To explore the individual roles of G protein β and γsubunit in the interactions with effectors. Methods We investigated the interactions of Gβ1 and Gβ1γ2 with adenylyl cyclase Ⅱ(ACⅡ) us ing the yeast two-hybrid and three-hybrid systems. Results When assayed for the abilities to activate the reporter gene, the interactions among AD-β1, γ2 and BD-AC ⅡQ in the three-hybrid system were more po tent than the interactions between AD-β1 and BD-AC ⅡQ in the two-hybrid system. The expressions of BD-AC ⅡQ and AD-β1 in transformants coexpressed AD-β1 and BD-AC ⅡQ, and transformants coexpressed AD-β1, γ2 and BD-AC ⅡQ were respectively detected. The comparisons between the reporter activity and the expression levels of BD-AC ⅡQ and AD-β1 in the yeast cells show there was no correlations, i.e.The difference in the reporter activity was not a reflection of differential expression level of the hybrid pro teins. Conclutions All these results suggest that G protein β1 subunit is sufficient to maintain the basic in teraction between Gβ1γ2 and AC ⅡQ, and γ2 subunit plays an important role in the high affinity interaction of Gβ1γ2 with ACⅡQ.

One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.
Dianthus ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry

Objective:To evaluate the accuracy and influence factors of a triple-united method used for the determination of root canal working length.Methods:Root canal working length was detemined by manual measurement,tooth average length and radiographic measurement(triple-united method).In vitro experiment was conducted in 40 teeth and clinical study in 96 teeth by experienced specialists and postgraduates.Results:The accuracy of triple-united method was 82.5% and 62.5% by spcialist and poststudents in vitro, 87.1% and 75.5% in clinic,respectively. The accuracy was interfered by experience of the examiner, tooth integrity and apical disease.Conclusion:Triple-united method can meet the clinic requirement in the determination of root canal length.

In order to meet the requirements of cultivating the practical abilities and creativities of students who receive higher education, we initiated the reformation of education in the microbiology experiment teaching methods, implementing a system for module-based education, carefully monitoring every link in teaching, combining the encouragement and strict requirements together, adopting a proper way of assessment. It is proven that the implementation of the educational reformation mobilizes the interests of students and enhances the comprehensive qualities of students, which accomplishes the purposes of teaching.

Objective Lipid metabolism disorder can lead to male sterility, but its mechanism of affecting spermatogenesis and testis microenvironment remains unclear.This study aimed to investigate the effects of high fat diet on reproductive function and the expression of Ox-LDL within testis in male mice. Methods 16 C57BL/6 mice aged 8 weeks were divided into two groups by random number table method.High fat diet group was fed with high fat diet while normal fat diet group was fed with normal fat diet .At the end of 16 weeks, the levels of TG, TC, Ox-LDL and testosterone in serum were measured.The sperm concentration and motility from caudal epi-didymis were analyzed.The testis structure was observed by HE stai-ning.The localization and expression of Ox-LDL in testis were detec-ted by immunohistochemical technique. Results Compared with the normal diet group mice, the body weight (P<0.05) and the ser-um levels of TC [(2.31 ±0.33)mmol/L vs (6.54 ±0.31)mmol/L, P<0.01], Ox-LDL [(0.32 ±0.03)mg/L vs ( 0.44 ±0.06)mg/L, P<0.01] increased in high fat diet group mice significantly.The serum level of TG was of no significant difference(P>0.05).The serum level of testosterone[(3.64 ±0.43)mg/L vs (0.40 ±0.14) mg/L, P<0.01], the sperm concentration[(9.95 ±0.75)106/mL vs (5.66 ±1.51)106/mL, P<0.01] and the sperm motility[(54.69 ±17.84)%vs (32.48 ±5.80)%, P<0.01] decreased in high fat diet group significantly.HE staining also showed that the amount of Leydig cells, spermatids and spermatozoons reduced obvi-ously in high fat diet group.Immunohistochemistry staining showed that Ox-LDL was mainly distributed around Leydig cells of mice tes-tis.The expression of Ox-LDL in high fat diet group increased significantly ( P<0.01) . Conclusion The expression of Ox-LDL in Leydig cells of high fat diet C57BL/6J mice increases significantly, which may inhibit testosterone biosynthesis and affect spermatogen-ic function.

Objective To explore the protein factors that could interact with Gβ subunit within the G protein sig nal transducing pathway. Methods The highly sensitive protein-protein interaction system——Yeast Two-hybrid System was applied to screen the human cDNA library with constructed “Bite plasmid” contain ing Gβ subunit gene fragment. And then the faulse positive test was adapted. Results Three positive gene fragments were obtained. One codes for “Actin bundling protein”. The other two are new ones and their GeneBank accession numbers are AF288405 and AF288406 respectively. Conclusions It is the first time to find that among human brain tissue, Gβ subunit becomes a structural or functional unit interacts with actin bundling protein and the other two unknown protein factors to activate the following pathway. This re sult may be important to understand the relationship between the actin cytoskeleton and G proteins.

