Paenibacillus nasutitermitis sp. nov., isolated from a termite gut.

A Gram-stain-positive, non-motile, aerobic and terminal-endospore-forming rod-shaped bacterium, strain P5-1T, was isolated from the hindgut of a wood-feeding higher termite, Nasutitermes sp. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain was closely related to Paenibacillus sepulcri CCM 7311T (97.5 % similarity). Growth was observed at 10-40 °C (optimum, 30 °C) and at pH 5.5-9.0 (optimum, pH 7.5). The DNA G+C content of strain P5-1T was 48.9 mol%. Cells contained menaquinone 7 (MK-7) as the sole respiratory quinone and the major fatty acids were anteiso-C15 : 0 and iso-C15 : 0. The cellular polar lipids comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, one unidentified phospholipid, one unidentified glycolipid and one unidentified aminophospholipid. The diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid. Based on the phylogenetic, chemotaxonomic and phenotypic data obtained within this study, strain P5-1T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus nasutitermitis sp. nov. is proposed. The type strain is P5-1T ( = CGMCC 1.15178T = NBRC 111536T).

[Prevention of Inonotus obliquus polysaccharides for high power microwave radiation induced testicular injury in rats: an experimental research].

OBJECTIVE: To investigate the effect of Inonotus obliquus polysaccharides on testicular injury induced by exposure to high power microwave (HPM) in rats. METHODS: A total of 30 male Wistar rats were randomly divided into 5 groups, i.e., the normal control group, the microwave radiation model group, the treatment group, the new microwave radiation model group, and the prevention group, 6 in each group. All rats, except those in the normal control group, were exposed to microwave at an average power density of 200 mW/cm2 for 6 min. Rats in the control group and the model group were administered with normal saline by gastrogavage, once a day. Rats in the treatment group and the prevention group were given with Inonotus obliquus polysaccharides by gastrogavage, 2 mL each time (400 mg/kg body weight), once a day. All rats were sacrificed on the 11th day.The sperm density and the rate of sperm deformity were determined. Pathological changes of testis were observed by light microscope and transmission electron microscope. RESULTS: Short-term HPM irradiation could significantly reduce the sperm density and increase the sperm deformity rate (P < 0.05). Meanwhile, obvious pathological changes of testes occurred. Compared with the two model groups, the sperm density increased and the sperm deformity rate decreased in the treatment group and the prevention group (P < 0.05). Under the light microscope, injuries of spermatogenic cells and stromal cells, as well as vascular dilatation and congestion were obviously alleviated in the treatment group and the prevention group. Mitochondrial swelling and endoplasmic reticulum expansion shown by ultrastructural observation were also significantly alleviated. Of them, injuries of spermatogenic cells and inflammation response were milder in the treatment group than in the prevention group. CONCLUSIONS: Inonotus obliquus polysaccharides had significant protective effect on microwave radiation induced testicular injury. Better effect was obtained by therapeutic medication than preventive medication.

Hsa_circ_0001402 alleviates vascular neointimal hyperplasia through a miR-183-5p-dependent regulation of vascular smooth muscle cell proliferation, migration, and autophagy.

