437.1 nmol/L respectively. The mutant R3A-JZTX-XI could also inhibit hKv2.1 and hNav1.5 channel currents expressed in HEK293T cells with IC50 values of 1.22 micromol/L and 1.96 micromol/L respectively. However, the prohibitive levels of R3A-JZTX-XI on hKv2.1 and hNav1.5 channels were reduced by about 12.7 times and 4.5 times respectively, indicating that Arg3 was a key amino acid residue relative to the hKv2.1 channel activity of JZTX-XI, but it is also an amino acid residue correlated with the binding activity of JZTX-XI to hNav1.5 channel. Our findings should be helpful to develop JZTX-XI into a molecular probe and drug candidate targeting to Kv2.1 potassium channel in the pancreas.
Animals ; HEK293 Cells ; Humans ; Insulin-Secreting Cells ; metabolism ; Mutant Proteins ; genetics ; pharmacology ; NAV1.5 Voltage-Gated Sodium Channel ; metabolism ; Neurotoxins ; chemical synthesis ; genetics ; pharmacology ; Protein Refolding ; Shab Potassium Channels ; antagonists & inhibitors ; metabolism ; Sodium Channel Blockers ; pharmacology ; Spider Venoms ; genetics ; pharmacology ; Transfection