This study was aimed at exploring the prevalence and drug sensitivity of Gordonia strains isolated from sputum samples in Henan Province,to provide data to aid in the prevention and treatment of Gordonia infection.A combination of 16S rDNA and sec A1 gene sequencing was used to identify the isolated strains,and susceptibility to16 drugs was determined with the broth microdilution method.A total of 21 strains were identified through 16S rDNA gene and sec A1 gene sequencing,including five strains of Gordonia broncians,eight strains of Gordonia paraphernivans,seven strains of Gordonia sputi,and one strain of Gordonia aichiensis.Drug sensi-tivity testing showed high Gordonia sensitivity to drugs such as ceftriaxone,linezolid,doxycycline,amoxicillin/clavulanic acid,mino-cycline,cefotaxime,trimethoprim/sulfamethoxazole,imipenem,tobramycin,and clarithromycin.The sensitivity rates of the isolated strains were 90.48%(19/21),100%(21/21),90.48%(19/21),90.48%(19/21),95.24%(20/21),90.48%(19/21),90.48%(19/21),90.48%(19/21),and 95.24%(20/21),respectively.Gordonia showed high resistance to rifampicin and cefepime,with rates of 28.57%(6/21)and 19.05%(4/21),respectively.Meanwhile,the resistance varied among bacterial strains.The resistance rate of G.sputi to rifampicin reached 71.43%(5/7),whereas that of G.parapffinivoras to cefepime was 37.5%(3/8).The main species of Gordo-nia isolated from sputum samples of patients in Henan Province were G.bronchialis,G.paraffinivoras,G.sputi,and G.aichiensis.Drug sensitivity tests indicated that drugs including amoxicillin/clavulanic acid,ceftriaxone,cefotaxime,tobramycin,clarithromycin,mi-nocycline,trimethoprim/sulfamethoxazole,linezolid,and doxycycline had good antibacterial effects against Gordonia.

The rabies virus(RABV)that causes rabies mainly attacks the peripheral and central nervous systems.In the later stages of infection,it is scattered in the salivary glands and transmit-ted to other susceptible animals through infectious saliva.To study dispersion of the RABV in the three pairs of salivary gland tissues,the street strain PB4 of the RABV was inoculated into 21-day-old female mice through the hind limb muscles.During the moribund stage of the mice,the sublin-gual gland,submandibular gland and parotid gland were collected,respectively.The TCID50 titer of RABV in the three kinds of glands of the mice and the copy number of the RABV N gene were de-tected,and RABV in different salivary glands was observed by immunofluorescence.The results showed that PB4 was dispersed in all three kinds of salivary glands of the mice,with the largest a-mounts in the parotid gland,followed by the submandibular gland,and the lowest amount in the sublingual gland.Three-month-old dogs were inoculated with PB4 through the cranial cavity,and saliva were collected every 12 h after inoculation.The saliva samples were detected by TCID50 and RT-qPCR.And during the moribund stage of the dogs when the disease occurred,the three pairs of salivary glands were collected.Through the determination of the TCID50 titer,RT-qPCR and immu-nofluorescence detection,it was demonstrated that among the three different salivary glands of the dogs,the largest amount of PB4 was found in the parotid gland and the lowest in the sublingual gland.Our results in mice and dogs clearly proved that the parotid gland was consistently found to exhibit the highest content of street RABV among the three major salivary glands,which could en-rich experimental data for analyzing the dispersion of RABV in the salivary glands and interpreta-tion of the intermittent secretion of saliva in clinically rabid dogs.

The rabies virus(RABV)that causes rabies mainly attacks the peripheral and central nervous systems.In the later stages of infection,it is scattered in the salivary glands and transmit-ted to other susceptible animals through infectious saliva.To study dispersion of the RABV in the three pairs of salivary gland tissues,the street strain PB4 of the RABV was inoculated into 21-day-old female mice through the hind limb muscles.During the moribund stage of the mice,the sublin-gual gland,submandibular gland and parotid gland were collected,respectively.The TCID50 titer of RABV in the three kinds of glands of the mice and the copy number of the RABV N gene were de-tected,and RABV in different salivary glands was observed by immunofluorescence.The results showed that PB4 was dispersed in all three kinds of salivary glands of the mice,with the largest a-mounts in the parotid gland,followed by the submandibular gland,and the lowest amount in the sublingual gland.Three-month-old dogs were inoculated with PB4 through the cranial cavity,and saliva were collected every 12 h after inoculation.The saliva samples were detected by TCID50 and RT-qPCR.And during the moribund stage of the dogs when the disease occurred,the three pairs of salivary glands were collected.Through the determination of the TCID50 titer,RT-qPCR and immu-nofluorescence detection,it was demonstrated that among the three different salivary glands of the dogs,the largest amount of PB4 was found in the parotid gland and the lowest in the sublingual gland.Our results in mice and dogs clearly proved that the parotid gland was consistently found to exhibit the highest content of street RABV among the three major salivary glands,which could en-rich experimental data for analyzing the dispersion of RABV in the salivary glands and interpreta-tion of the intermittent secretion of saliva in clinically rabid dogs.

