AIM: To detect the effects of Xinmailing Solution and MK-801 on injury of neuronal cell induced by glutamate. METHODS: Cultured neuronal cell injuried by glutamate was prepared and the content of malondialdehyde and nitrite in cell supernatant was measured. Morphology changes were also observed with discrepancy microscope at the same time. RESULTS: Xinmailing Solution and MK-801 attenuated cell injury induced by glutamate,and inhibited increase in malondialdehyde and nitrite in cell supernatant. CONCLUSION: Xinmailing Solution had a protective effect on neuronal cell at cell level.

AIM: To establish the quality standard for Lixieling Pill(Herba Andrographis; Radix Sophorae flavescentis etc.). METHODS: Herba Andrographis and Radix Sophorae flavescentis in Lixieling Pill were identified by TLC. Andrographolide, dehydroandrographolide, matrine and oxgmatrine of pill were determined by HPLC together. RESULTS: The characteristic identification by TLC was distinct and highly specific. The quantitative evaluation of andrographolide had the linear range of 0.969 -2.422 ?g, The average recovery was 98.45% and RSD was 0.88. The quantitative evaluation of dehydroandrographolide had the linear range of 0.481 -1.204 ?g , The average recovery was 100.53% and RSD was 0.63. The quantitative evaluation of matrine had the linear range of 1.032 -2.064 ?g , The average recovery was 99.06% and RSD was 1.04. The quantitative evaluation of oxgmatrine had the linear range of 1.016 -2.032 ?g , The average recovery was 99.67% and RSD was 1.20. CONCLUSION: The method is reliable, accurate and specific. It can be used for quality control of Lixieling Pill.

Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Objective To establish the theoretical basis of the probiotic application among very low birth weight(VLBW) infants,the efficacy of probiotics on the gut microbiota of VLBW infants on the 14th postnatal day was studied.Method The VLBW infants admitted to BaoAn Maternal and Child Care Hospital from January 2015 to December 2015 were randomly assigned into probiotics group and placebo group.From the first feeding to corrected gestational age of 36 weeks,probiotics group was treated with a combination of Bifidobacterium and Lactobacillus while placebo group with placebo.Fecal samples were collected on the 1st and 14th postnatal day.Total bacterial DNA was extracted and sequenced using high-throughput 16S rRNA gene sequencing on MiSeq sequencing platform.Result A total of 21 VLBW infants were enrolled,9 in probiotics group and other 12 placebo group.No significant differences of clinical data existed between the two group (P ＞ 0.05),The abundance and diversity of microflora (P ＞ 0.05) on the first day between the two group were similar.Compared with placebo group,the relative abundance of Bifidobacterium and Lactobacillales in stool samples on the 14th day was significantly increased,while the Halomonas was significantly decreased.The relative abundance of the Shannon-index was increased,but without significant difference (P =0.16).Conclusion Enteral supplementation of probiotics in VLBW infants may increase probiotic bacterium and microflora diversity.

Objective To investigate the relationship between serum homocysteine ( Hcy ) and cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD) after stroke, and observe the plasma Hcy levels and cognitive function improvement when treated with folic acid and vitamin B12 .Methods 87 acute cerebral infarction combined with COPD patients as the research object, then the general clinical data were recorded, hematology indexes ( Hcy, folic acid, vitamin B12 ) were determined, and their cognitive function with a simple mental state scale (MMSE) was assayed.According to the plasma Hcy levels, the subjects were divided into Hcy-normal group (n =21) and Hcy-increased group (n=66), then compare the cognitive function between the two groups.Hcy-increased subjects were randomly divided into intervention group (conventional treatment +folic acid 2.5 mg +VitB12 0.5 mg) and control group (conventional treatment).After six months follow-up, we retested plasma Hcy levels and MMSE assessment, comparison of plasma Hcy concentration change and cognitive function improvement between two groups.Results Compared with Hcy-normal group, plasma folic acid, VitB12 levels significantly decreased in Hcy-increased group (P<0.05).And Hcy concentration was negatively correlated with folic acid(r=-0.351,P =0.000)and VitB12(r=-0.242,P=0.015)levels.In addition, the MMSE, directional force and delayed recall score decreased in Hcy-increased group compared with the Hcy-normal group ( P<0.05 ).Hcy levels were significantly lower than the baseline level (P<0.05), MMSE and the sub-project of cognitive function score increased after treated with folic acid and VitB12 for six months, although there was no statistically significant difference.Conclusion Plasma Hcy level is associated with cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD), patients treated with folic acid and VitB12 may help slow the recent cognitive dysfunction after stroke in the near future.