INTRODUCTION: Vascular neointimal hyperplasia, a pathological process observed in cardiovascular diseases such as atherosclerosis and pulmonary hypertension, involves the abundant presence of vascular smooth muscle cells (VSMCs). The proliferation, migration, and autophagy of VSMCs are associated with the development of neointimal lesions. Circular RNAs (circRNAs) play critical roles in regulating VSMC proliferation and migration, thereby participating in neointimal hyperplasia. However, the regulatory roles of circRNAs in VSMC autophagy remain unclear. OBJECTIVES: We aimed to identify circRNAs that are involved in VSMC autophagy-mediated neointimal hyperplasia, as well as elucidate the underlying mechanisms. METHODS: Dual-luciferase reporter gene assay was performed to validate two competing endogenous RNA axes, hsa_circ_0001402/miR-183-5p/FKBP prolyl isomerase like (FKBPL) and hsa_circ_0001402/miR-183-5p/beclin 1 (BECN1). Cell proliferation and migration analyses were employed to investigate the effects of hsa_circ_0001402, miR-183-5p, or FKBPL on VSMC proliferation and migration. Cell autophagy analysis was conducted to reveal the role of hsa_circ_0001402 or miR-183-5p on VSMC autophagy. The role of hsa_circ_0001402 or miR-183-5p on neointimal hyperplasia was evaluated using a mouse model of common carotid artery ligation. RESULTS: Hsa_circ_0001402 acted as a sponge for miR-183-5p, leading to the suppression of miR-183-5p expression. Through direct interaction with the coding sequence (CDS) of FKBPL, miR-183-5p promoted VSMC proliferation and migration by decreasing FKBPL levels. Besides, miR-183-5p reduced BECN1 levels by targeting the 3'-untranslated region (UTR) of BECN1, thus inhibiting VSMC autophagy. By acting as a miR-183-5p sponge, overexpression of hsa_circ_0001402 increased FKBPL levels to inhibit VSMC proliferation and migration, while simultaneously elevating BECN1 levels to activate VSMC autophagy, thereby alleviating neointimal hyperplasia. CONCLUSION: Hsa_circ_0001402, acting as a miR-183-5p sponge, increases FKBPL levels to inhibit VSMC proliferation and migration, while enhancing BECN1 levels to activate VSMC autophagy, thus alleviating neointimal hyperplasia. The hsa_circ_0001402/miR-183-5p/FKBPL axis and hsa_circ_0001402/miR-183-5p/BECN1 axis may offer potential therapeutic targets for neointimal hyperplasia.

Correlation between an ABO Blood Group and Primary Femoral Head Necrosis: A Case-Control Study.

OBJECTIVE: To investigate the relationship between primary femoral head necrosis (ONFH) and an ABO blood group. METHODS: This study was a retrospective case-control trial. An analysis of the clinical data of an ABO blood group with 516 patients (case group) with ONFH and 489 limb-fracture patients (control group) without previous hip pain was obtained from the Second Hospital of Shanxi Medical University from November 2015 to November 2018. The clinical data included gender, age, height, weight, a history of smoking, alcohol abuse, prior medical history, hormone use, and ABO blood type. A logistic regression model was used for univariate and multivariate analysis. RESULTS: From November 2015 to November 2018, there were 267 males and 249 females in the 516 cases of ONFH in the case group. The control group included 289 males and 200 females. In terms of age, the average age of the case group was significantly lower than that of the control group. In terms of body mass index (BMI), the BMI of the case group was significantly higher than that of the control group (P < 0.05). From the previous medical history of patients in the two groups (coronary heart disease, hypertension, cerebrovascular disease, diabetes, and peripheral vascular disease), there was no significant difference between the two groups from a statistical perspective (P < 0.05). However, according to the risk factors of ONFH (smoking, alcohol abuse, hyperlipidemia, and hormone-use history), there were significant differences between the case group and the control group. There was no statistical difference in the quantitative distribution ratio of the four blood types - A, B, O, and AB - between the case group and the control group. The outcomes of logistic multiple regression analysis presented that there was no significant correlation between the occurrence of ONFH and blood type A, B, AB, and O (P > 0.05). However, there are significant differences in the disease progression between the different blood types. There was a significant difference in the progression of disease between type A and type O. Among them, patients with ONFH and type A blood had the fastest progression with an average of 2.318 years, and the slowest progression was found in type O blood with an average of 5.15 years. CONCLUSIONS: The ABO blood group has no correlation with the occurrence of ONFH, but the ABO blood type is closely related to the disease progression of ONFH.

Effect of Cholecystectomy on the Occurrence of Knee Osteoarthritis.