This study was aimed at exploring the prevalence and drug sensitivity of Gordonia strains isolated from sputum samples in Henan Province,to provide data to aid in the prevention and treatment of Gordonia infection.A combination of 16S rDNA and sec A1 gene sequencing was used to identify the isolated strains,and susceptibility to16 drugs was determined with the broth microdilution method.A total of 21 strains were identified through 16S rDNA gene and sec A1 gene sequencing,including five strains of Gordonia broncians,eight strains of Gordonia paraphernivans,seven strains of Gordonia sputi,and one strain of Gordonia aichiensis.Drug sensi-tivity testing showed high Gordonia sensitivity to drugs such as ceftriaxone,linezolid,doxycycline,amoxicillin/clavulanic acid,mino-cycline,cefotaxime,trimethoprim/sulfamethoxazole,imipenem,tobramycin,and clarithromycin.The sensitivity rates of the isolated strains were 90.48%(19/21),100%(21/21),90.48%(19/21),90.48%(19/21),95.24%(20/21),90.48%(19/21),90.48%(19/21),90.48%(19/21),and 95.24%(20/21),respectively.Gordonia showed high resistance to rifampicin and cefepime,with rates of 28.57%(6/21)and 19.05%(4/21),respectively.Meanwhile,the resistance varied among bacterial strains.The resistance rate of G.sputi to rifampicin reached 71.43%(5/7),whereas that of G.parapffinivoras to cefepime was 37.5%(3/8).The main species of Gordo-nia isolated from sputum samples of patients in Henan Province were G.bronchialis,G.paraffinivoras,G.sputi,and G.aichiensis.Drug sensitivity tests indicated that drugs including amoxicillin/clavulanic acid,ceftriaxone,cefotaxime,tobramycin,clarithromycin,mi-nocycline,trimethoprim/sulfamethoxazole,linezolid,and doxycycline had good antibacterial effects against Gordonia.

Rabies is a zoonotic disease that poses a global public health threat.Rabies virus(RABV)is neurotropic and can cause severe neurological disorders and behavioral abnormalities in host,with a fatality rate nearly 100％.In order to identify the key genes for RABV affecting neuronal cell function,we established and analyzed the mRNA expression profile of Neuro-2a(N2a)cell in-fected with challenge virus standard(CVS)-11 by RNA sequencing(RNA-seq).Biological func-tions of differentially expressed genes(DEGs)were determined by GO and KEGG enrichment a-nalysis.The results showed that there were 415 differentially expressed genes in N2a infected with CVS-11 strain,of which 89 were up-regulated and 326 were down-regulated.These genes were re-lated to a variety of biological processes,such as axon guidance pathway,cholesterol metabolism pathway,nitrogen metabolism pathway,and glycosphingolipid biosynthesis pathway,many of them have been shown to be closely associated with RABV infection.A total of 12 DEGs related to axon conduction,antigen processing and presentation pathways were selected and detected by real-time PCR,and their expression trends were consistent with the RNA-seq results.The genomic tran-scriptomic data on N2a cell under RABV infection will provide new clues for probing the mecha-nisms of RABV infection and transmission in the nervous system in the future.

