Sucking mice, 2～3 day after birth, were intraperiton eally inoculated with 0.05mL F1M10 of Chen Strain Hantaan virus. At diff erent t ime point after inoculation, the brains were taken, routinely fixed and embedded in paraffin for preparing 5 μm serial sections. Traditional and confocal immun ochemical detection of viral antigens and heat shock protein 70 were performed t o exp lore the cerebral stress response after viral infection. The results showed that HSP70 immunoreactivities could be stably detected in the viral antigen positive neurons, but not in the viral antigen negative or uninfected control tissues. B y confocal microscopic examination, the HSP70 and viral antigens were colocolize d in the neuronal cytoplasm. Our result, comparable to our previous findings in human tissues and culture cells, indicated that Hantavirus infection can induce the expression of HSP70 in the infected cells, and HSP70 expression might be n ecessary but not sufficient to keep the cell survival.

BACKGROUND: Previous animal experiments have shown that epimedium total flavonoids can exhibit preventive effects on estrogen-related bone loss, but few data are available in clinical research.OBJECTIVE: To investigate the effects of pimedium total flavonoids on bone mineral density (BMD) inprimary osteoporosis.DESIGN, TIME AND SETTING: A randomized, double-blinded, positively controlled clinical observation was performed at the Department of Traditional Chinese Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between June 2005 and September 2007.PARTICIPANTS: A total of 64 patients with primary osteoporosis consisting of 11 males and 53 females were included in this study. METHODS: All patients were randomly and evenly assigned into a treatment group and a control group. The treatment group was orally administered epimedium total flavonoids, 0.45 g once, three times daily, for a total of 6 months. Simultaneously, the control group was orally administered Gushukang, 10 g once, twice daily, for a total of 6 months. MAIN OUTCOME MEASURES: BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, greater trochanter, and left hip was measured, and simultaneously serum levels of calcium, phosphorus, and alkaline phosphatase (ALP) were detected pdor to and after treatment in each group.RESULTS: All 64 patients with primary osteoporosis were included In the final analysis, The BMD in the lumbar vertebrae (L1-4) was significantly higher in the treatment group than in the control group (P<0.05) and there was no significant difference In BMD in other sites between the two groups (P>0.05). Compared with pdor to drug application, BMD in the treatment group did not present obvious change after epimedium total flavonoids application, while in the control group, BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, and greater trochanter was significantly decreased (P<0.01-0.05). After drug application, serum level of calcium was significantly increased in each group, compared with prior to drug application (P<0.05), and no significant difference existed between the treatment and control groups (P>0.05). Tn the treatment group, serum levels of phosphorus and ALP did not alter significantly compared with prior to epimedium total flavonoids application (P>0.05).CONCLUSION: Epimedium total flavonoids exhibit effective effects on the maintenance of BMD in primary osteoporosis.

Objective To develop criteria for trainers of general practitioners in clinical training bases.Methods A primary version of criteria was established through literature review and expert interview.Thirty-two experts with middle-level professional title or above,who had 10 years of working experience or more in general practice,teaching and administration,were invited for 3 rounds of Delphi consultation during March and October 2013.Results The criteria consisted of 3 first-grade indicators and 12 second-grade indicators were established.The first-grade indicators included professional quality,clinical competence and teaching capability.The weight coefficient of first-grade indicators were 0.332 0,0.336 0 and 0.332 0,respectively.For 3 rounds of consultation the activity coefficient of experts was all 100％ ; the authority coefficients of experts were 0.881,0.897 and 0.883,respectively; and the harmonious coefficient of importance of the evaluation were 0.136,0.127 and 0.204,respectively (P ＜ 0.01).Conclusion The established criteria are credible and important for the selection of trainers of general practitioners in clinical bases,which would improve the quality of standardized training for general practitioners.

Objective To establish a set of accreditation standards for trainers of general practitioner (GP) in community health service centers.Methods A modified Delphi expert consultation was conducted during December 2012 and September 2013.Thirty two experts of general practice from different teaching hospitals and community health service centers were invited for three rounds Delphi questionnaires.A set of accreditation standards for GP trainers in primary care was established through data analysis,synthesis and process.Results Expert activity coefficients were 100.0％ for three rounds consultation,the authority coefficients were 0.894,0.882 and 0.893,respectively.The opinion coordination coefficients of experts were 0.156,0.166 and 0.215,respectively (P =0.000).The developed accreditation standards system comprised of three first-grade indicators and 14 second-grade indicators.The weight coefficients of three first-grade indicators,namely professional profiles,clinical competences in primary care and teaching abilities,were 0.339 8,0.335 6 and 0.324 6,respectively.Conclusions The established accreditation standard system is credible,which would appropriately guide the selection of GP trainers in primary care from three dimensions and enhance the quality of standardized training of general practitioners.

Objective To compare the efficacy of the reforming endoscopic minimally invasive releasing(REMIR) with open procedure for treatment of carpal tunnel syndrome.Methods Senventy patients with unilateral carpal tunnel syndrome were randomly divided into REMIR group and open procedure group,with 35 cases in each group.Kelly's standard,two-point discrimination,operation time and complication occurrence were compared between the two groups.Results All patients were followed-up for 12 months.There was no significant difference in the therapeutic results and in the improvement of two-point discrimination between the two groups (P ＞ 0.05 ).The operation time of REMIR group was significantly shorter than the open procedure group ([ 10.03 ± 1.84] min vs [37.63 ±7.18]min,t =22.210,P ＜0.001 ).And there was no cases with scar tenderness in REMIR group while there was 7 cases in open procedure group.Conclusion Compared with the open procedure,the REMIR method has the same clinical efficacy while with the advantages of causing smaller skin scar and being less time consuming.