OBJECTIVE: To evaluate the effect of cholecystectomy on the occurrence of knee osteoarthritis (KOA). METHODS: The present study was a case-control study with a retrospective, cross-sectional, and longitudinal study design. The clinical data for knee osteoarthritis in the Second Hospital of Shanxi Medical University from January 2016 to September 2018 was analyzed. Clinical data, including gender, age, height, weight, smoking, alcohol abuse, prior medical history, and previous surgical history, were recorded. A logistic regression model was used for the univariate and multivariate analysis. RESULTS: A total of 1659 patients with KOA (KOA group) and 1195 limb fracture patients (control group) were included in this study. Among the 1659 patients with KOA, 388 patients were male and 1271 were female, while among the 1195 patients in the control group, 638 patients were male and 557 patients were female. The period between cholecystectomy and knee osteoarthritis onset ranged from 0.5 years to 17 years; the average interval time was 8.73 ± 2.11 years. The age at disease onset was significantly older when compared to the control group, while the body mass index in the KOA group was significantly higher when compared to the control group (P < 0.05). There were 97 patients undergoing cholecystectomy in the KOA group and there were 15 patients undergoing cholecystectomy in the control group. The proportion of cholecystectomy in the two groups was statistically significant. After the univariate analysis, there was a statistically significant difference in distribution between the two groups (P < 0.05). The multivariate logistic regression analysis revealed that there was a significant difference in the distribution of these two groups (P < 0.05), indicating that cholecystectomy is associated with the occurrence of KOA. CONCLUSION: There is a close relationship between cholecystectomy and KOA. However, the specific mechanism remains unknown and should be further researched.

Progress of research on Inonotus obliquus.

Inonotus obliquus has high nutritional and medicinal value, especially in treating malignant tumors, diabetes, cardiovascular disease and AIDS, attracting significant attention from scholars in recent years. In this paper, the biological characteristics, chemical composition and pharmacologic effects of Inonotus obliquus were summarized. And the applications in medicine and food were introduced. Future research on Inonotus obliquus was also discussed in order to make Inonotus obliquus obtain effective exploitation and satisfy people's demands.

[Adenovirus mediated angiostatin gene therapy for ovarian cancer: experiment with nude mice].

OBJECTIVE: To built an expression vector of angiostatin (AG) gene with recombinated replication defective adenovirus and investigate the therapeutic effect of human AG gene on ovarian cancer. METHODS: (1) Human AG K (1 - 3) cDNA was inserted into the vector pShuttle to build the recombinant plasmid pShttle-AG (K1-3). pAdeno-X-AG (K1-3) was built by double-cut and recombinated pShttle-AG (K1-3) to vector pAdeno-X, and then recombinant adenovirus was finally prepared by transfection of pAdeno-X-AG (K1-3) into to the human embryo kidney cells of the line 293. (2) Human ovarian cancer cells of the line SKOV3 were inoculated subcutaneously into nude mice of the line BALB/c nu/nu to establish model of human ovarian cancer. Then the mice were randomly divided into 3 groups to be injected with Ad = AG (K1-3), Ad-LacZ, or phosphate buffered saline (PBS) around the cancer every 5 days. The tumor size was measured every 5 days to calculate the tumor volume and tumor inhibition rate. Three days after the last injection the mice were killed. The tumor tissues, livers, and kidneys of the mice underwent immunohistochemistry to calculate the microvessel density (MVD) and expression of vessel endothelial growth factor (VEGF) and AG. RESULTS: The tumor volume and weight of the Ad-AG (K1-3) group were significantly less than those of the PBS and Ad-LacZ groups (all P < 0.01), however, there were not significant difference between the latter two groups (both P > 0.05). The expression levels of CD34 and VEGF of the Ad-AG (K1-3) group were both significantly lower than those of the PBS and Ad-LacZ groups (all P < 0.01), however (both P > 0.05). CONCLUSION: Human angiostatin mediated by adenovirus suppresses the angiogenesis and the growth of human ovarian cancer in the nude mice model, which suggests that it is promising in clinical application.

