Long-Term Exposure to Ceftriaxone Sodium Induces Alteration of Gut Microbiota Accompanied by Abnormal Behaviors in Mice.

Background: Growing evidence points out that a disturbance of gut microbiota may also disturb the gut-brain communication. However, it is not clear to what extent the alteration of microbiota composition can modulate brain function, affecting host behaviors. Here, we investigated the effects of gut microbiota depletion on emotional behaviors. Methods: Mice in the experimental group were orally administered ceftriaxone sodium solution (250 mg/ml, 0.2 ml/d) for 11 weeks. The open-field test and tail-suspension test were employed for the neurobehavioral assessment of the mice. Fecal samples were collected for 16s rDNA sequencing. The serum levels of cytokines and corticosterone were quantified using enzyme-linked immunosorbent assays. The immunohistochemistry method was used for the detection of brain-derived neurotrophic factor (BDNF) and c-Fos protein. Results: The gut microbiota for antibiotic-treated mice showed lower richness and diversity compared with normal controls. This effect was accompanied by increased anxiety-like, depression-like, and aggressive behaviors. We found these changes to be possibly associated with a dysregulation of the immune system, abnormal activity of the hypothalamic-pituitary-adrenal axis, and an alteration of neurochemistry. Conclusions: The findings demonstrate the indispensable role of microbiota in the gut-brain communication and suggest that the absence of conventional gut microbiota could affect the nervous system, influencing brain function.

Long non­coding RNA CASC2 enhances berberine­induced cytotoxicity in colorectal cancer cells by silencing BCL2.

Berberine, a natural isoquinoline alkaloid derived from Berberis species, has been reported to have anticancer effects. However, the mechanisms of action in human colorectal cancer (CRC) are not well established to date. In the present study, the cell cytotoxicity effect of berberine on human CRC cells, as well as the possible mechanisms involved, was investigated. The results of the cell viability and apoptosis assay revealed that treatment of CRC cells with berberine resulted in inhibition of cell viability and activation of cell apoptosis in a concentration­dependent manner. To reveal the underlying mechanism of berberine­induced anti­tumor activity and cell apoptosis, RNA­sequencing followed by reverse­transcription quantitative PCR were performed. In addition, RNA immunoprecipitation, chromatin immunoprecipitation and western blot analysis were used to identify the functional regulation of CASC2/EZH2/BCL2 axis in berberine­induced CRC cell apoptosis. The data revealed that lncRNA CASC2 was upregulated by berberine treatment. Gain­ or loss­of­function assays suggested that lncRNA CASC2 was required for the berberine­induced inhibition of cell viability and activation of cell apoptosis. Subsequently, the downstream antiapoptotic gene BCL2 was identified as a functional target of the berberine/CASC2 mechanism, as BCL2 reversed the berberine/CASC2­induced cell cytotoxicity. lncRNA CASC2 silenced BCL2 expression by binding to the promoter region of BCL2 in an EZH2­dependent manner. In summary, berberine may be a novel therapeutic agent for CRC and lncRNA CASC2 may serve as an important therapeutic target to improve the anticancer effect of berberine.

Berberine Promotes Apoptosis of Colorectal Cancer via Regulation of the Long Non-Coding RNA (lncRNA) Cancer Susceptibility Candidate 2 (CASC2)/AU-Binding Factor 1 (AUF1)/B-Cell CLL/Lymphoma 2 (Bcl-2) Axis.

BACKGROUND Berberine, a natural isoquinoline alkaloid derived from Berberis genus plants, has been reported to have anti-cancer effects. While cell behavior can be modulated by long non-coding RNAs (lncRNAs), the contributions of lncRNAs in progression and berberine effects on colorectal cancer are largely unknown. Therefore, the present study investigated the involvement and regulatory function of lncRNA cancer susceptibility candidate 2 (CASC2) during the treatment of human colorectal cancer using berberine. MATERIAL AND METHODS Reverse transcription-quantitative PCR (RT-qPCR) was performed to detect the expression levels of lncRNA CASC2 and Bcl-2 mRNA in colorectal cancer cells. MTT assay was performed to evaluate cell viability. Flow cytometry and TUNEL assay were used to analyze the apoptosis of cancer cells. RNA immunoprecipitation (RIP) assay was done to verify the interaction between lncRNA CASC2 and (AU-binding factor 1) AUF1, or AUF1 and B-cell CLL/lymphoma 2 (Bcl-2). RESULTS Treatment with berberine suppressed cell viability of colorectal cancer by promoting apoptosis level. LncRNA CASC2 was upregulated in cells treated with berberine, and knockdown of lncRNA CASC2 reversed the berberine-induced apoptosis. In addition, anti-apoptotic gene Bcl-2 was suppressed by berberine treatment and lncRNA CASC2, inducing the pro-apoptotic effects. Moreover, lncRNA CASC2 binds to AUF1, which sequestered AUF1 from binding to Bcl-2 mRNA, thus inducing the inactivation of Bcl-2 translation. CONCLUSIONS Our study reveals that lncRNA CASC2 mediates the berberine-induced pro-apoptotic effect via inhibition of Bcl-2 expression at the post-transcriptional level.