OBJECTIVEThough myasthenia gravis (MG) is a typical autoimmune disorder, there was some controversy on the treatment of the childhood-onset MG. By observing the efficacy of different therapies, the authors analyzed the affecting factors of prognosis in childhood-onset MG.

METHODThe retrospective data of 155 patients with childhood-onset MG (age of MG onset was less than 15 years) were collected from Department of Neurology, Beijing Tongren Hospital (January 2000 - February 2010). The patients were non-randomly divided according to their treatment into 3 groups (glucocorticoid, thymectomy and glucocorticoid combined with thymectomy groups). Postintervention status meeting the criteria of Myasthenia Gravis Foundation of America (MGFA) "complete stable remission, CSR", "pharmacologic remission, PR", "minimal manifestations, MM", or "Improved, I" was regarded as desirable response, which was used as primary indicator of observation. The authors assessed the efficacy of three therapies and analyzed the influencing factors of prognosis by using Chi-square test and Logistic regression.

RESULTAt 3 months of treatment, glucocorticoid group showed the highest effective rate. At the end of 1 year or 2 years of treatment, glucocorticoid combined with thymectomy group showed the highest effective rate respectively. The generalization rate of MG at 2 years, 10 years and 20 years in childhood-onset ocular MG patients were 4.3%, 10.7%, and 41.5%, respectively. Of patients with generalization of MG, 48.1% occurred within 2 years, 92.6% within 20 years. Univariate analysis showed that in childhood-onset ocular MG patients, variables such as age at onset (> 10 years), LG-MG and with chronic fatigue were significantly associated with general MG conversion. Whereas multivariate analysis showed that patients with age at onset (> 10 years) and chronic muscle fatigue were apt to convert to generalized MG.

CONCLUSIONGlucocorticoid appeared to have an effect that leads to early remission of symptoms in childhood-onset MG patients and glucocorticoid combined with thymectomy appeared to have better long-term effect. For those childhood-onset ocular MG patients with longer course of disease, older age of onset, chronic fatigue, or LG-MG, physicians should try to prevent the generalization of MG by immunosuppressive therapies.

Adolescent ; Child ; Child, Preschool ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Myasthenia Gravis ; diagnosis ; therapy ; Prognosis ; Retrospective Studies ; Thymectomy ; Young Adult

OBJECTIVETo explore the clinical characteristics of Henoch-Schönlein purpura (HSP) in children.

METHODSThe clinical data of 325 hospitalized children who were diagnosed with HSP between June 2012 and June 2014 were analyzed retrospectively.

RESULTSIn the 325 children with HSP, the incidence of HSP was higher in winter and spring, with 33.9% and 27.4%, respectively. Infection was the major factor to induce HSP (57.2%). The incidence of renal damage in children with purpura accompanied by abdominal symptoms and children with purpura accompanied by abdominal and joint symptoms was 60.3% and 48.9%, respectively, with statistically significant differences compared with children with purpura alone (P<0.05). In 32 children with purpura nephritis, the pathological grades of IIIa and IIIb were more common, accounting for 28% and 31%, respectively. In 325 children, an increased serum D-dimer level was observed in 260 children (80.0%), an increased peripheral IgA content in 101 children (46.3%), and a decreased CD4+ cell percentage in 62 children (56.4%).

CONCLUSIONSA high incidence of HSP is often seen in spring and winter. HSP is often induced by upper respiratory tract infection. Renal damage is more likely to occur in children with digestive tract symptoms, with IIIa and IIIb as the common pathological grades of renal damage.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Incidence ; Infant ; Kidney ; pathology ; Male ; Purpura, Schoenlein-Henoch ; complications ; epidemiology ; pathology ; Retrospective Studies ; Seasons

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

OBJECTIVETo elucidate the promoting effect of the constituents in plasma after oral administration of Liuwei Dihuangwan on proliferation of rat osteoblast.

METHODTo add the constituents in plasma after oral administration of Liuwei Dihuangwan with rational concentration into culture solution of rat osteoblast, later using MT method for determining the proliferation rate of rat osteoblast.

RESULTThe Mixed group including morroniside, sweroside and loganin with different dose all significantly promoted proliferation of rat osteoblast.

CONCLUSIONThis study preliminarily elucidated the constituents such as morroniside, sweroside and loganin in plasma after oral administration of Liuwei Dihuangwan were main biological constituents which contributed to anti-osteoporosis bioactivity of Liuwei Dihuangwan.

Administration, Oral ; Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Glucosides ; blood ; Glycosides ; blood ; Iridoid Glucosides ; Iridoids ; blood ; Osteoblasts ; cytology ; drug effects ; Plasma ; chemistry ; drug effects ; Rats ; Rats, Wistar