Marriage and outcomes of people with schizophrenia in rural China: 14-year follow-up study.

The influence of marriage on the long-term outcomes of schizophrenia is largely unknown. This study was to examine the impact of marriage on the 14-year outcomes and identify the correlates of marriage among persons with schizophrenia in rural community. All study participants with schizophrenia (n=510) were identified in 1994 in an epidemiological investigation of 123,572 people aged 15years and older and followed up in 2004 and 2008 in Xinjin County, Chengdu, China. The Patients Follow-up Schedule (PFS) was used in 2004 and 2008. The rate of follow-up in 2008 was 95.9%. Unmarried individuals in 1994 had higher rates of homelessness and suicide, and lower rate of survival in 2004 and 2008 than those married. In 14-year follow-up, unmarried individuals were more likely to be male, to have higher level of psychiatric symptoms and lower rate of full remission of illness, and to report lower level of work functioning, as well as with fewer family members and caregiver, and lower family economic status. The predictors of being married in 2008 included being married in 1994, shorter duration of illness, being female, and lower level of education. Being married is predictive of more favorable 14-year outcomes of persons with schizophrenia in the rural community. Given that marriage can be instrumental for enhancing family-based support and caregiving, as well as improving the community tenure of persons with schizophrenia, it is important to develop programs to enhance opportunity for persons with schizophrenia to get and stay married.

Extracellular matrix metalloproteinase inducer and matrix metalloproteinase-2 overexpression is associated with loss of hormone receptor expression and poor prognosis in endometrial cancer.

Extracellular matrix metalloproteinase inducer (CD147) and matrix metalloproteinase-2 (MMP-2) have been documented in various malignancies. CD147 is a member of the immunoglobulin superfamily, which promotes the production and release of MMPs in mesenchymal cells and tumor cells. MMP-2 has been extensively studied and is considered to be particularly important in cancer invasion and metastasis. However, studies investigating the expression and prognostic value of CD147 in endometrioid endometrial carcinoma (EEC) are limited. The present study analyzed the expression of CD147 and MMP-2 by immunohistochemistry in endometrial tissue samples from 107 patients with EEC and 30 patients with benign uterus myoma. The association between CD147 and MMP-2 expression and clinicopathological characteristics was evaluated. The results showed that the overexpression of MMP-2 was significantly associated with International Federation of Gynecology and Obstetrics stage (P=0.007), depth of invasion (P=0.037) and reduced expression of progesterone receptor (P=0.005). Kaplan-Meier analyses indicated that CD147 overexpression alone (P<0.05 for disease-specific survival) or in combination with MMP-2 (P<0.001 for disease-specific survival) was correlated with adverse prognosis in EEC patients. Multivariate analysis revealed that the combined overexpression of CD147 and MMP-2 was an independent prognostic factor for disease-specific survival (hazard ratio=5.141, P=0.001) in EEC patients. CD147 and MMP-2 overexpression was positively correlated with aggressive phenotypic features in EEC, however it was negatively correlated with hormone receptor expression. The combination of CD147 and MMP-2 overexpression in EEC further distinguished a subgroup of patients with poor prognosis. Thus, the results of present study indicate that the co-expression of CD147 and MMP-2 may be an independent prognostic factor in EEC patients.

[Cloning and characterization of the promoter region of Musca domestica yolk protein-1 gene].