[Epidemiology and Resistance Mechanisms of Group B Streptococci in Late-pregnant Maternal Birth Canal].

OBJECTIVE: To study the antibiotic-resistant rate of group B streptococci (GBS) in obstetric canal of late-pregnant women, evaluate the antibiotic-resistant status and finally to support the GBS prevention and curing by proper antibiotics. METHODS: 31 pregnant women between 35 to 37 gestational weeks were included, for whom the antibiotic sensitivity as well as the drug (erythromycin and clindamycin) resistance genes of GBS in obstetric canal was analyzed. RESULTS: 12 (38. 7%) strains of GBS were resistant to clindamycin, while 21 (67. 7%) to erythromycin, within which 12 strains were intrinsic phenotype - cMLS type-clindamycin resistance, other 9 were active efflux phenotype - MS type-clindamycin sensitive and all of which were confirmed by Double disk diffusion method. Eleven strains were mef (A) positive, and 12 strains were erm (B) positive, in which 3 with erm (C). CONCLUSIONS: In our research the GBS strains show a high erythromycin and clindamycin resistance rate. The resistance of our GBS strains are mainly caused by the ribosomal target changes induced by erm (B) and the increased efflux of clindamycin induced by mef (A).

[Clinical characteristics of children with meningitis caused by Streptococcus pneumoniae and antibiotic resistance of Streptococcus pneumoniae strains].

OBJECTIVE: To study the clinical characteristics of children with meningitis caused by Streptococcus pneumoniae (SP) and the drug sensitivity of SP strains. METHODS: A retrospective analysis was performed on the clinical data of 14 children with SP-infected meningitis between September 2008 and March 2014. RESULTS: Of the 14 cases, 8 cases (57%) aged under 2 years. 13 cases (93%) had fever, 9 cases (64%) had convulsions, and 7 cases (50%) were complicated by septicemia. Eleven cases (79%) had elevated white blood cell (WBC) counts and 10 cases (71%) had elevated serum C-reactive protein (CRP) levels. All 14 children had an elevated nucleated cell count and neutrophils were identified as the predominant cell type. CSF protein>1000 mg/dL was noted in 9 cases (64%). Ten cases (71%) were cured, 2 cases (14.2%) with sequelae and 2 cases (14.2%) died. The drug sensitivity analysis showed that SP had resistance rates of more than 60% to penicillin, erythromycin, clindamycin, tetracycline and sulfa, but it was sensitive to amoxicillin (93%), vancomycin (100%), chloramphenicol (100%) and levofloxacin (100%). CONCLUSIONS: The clinical characteristics of children with meningitis caused by SP are not different from those with meningitis caused by other bacteria. SP strains are resistant to common antibiotics used in clinical practice, so it is important to monitor the drug resistance of the strains.

Identification of cytolytic vaginosis versus vulvovaginal candidiasis.

OBJECTIVE: This study aimed to observe the morphological characteristic of vaginal discharge in patients with cytolytic vaginosis (CV) under the microscope and to identify it in patients with CV and in patients with vulvovaginal candidiasis (VVC). METHODS: A total of 108 subjects including 21 healthy women, 33 patients with CV, and 54 patients with VVC were enrolled in the present morphological study. Vaginal discharge was collected and made into smear. The morphological characteristics of these vaginal smears with Gram staining were observed under the microscope. The smears were assessed for the quantity of lactobacilli, epithelial cell morphology, and the absence or presence of Candida species, Trichomonas vaginalis, and clue sells. RESULTS: First, the age, the level of education, and especially the status of pregnancy of patients with CV were significantly different from those of the patients with VVC. Second, the morphological characteristics of patients with CV consisted of overgrowth of lactobacilli, the presence of naked nuclei and fragments of the epithelial cells, a paucity of leukocytes, and the absence of Candida species and other pathogens. However, the morphological characteristic of patients with VVC consisted of the presence or absence of lactobacilli and the presence of normal epithelial cells, candidal spores, blastospores, hyphae, or other pathogens such as T. vaginalis and Gardnerella vaginalis. CONCLUSIONS: Both CV and VVC can be identified based on the quantity of lactobacilli, the morphology of the epithelial cells, and the absence or presence of Candida species and other pathogens, and the misdiagnosis of CV as VVC can be avoided.