Objective To investigate the carrying status and characteristics of Clostridium difficile isolated from infants.Methods Two hundred and thirty-eight stool specimens were collected from infant younger than 1 year old,that were hospitalized or outpatient from August to November 2015.Immunochromatography targeted GDH and toxin A&B of C.difficile was used for C.difficile screening,and those positive specimens were inoculated in CDIF and anaerobic culture.C.difficile isolates were genotyped by using slpA sequence typing (slpA ST),and tcdA,tcdB,cdtA and cdtB of C.difficile isolates were detected by PCR.Results Fifty C.difficile strains were isolated from 238 stool samples,and the isolated rates of C.difficile from <3 months,3 months to <6 months,and 6 months to 1 years old groups were 9.3%,17.6% and 27.3%(χ2=6.940,P=0.031<0.05),respectively.52.0%(26/50) of the C.difficile isolates were toxigenic,and 69.2% (18/26) toxigenic isolates harbored tcdA+tcdB+cdtA-cdtB-.Fifty C.difficile isolates were genotyped as 11 slpA STs,slpA ST fr-02 and kr-02 were the commonest genotypes in toxigenic C.difficile isolates;however,that was slpA ST xr-03 in non-toxigenic isolates.Conclusion High C.difficile carriage is found in infants younger than 1 year old,and more than half of C.difficile isolates are toxigenic.Most of toxigenic isolates harbored toxin A and B.The genotype of C.difficile isolates is different between toxigenic isolates and non-toxigenic isolates.

Objective:To study the feasibility of preserving the subvalvular structure during mitral valve replacement and whether it is more conducive to the recovery of cardiac function.Methods:The clinical data of 205 patients who underwent mitral valve replacement due to mitral insufficiency in Enshi Central Hospital from January 2017 to December 2020 were analyzed retrospectively. According to whether the subvalvular structure of mitral valve was preserved during operation, they were divided into three groups: group A was to preserve all the subvalvular structures of anterior and posterior mitral valve, group B retained all or part of the subvalvular structure of the posterior valve, in group C, all anterior and posterior subvalvular structures were removed. The operation time, cardiopulmonary bypass time and aortic occlusion time of patients in each group were counted. Postoperative ventilator use time, ICU stay time and incidence of complications; mitral flow velocities (MVE), left ventricular end diastolic dimension (LVED), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were rechecked by echocardiography at 1 week, 3 months and 6 months after operation.Results:There were no significant differences in operation time, cardiopulmonary bypass time, aortic occlusion time, ventilator use time and ICU stay time among the three groups before surgery ( P>0.05). There were no death cases in perioperative period, and all patients were cured and discharged from hospital. The results of cardiac color ultrasound showed that MVE, LVED, LVEF and LVFS were improved compared with those before operation. Postoperative ultrasound results showed no significant difference in MVE at 1 week, 3 months and 6 months ( P>0.05). At 1 week after surgery, LVED in group A was lower than that in group B and C: (44.90 ± 5.59) mm vs. (46.13 ± 6.15) mm, (48.33 ± 5.59) mm. The difference was statistically significant ( P<0.05). Three months after operation, LVED and LVFS in group A were higher than those in group B and group C: (43.37 ± 5.61) mm vs. (44.69 ± 5.45) mm and (53.42 ± 5.35) mm, (33.92 ± 3.34)% vs. (31.67 ± 3.20)% and (30.37 ± 6.96)%, respectively. The differences were statistically significant ( P<0.05). At 6 months after operation, LVED in group A was lower than that in group B and group C: (41.86 ± 3.27) mm vs. (47.85 ± 6.93) mm and (53.42 ± 8.17) mm. LVFS was higher than that in group B and group C: (33.42 ± 2.64) % vs. (32.18 ± 5.98) % and (32.28 ± 2.58) %, and the differences were statistically significant ( P<0.05). Conclusions:Preserving the whole subvalvular structure of mitral valve will not lead to prosthetic valve dysfunction and increase surgical complications. After operation, it showed better reduction of left ventricular end diastolic diameter and better left ventricular shortening rate, which was more conducive to the recovery of cardiac function.