Objective:To explore the efficacy, adverse reactions, and feasibility of high-definition transcranial direct current stimulation(HD-tDCS) treating major depressive disorder(MDD) patients with anxious distress.Methods:Sixty cases of MDD with anxious distress admitted to Zhenjiang Mental Health Center were recruited as participants. All patients were allocated into either the active treatment group or the sham group based on the random number table method. HD-tDCS was utilized in both groups to stimulate the left dorsolateral prefrontal cortex on the basis of conventional antidepressant treatment. In the active group, 2mA current stimulation was used for 20 min, and in the sham group, a 30-sceond stimulation was adopted at the beginning and the end of the stimulation. Participants in both group were treated once a day, five times a week for two consecutive weeks. Anxiety and depression symptoms of the patients were assessed respectively by the Hamilton depression scale(HAMD 17), the Montgomery-Asberg Depression Rating Scale(MARDS), the Hamilton Anxiety Scale(HAMA), and the Beck Anxiety Inventory(BAI) at the baseline, 2nd, 4th, and the 6th weekend of the treatment. The differences between active treatment and sham groups were analyzed by repetitive measure analysis of variance and simple effect analysis. Fisher′s exact probability test was used to compare the effective rate, remission rate and adverse reaction rate between the two groups. Results:(1) The interaction of times and groups was significant in HAMD 17( F=3.29, P<0.05) and BAI( F=2.99, P<0.05). In all measurement instruments, the main effects of groups( F=4.40-7.94) and times( F=35.42-247.59) were significant( P<0.05).(2) Findings from the simple effect analysis showed that: there were no significant differences in the scores of HAMD 17 at baseline and the 2 nd assessment between the two groups. Similarly, no significant differences were found in BAI at baseline, the 2nd and the 6th weekend of the treatment between two groups( P>0.05).(3) The scores of HAMD 17 at the 4th and the 6th week, BAI at the 4th weekend of the treatment in the active group were significantly lower than that in the sham group( P<0.05).(4) At the 4th weekend of the treatment, the active group had a remission rate of 16/20 and a response rates of 19/20, which were higher than 9/19 and 13/19, respectively, in the sham group( P<0.05). Also, the remission rate(18/20) in the active group was higher than that in the sham group(11/19) at the 6th weekend of the treatment.( P<0.05). As for the response rates, differences were not found between the two groups at the 6th weekend of the treatment.(5) The overall dropout rate had no significant differences in-between( P>0.05). As for the safety outcome, the rate of adverse events(e.g., itching, tingling and headache) showed no significant differences between the two groups. Additionally, no severe adverse events or mania was reported. Conclusions:This study indicated that HD-tDCS has significant efficacy and high safety in the treatment among MDD patient with anxious distress. Nevertheless, further large sample clinical studies are warranted to confirm the findings of the current investigation.

Objective:To explore the efficacy, adverse reactions, and feasibility of high-definition transcranial direct current stimulation(HD-tDCS) treating major depressive disorder(MDD) patients with anxious distress.Methods:Sixty cases of MDD with anxious distress admitted to Zhenjiang Mental Health Center were recruited as participants. All patients were allocated into either the active treatment group or the sham group based on the random number table method. HD-tDCS was utilized in both groups to stimulate the left dorsolateral prefrontal cortex on the basis of conventional antidepressant treatment. In the active group, 2mA current stimulation was used for 20 min, and in the sham group, a 30-sceond stimulation was adopted at the beginning and the end of the stimulation. Participants in both group were treated once a day, five times a week for two consecutive weeks. Anxiety and depression symptoms of the patients were assessed respectively by the Hamilton depression scale(HAMD 17), the Montgomery-Asberg Depression Rating Scale(MARDS), the Hamilton Anxiety Scale(HAMA), and the Beck Anxiety Inventory(BAI) at the baseline, 2nd, 4th, and the 6th weekend of the treatment. The differences between active treatment and sham groups were analyzed by repetitive measure analysis of variance and simple effect analysis. Fisher′s exact probability test was used to compare the effective rate, remission rate and adverse reaction rate between the two groups. Results:(1) The interaction of times and groups was significant in HAMD 17( F=3.29, P<0.05) and BAI( F=2.99, P<0.05). In all measurement instruments, the main effects of groups( F=4.40-7.94) and times( F=35.42-247.59) were significant( P<0.05).(2) Findings from the simple effect analysis showed that: there were no significant differences in the scores of HAMD 17 at baseline and the 2 nd assessment between the two groups. Similarly, no significant differences were found in BAI at baseline, the 2nd and the 6th weekend of the treatment between two groups( P>0.05).(3) The scores of HAMD 17 at the 4th and the 6th week, BAI at the 4th weekend of the treatment in the active group were significantly lower than that in the sham group( P<0.05).(4) At the 4th weekend of the treatment, the active group had a remission rate of 16/20 and a response rates of 19/20, which were higher than 9/19 and 13/19, respectively, in the sham group( P<0.05). Also, the remission rate(18/20) in the active group was higher than that in the sham group(11/19) at the 6th weekend of the treatment.( P<0.05). As for the response rates, differences were not found between the two groups at the 6th weekend of the treatment.(5) The overall dropout rate had no significant differences in-between( P>0.05). As for the safety outcome, the rate of adverse events(e.g., itching, tingling and headache) showed no significant differences between the two groups. Additionally, no severe adverse events or mania was reported. Conclusions:This study indicated that HD-tDCS has significant efficacy and high safety in the treatment among MDD patient with anxious distress. Nevertheless, further large sample clinical studies are warranted to confirm the findings of the current investigation.