A partial Musca domestica genomic library was constructed. It was consisted of 1.2 x 10(5) recombinants with insert length ranging from 10 kb to 23 kb(15 kb average). High molecular weight genomic DNA with more than 50 kb size was extracted from the larva hatched 36 h and digested with unfrequently cutting restriction enzyme Bcl I. DNA fragments of 10 approximately 23 kb were recovered by agarose gel electrophoresis and ligated with EMBL3 BamH I Arms CIPase treated. Then the products of ligation were packed in vitro using packing protein. The cloning efficiency of the genomic library was 5 x 10(4) pfu/mL. The genomic library was screened by hybridization using a probe of a 768 bp partial cDNA fragment of Musca domestica yolk protein 1 (mdYP1) gene obtained by PCR and the probe was labeled with Digoxigen. A positive plaque was chosed and purified by in situ hybridization. A genomic DNA fragment about 4.0 kb mdYP1 was isolated from purified positive plaque by southern blotting analysis. Sequence analysis revealed that mdYP1 genomic gene was composed of 5'-upstream region about 1.7 kb with typical CAAT/TATA box. The promoter of the mdYP1 gene was characterized by examining the ability of 5'-upstream fragments to regulate expression of green fluorescent protein (GFP) in Musca domestica larva. Four fragments of the promoter region, P1 (+296/+7), P2 (+684/+7), P3(+1165/+7) and P4 (+1616/+7) ,were obtained by PCR specific amplification using template of recombinant A-lambdaNA containing mdYP1 gene sequence. Then the four fragments were respectively subcloned into pCMV-GFP reporter vector deleted CMV promoter. All the fragments showed no promoter activity when the four recombinant vectors were transfected into Sf9 and BHK -2 cells respectively, but three of them, P2, P3 and P4, showed significant promoter activity when they were respectively introduced into Musca domestica larva by electroporation. The two fragments, P5 (+684/+302) and P6 (+165/+302), obtained by digesting P2 and P3 with Spe I and Hind III, were also subcloned into pGFP vector, and they showed no promoter activity in Sf9 cells, BHK -21 cells and Musca domestica larva. The results demonstrated that the core promoter spanned 302bp and contained a CCAAT box and a TATA box upstream translation initiation codon (ATG), but itself had no transcriptional activity, and that regulatory promoters or enhancers and other cis-elements presented from +302 to +1616 were necessary to maintain the specific expression.

Inotodiol inhabits proliferation and induces apoptosis through modulating expression of cyclinE, p27, bcl-2, and bax in human cervical cancer HeLa cells.

Inonotus obliquus is a medicinal mushroom that has been used as an effective agent to treat various diseases such as diabetes, tuberculosis and cancer. Inotodiol, an included triterpenoid shows significant anti-tumor effect. However, the mechanisms have not been well documented. In this study, we aimed to explore the effect of inotodiol on proliferation and apoptosis in human cervical cancer HeLa cells and investigated the underlying molecular mechanisms. HeLa cells were treated with different concentrations of inotodiol. The MTT assay was used to evaluate cell proliferating ability, flow cytometry (FCM) was employed for cell cycle analysis and cell apoptosis, while expression of cyclinE, p27, bcl-2 and bax was detected by immunocytochemistry. Proliferation of HeLa cells was inhibited by inotodiolin a dose-dependent manner at 24h (r=0.9999, p<0.01). A sub-G1 peak (apoptotic cells) of HeLa cells was detected after treatment and the apoptosis rate with the concentration and longer incubation time (r=1.0, p<0.01), while the percentage of cells in S phase and G2/M phase decreased significantly. Immunocytochemistry assay showed that the expression of cyclin E and bcl-2 in the treated cells significantly decreased, while the expression of p27 and bax obviously increased, compared with the control group (p<0.05). The results of our research indicate that inotodiol isolated from Inonotus obliquus inhibited the proliferation of HeLa cells and induced apoptosis in vitro. The mechanisms may be related to promoting apoptosis through increasing the expression of bax and cutting bcl-2 and affecting the cell cycle by down-regulation the expression of cyclin E and up-regulation of p27. The results further indicate the potential value of inotodiol for treatment of human cervical cancer.

Epidemiology and clinical peculiarities of norovirus and rotavirus infection in hospitalized young children with acute diarrhea in Taiwan, 2009.