Objective:To discuss the improvement effect of curcumin combined with fecal bacteria transplantation on the mice with dextran sulfate sodium(DSS)-induced ulcerative colitis(UC),and to clarify the related mechanism.Methods:Fifty mice were randomly divided into control,model,curcumin,fecal bacteria transplantation,and combination groups.Except for the mice in control group(given distilled water),the mice in the other groups were given distilled water containing 2%DSS to establish the UC models.The mice in curcumin group were gavaged with 0.4 mL of 60 mg·kg-1 curcumin solution once per day for 10 d;the mice in fecal bacteria transplantation group underwent enema with 0.2 mL of fecal bacteria suspension once per day for 10 d;the mice in combination group received the enema of 0.2 mL fecal bacteria suspension and gavaged with 0.4 mL of 60 mg·kg-1 curcumin solution.At the end of the experiment,the disease activity index(DAI)and colon macroscopic damage index(CDMI)of the mice in various groups were calculated;the morphology of colon tissue of the mice in various groups was detected by HE staining;the levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-4,and IL-10 in colon tissue of the mice in various groups were detected by enzyme-linked immunosorbent assay(ELISA)method;the expression levels of occludin and zonula occludens-1(ZO-1)mRNA and proteins in colon tissue of the mice in various groups were detected by real-time fluorescence quantitative(RT-qPCR)and Western blotting methods.Results:The intestinal mucosal epithelial structure of the mice in control group was intact and continuous with regular glandular arrangement and without inflammatory cell infiltration or ulceration;the intestinal mucosal epithelial structure of the mice in model group exhibited loss of colonic mucosal epithelium,disordered glandular arrangement,reduced goblet cells,congestion and edema in mucosal and submucosal layers,and extensive infiltration of inflammatory cells with widespread small ulcers;the intestinal mucosal epithelial structure of the mice in curcumin,fecal bacteria transplantation,and combination groups exhibited relatively intact colonic mucosal epithelial structures with reduced inflammatory cell infiltration and ameliorated mucosal and submucosal congestion and edema.Compared with control group,the DAI and CDMI of the mice in model group were increased(P<0.05),the levels of IL-1β and TNF-α were increased(P<0.05),the levels of IL-4 and IL-10 were decreased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were decreased(P<0.05);compared with model group,the DAI and CDMI of the mice in curcumin,fecal bacteria transplantation,and combination groups were decreased(P<0.05),the levels of IL-1β and TNF-α were decreased(P<0.05),the levels of IL-4 and IL-10 were increased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were increased(P<0.05).Compared with curcumin group and fecal bacteria transplantation group,the DAI and CDMI of the mice in combination group were decreased(P<0.05),the levels of IL-1β and TNF-α were decreased(P<0.05),the levels of IL-4 and IL-10 were increased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were increased(P<0.05).Conclusion:Curcumin combined with fecal bacteria transplantation can ameliorate the pathological damage in colonic tissue of the UC mice,inhibit the secretion of inflammatory factors,and promote the repaiment of intestin mucosa.