To investigate the effect of miR-802 on insulin secretion by islet β cells and its mechanism, miR-802 was overexpressed or knocked down in primary islet cells and Min6 cells via transfecting miR-802 mimic and miR-802 inhibitor, respectively. The effect of miR-802 on insulin secretion was detected by ELISA. The target gene of miR-802 was confirmed by miRNA target gene database prediction, luciferase report and Western blot. The function recovery experiment was carried out to clarify the mechanism of miR-802 regulating β cell secretion of insulin. The results showed that overexpression of miR-802 in islet primary cells and Min6 cells inhibited insulin secretion. qPCR and Western blot showed that miR-802 inhibited insulin secretion by inhibiting the transcription and translation of the target gene, hepatocyte nuclear factor 1β(Hnf1B).

Objective To investigate the effects of dorsal hippocampal lesions (DH) or fimbria-fornix transection (FF) on the learning and memory of conditioned fear and the heart rate and blood pressure in rats.Methods Nineteen male adult Wistar rats were used in this experiment.They were implanted telemetry sensors in their abdominal aortas.Two week later,six of the rats were subjected to permanent NMDA-induced neurotoxic lesions to the dorsal hippocampus (DH) and seven for the fimbria-fornix transection (FF)through stereotactic brain surgery,the left six were treated with saline as the control (Sham).All rats were subjected to a conditioned fear experiment.Meanwhile,changes in heart rate and blood pressure were measured.Results There was no significant difference in heart rate and blood pressure among the rats with the hippocampal operation or fimbria-fornix transection.In the acquisition of conditioned fear,there were significant difference in freezing time among the three group in both inter-trial-interval (ITI) and conditioned stimulus (CS) process (all P＜0.05).The freezing time of the FF group showed significantly lower than that of the Sham group (P＜0.05).There was no significant difference in heart rate and blood pressure among the three group(P＞0.05).In the test of conditioned contextual fear memory,the freezing time percentage in the FF group ((0.31±0.16) ％) significantly lower than that in the Sham group ((2.78± 1.23) ％) (P＜0.05)at the first 3 min of the test.There was a significant difference in heart rate among the three group.The heart rate of FF group ((436.42± 10.16) times/min) was significantly lower than that of the Sham group ((472.48±4.43) times/min,P＜0.01) and the DH group ((469.94 ±7.36)times/min,P＜0.01).In the test of conditioned tone fear memory.The freezing time percentage in FF group ((18.78±6.29) ％) was significantly lower than that in the Sham ((51.77±9.33)％,P＜0.01) and DH group ((59.19±8.13)％,P＜0.01),but the freezing time percentage between the later two groups had no difference (P=0.52).The synchronous telemetry measurement showed there was no significant difference both in the heart rate and the blood pressure among the groups (all P＞0.05) during the conditioned tone test.Conclusion The dorsal hippocampal lesions and fimbria-fornix transection in rats can significantly reduce the learning and memory ability in conditioned fear and scene fear in rats,and the effect of fimbria-fornix transection is more obvious.The decrease in,fear memory is not synchronously reflected in heart rate and blood pressure in rats.

Objective To investigate the genotype of M.tuberculosis in He'nan Province.Methods A total of 668 M.tuberculosis clinical strains collected in difference regions of He'nan Province during 2015 were genotyped by two standard methods,including classical 24-locus mycobacterium interspersed repetitive unit variable-number tandem-repeat (MIRU-VNTR) typing and spoligotyping.Results The 668 isolates were divided into 11 clusters and 35 patterns by spoligotyping.Among the 558 Beijing strains,546 were typical Beijing strains and the other 12 were atypical Beijing strains.Among the 110 non-Beijing strains,eight were new strains and the remaining 102 non-Beijing strains were divided into 10 families.There were 76 isolates belonging to T family,including 59 of T1 families,7 of T2 families,and 10 of T3 families.The 668 strains were divided into 550 gene patterns by standard 24-locus VNTR,including 508 un-clustered patterns and 160 clustered into 42 clusters.The largest cluster contained 21 strains,the other clusters contained 2-20 strains.Conclusion Beijing strain is still the most prevalent M.tuberculosis in He'nan Province.