BACKGROUND/PURPOSE: Acute diarrhea is one of the most common morbidities in pediatrics worldwide. We conducted a study to investigate the incidence of norovirus in young children hospitalized with acute diarrhea in Taiwan and its clinical peculiarity compared with rotavirus gastroenteritis. METHODS: Between January and December, 2009, patients younger than 5 years and admitted to hospital with acute diarrhea were randomly selected; and their stool samples were collected and tested for presence of rotavirus and norovirus by enzyme immunoassay and reverse transcription-polymerase chain reaction, respectively. The clinical manifestations and laboratory findings of the enrolled patients were analyzed. RESULTS: A total of 989 cases were enrolled with a mean age of 21.6 ± 13.7 months and a male proportion of 56.0%. Rotavirus and norovirus was detected in 20.2% and 14.6% of all patients, respectively. Genogroup II was the predominant strain of norovirus (80.6%). Children aged 6-36 months accounted for the majority of patients positive for rotavirus and norovirus (73.0% and 81.3%, respectively). The incidences of norovirus and rotavirus infection were higher during winter and early spring. Most patients with rotavirus and norovirus diarrhea experienced vomiting (74.9%vs. 74.8%, respectively) and fever (94.7%vs. 71.3%, respectively). CONCLUSION: Most young diarrheal patients presenting with vomiting were likely to have norovirus or rotavirus infection. Patients with norovirus diarrhea experienced an absence of, or low-grade fever and longer duration of vomiting compared with those positive for rotavirus infection. A family history of current gastroenteritis may suggest the possibility of norovirus infection.

Distribution and expression of phosphorylated histone H3 during porcine oocyte maturation.

Phosphorylation modification of core histones is correlated well with diverse chromatin-based cell activities. However, its distribution pattern and primary roles during mammalian oocyte meiosis are still in dispute. In this study, by performing immunofluorescence and Western blotting, spatial distribution and temporal expression of phosphorylated serine 10 or 28 on histone H3 during porcine oocyte meiotic maturation were examined and distinct subcellular distribution patterns between them were presented. Low expression of phosphorylated H3/ser10 was detected in germinal vesicle. Importantly, following gradual dephosphorylation from germinal vesicle (GV) to late germinal vesicle (L-GV) stage, a transient phosphorylation at the periphery of condensed chromatin was re-established at early germinal vesicle breakdown (E-GVBD) stage, and then the dramatically increased signals covered whole chromosomes from pre-metaphase I (Pre-MI) to metaphase II (MII). Similarly, hypophosphorylation of serine 28 on histone H3 was also monitored from GV to E-GVBD, indicating dephosphorylation of histone H3 maybe involved in the regulation of meiotic resumption. Moreover, the rim staining on the chromosomes and high levels of H3/ser28 phosphorylation were observed in Pre-MI, MI, and MII stage oocytes. Based on above results, such stage-dependent dynamics of phosphorylation of H3/ser 10 and 28 may play specific roles during mammalian oocyte maturation.

Efficient generation of transgenic mice by direct intraovarian injection of plasmid DNA.

We established a rapid procedure for obtaining transgenic mice by directly injecting an enhanced green fluorescent protein (EGFP)-expressing plasmid (pIRES-EGFP) into the ovaries of fertile mice. The frequency of transgenic mouse production was determined by pair-mating, and by polymerase chain reaction (PCR) and sequence analysis of DNA taken from the tails of the offspring. The mice that received the EGFP gene transmitted it to their offspring (F(1)). Genetic and PCR analyses of F(1) progeny confirmed that the inserted EGFP was stably inherited. Of six female F(1) mice, all were able to pass the foreign DNA on to the next generation (F(2)). In situ hybridization using paraffin-embedded sections of ovarian and testicular tissues from the F(1) and F(2) progeny showed that the introduced gene was expressed in the gonads of the animals. The chromosomal location of the injected DNA was determined by fluorescence in situ hybridization, and the frequency of multiple site versus single site insertions is 85.71% (18/21) analyzed by FISH. We anticipate great progress in murine genetic engineering using this technique.