[Association of serum high-density lipoprotein cholesterol level and risk of recurrent ischemic stroke].

OBJECTIVE: To observe the association between high-density lipoprotein cholesterol (HDL-C) level and rate of ischemic stroke recurrence. METHODS: A total of 1 059 patients with ischemic stroke were enrolled from 5 community health centers and underwent baseline surveys during the period of January 2003 to December 2006. After baseline surveys, patients were followed up every 6 months until December 31, 2008. The new stroke events were recorded as the primary study endpoint. The association between HDL-C, HDL-C/TC and ischemic stroke recurrence was analyzed by Cox regression analysis. RESULTS: The proportions of stroke patients with high ( ≥ 1.55 mmol/L), moderate (1.04-1.54 mmol/L) and low (<1.04 mmol/L) HDL-C levels were 15.58% (165/1 059) , 54.58% (578/1 059) and 29.84% (316/1 059) respectively. During a mean of (3.21 ± 1.04) years follow-up, recurrent ischemic stroke was recorded in 137 patients. Compared with HDL-C ≥ 1.40 mmol/L group, multivariate Cox regression analysis showed that stroke recurrence rates of patients with HDL-C ≤ 1.00 mmol/L and ranged from 1.01 to 1.19 mmol/L increased by 0.944 (HR = 1.944, 95%CI:1.033-3.659, P = 0.039) and 1.027 (HR = 2.027, 95%CI:1.116-3.682, P = 0.020)fold , respectively. Recurrence rates increased 1.237 (HR = 2.237, 95%CI:1.208-4.144, P = 0.010) fold in patients with HDL-C/TC ≤ 0.19 mmol/L compared to patients with HDL-C/TC ≥ 0.28 mmol/L. CONCLUSION: The risk of ischemic stroke recurrence increases with decreasing HDL-C level or HDL-C/TC ratio.

[Species and drug resistance of pathogens in blood cultures from the pediatric hematology ward].

OBJECTIVE: To investigate the species and percentage changes of pathogens in blood cultures from the pediatric hematology ward, and to analyze the drug resistance of main pathogens and the risk factors for positive blood culture (sepsis). METHODS: A retrospective analysis was performed to analyze the species and drug sensitivity of the pathogens isolated from 2358 blood cultures from the pediatric hematology ward of the West China Second University Hospital between 2008 and 2011, as well as the related clinical data. RESULTS: A total of 110 strains of pathogens were isolated, with Escherichia coli (16 strains), Pseudomonas aeruginosa (12 strains) and Staphylococcus epidermidis (8 strains) being the most common ones. From 2008 to 2011, the percentage of Gram-positive bacteria decreased, while the percentage of Gram-negative bacteria increased. The detection rates of methicillin-resistant coagulase-negative Staphylococci and methicillin-resistant Staphylococcus aureus were 69% and 43% respectively, but both were sensitive to vancomycin. The detection rates of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and ESBL-producing Klebsiella pneumoniae were 69% and 62% respectively, but both were sensitive to imipenem and meropenem. Malignant tumor was a risk factor for positive blood culture (OR=3.564, P<0.05). CONCLUSIONS: A wide range of pathogens are responsible for bloodstream infection in the pediatric hematology ward and the percentages of bacteria are changing; these pathogens have a high drug resistance rate. Malignant tumor is a risk factor for positive blood culture in the pediatric hematology ward.

[Common pathogens and clinical characteristics of neonatal pneumonia].

OBJECTIVE: To study common pathogens and their antibiotic susceptibility as well as clinical characteristics of neonatal pneumonia. METHODS: A retrospective study on neonatal pneumonia was performed. The study investigated antibiotic susceptibility of four common pathogens (339 strains) that caused neonatal pneumonia. Clinical characteristics of the newborns with pneumonia were analyzed. Of the 339 strains, 185 were isolated from bronchial secretions, 72 from blood samples, and 82 with positive results of both samples. RESULTS: Four hundred and seventy-four neonates with pneumonia presented positive results of bacterial culture. the most common pathogens Staphylococcus aureus (21.9%), Escherichia coli (19.2%), Klebsiella pneumoniae (19.0%) and Enterobacter cloacae (11.4%). The birth weight of newborns infected with Staphylococcus aureus was generally normal, and the time of hospital admission was later (after 24 hours of life). In contrast, the newborns with gram-negative bacterial infection, especially Klebsiella pneumoniae infection, had lower birth weights and early time of hospital admission (within 24 hours of life). Nearly more than 50% gram-negative bacteria were resistant to second, third and forth generation cephaloporins. CONCLUSIONS: Gram-negative bacteria are predominant pathogens of neonatal pneumonia. Neonatal pneumonia caused by gram-negative bacteria is common in newborns with low birth weight and its onset time is relatively earlier. Gram-negative bacteria that cause neonatal pneumonia are highly resistant to cephaloporins.

[Clinical characteristics and pathogens of invasive fungal infections in children].

OBJECTIVE: To study the clinical characteristics and pathogens of invasive fungal infection in children. METHODS: The clinical data of 104 children who suffered from invasive fungal infections between 2008 and 2012 was retrospectively reviewed. RESULTS: Of the 104 cases, 20 occurred in neonates, 48 in infants and 36 in preschool and school-aged children (old-aged children). Prematurity (70%), hyaline membrane disease (45%) and pneumonia (30%) were commonly comorbid in the neonate group. In addition, the percentage of cases receiving total parenteral nutrition was higher in the neonate group than in the other two age groups (P<0.01). Mechanical ventilation was more frequent in neonate and infant groups than in the old-aged children (P<0.01). Hematological malignancy was the most common underlying disease, and the percentage of children who had neutropenia and accepted chemotherapy was higher in the old-aged children than in the other two age groups (P<0.05). Lung infection was the most common (61.5%), followed by sepsis (14.4%) and intestinal tract infection (12.5%), while nervous system infections were found only in old-aged children. A total of 105 strains of fungi were isolated from the 104 patients, including Candida (n=90, 85.7%), Cryptococcus (n=6) and others (n=9). The most commonly isolated species was Candida albicans (n=52, 49.5%). Non-Candida albicans Candida accounted for 36.2% (n=38). The rate of susceptibility of Candida species to 5-fluorocytosine and amphotericin B was higher than fluconazole. CONCLUSIONS: Invasive fungal infections can occur in children at various ages. There are differences in the risk factors for invasive fungal infections between age groups. Candida species are the main pathogens of childhood invasive fungal infections, and both Candida albicans and non-Candida albicans Candida are common. Fluorocytosine and amphotericin B are sensitive antifungal agents for infections caused by Candida species.

[Metabolic syndrome is an independent risk factor for cardiovascular disease events in patients with ischemic stroke].

OBJECTIVE: To explore the association between metabolic syndrome (MS) and risk of cardiovascular disease events (CVD) in patients with ischemic stroke. METHOD: A total of 1087 patients with ischemic stroke were enrolled from 5 community-based medical centres and underwent baseline evaluation on risk factors of stroke during the period of Jan. 2003 to Dec. 2006. After baseline survey, all patients were followed up until Dec 31, 2008 and new CVD events were recorded. MS was defined using CDS criteria. Proportional hazard models were used to assess the HRs and 95% CI of CVD events associated with MS and other components. RESULTS: The prevalence of MS was 40.4% at baseline. During an average follow-up of 3.5 years, 178 patients developed new CVD events. After adjusted for age, gender, smoking, drinking, marriage status, education level, hospitalization, recurrence of stroke, stroke duration, depression, cognition impairment and ADL, MS remains the independent predictor for the risk of CVD events. Compared with patients with non-MS, the risk of CVD events increased by 44% (HR: 1.44, 95%CI: 1.06 - 1.95). The risk of CVD also increased with the number of MS components. Compared with patients with 1 or less than 1 components of MS, the risk of CVD events increased by 30% (HR: 1.30, 95% CI: 0.83 - 2.04) in those with 2 components and by 69% (HR: 1.69, 95%CI: 1.11 - 2.56) in those with 3 or more components of MS. Hypertension and hyperglycemia and impaired fasting glucose also served as independent risk factors for CVD event (all P < 0.001). CONCLUSIONS: MS was independently associated with increased risk of CVD events in patients with ischemic stroke. There was a dose-response relationship between the numbers of MS components and the risk of CVD event.