BACKGROUNDThe Toll-like receptors (TLRs) represent a group of single-pass transmembrane receptors expressed on sentinel cells that are central to innate immune responses.The aim of this study was to investigate the presence of soluble TLRs in pleural effusions, and the diagnostic values of TLRs for pleural effusion with various etiologies.

METHODSPleural effusion and serum samples were collected from 102 patients (36 with malignant pleural effusion, 36 with tuberculous pleural effusion, 18 with bacterial pleural effusion, and 12 with transudative pleural effusion). The concentrations of TLR1 to TLR10 were determined in effusion and serum samples by enzyme linked immunosorbent assay. Four classical parameters (protein, lactate dehydrogenase, glucose and C-reactive protein (CRP)) in the pleural fluid were also assessed. Receiver-operating characteristic curves were used to assess the sensitivity and specificity of pleural fluid TLRs and biochemical parameters for differentiating bacterial pleural effusion.

RESULTSThe concentrations of TLR1, TLR3, TLR4, TLR7 and TLR9 in bacterial pleural effusion were significantly higher than those in malignant, tuberculous, and transudative groups, respectively. Analysis of receiver operating characteristic curves revealed that the area under the curves of TLR1, TLR3, TLR4, TLR7 and TLR9 were 0.831, 0.843, 0.842, 0.883 and 0.786, respectively, suggesting that these TLRs play a role in the diagnosis of bacterial pleural effusion. Also, the diagnostic value of TLRs for bacterial pleural effusions was much better than that of biochemical parameters (protein, lactate dehydrogenase, glucose and CRP).

CONCLUSIONSThe concentrations of TLR1, TLR3, TLR4, TLR7 and TLR9 appeared to be increased in bacterial pleural effusion compared to non-bacterial pleural effusions. Determination of these pleural TLRs may improve the ability of clinicians to differentiate pleural effusion patients of bacterial origin from those with other etiologies.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacterial Infections ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Pleural Effusion ; metabolism ; microbiology ; Prospective Studies ; Toll-Like Receptor 1 ; metabolism ; Toll-Like Receptor 3 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Toll-Like Receptor 7 ; metabolism ; Toll-Like Receptor 9 ; metabolism ; Toll-Like Receptors ; metabolism ; Young Adult

OBJECTIVETo study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma.

METHODSA total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA).

CONCLUSIONSGlucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glucose ; physiology ; Glucose Transport Proteins, Facilitative ; genetics ; metabolism ; Glucose Transporter Type 1 ; genetics ; metabolism ; Humans ; Prognosis

OBJECTIVETo objectively evaluate the clinical effect of traditional Chinese medicine in treating children's respiratory syncytial viral pneumonia (RSVP) of phlegm-heat blocking Fei syndrome (PHBFS).

METHODSA single-blinded multi-center, blocked, randomized and parallel-controlled method was adopted. The clinical study was carried out on 206 children with RSVP-PHBFS who were assigned to two groups, 108 in the test group treated through intravenous dripping of Qingkailing Injection () in combination of oral intake of Er'tong Qingfei Oral Liquid () and 98 in the control group with intravenous dripping of ribavirin injection in combination with oral intake of potassium guaiacol sulfonate oral liquid, all for 10 days. The clinical efficacy was evaluated and compared at the end of the trial from various aspects by three methods including comprehensive efficacy, post-treatment main symptoms score difference and survival analysis of the main symptoms.

RESULTSAfter treatment, in the test group, 60 patients were cured, 36 markedly alleviated, and 12 improved. In the control group, 41 were cured, 38 markedly alleviated, 18 improved and 1 unchanged. Comparison on the comprehensive efficacy between the two groups shows a better efficacy in the test group (chi(2)=4.4527, P=0.0348). Scores of the main symptoms were lowered after treatment in both groups, the difference was 22.41+/-4.99 scores in the test group and 17.61+/-6.34 scores in the control group, being more significant in the former (t=-5.99, P<0.01). Survival analysis shows that there was significant difference between the two groups in the effect initiating time on such symptoms as fever, cough, copious sputum, shortness of breath, and rales, which was earlier in the test group (P<0.01 or P<0.05).

CONCLUSIONEvaluation of the efficacy of traditional Chinese medicine in treating children with RSVP-PHBFS by using the three methods jointly could better show the objectivity of the evaluation.

Airway Obstruction ; complications ; mortality ; therapy ; Child, Preschool ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fever ; complications ; mortality ; therapy ; Hot Temperature ; Humans ; Infant ; Male ; Medicine, Chinese Traditional ; methods ; Pneumonia, Viral ; drug therapy ; mortality ; pathology ; Respiratory Syncytial Virus Infections ; drug therapy ; mortality ; pathology ; Respiratory System ; pathology ; Ribavirin ; administration & dosage ; Single-Blind Method ; Survival Analysis ; Syndrome ; Treatment Outcome

BACKGROUNDPrenatal hyperglycaemia may increase metabolic syndrome susceptibility of the offspring. An underlying component of the development of these morbidities is hepatic gluconeogenic molecular dysfunction. We hypothesized that maternal hyperglycaemia will influence her offsprings hepatic peroxisome proliferator-activated receptor coactivator-1α (PGC-1α) expression, a key regulator of glucose production in hepatocytes.

METHODWe established maternal hyperglycaemia by streptozotocin injection to induce the maternal hyperglycaemic Wistar rat model. Offspring from the severe hyperglycemia group (SDO) and control group (CO) were monitored until 180 days after birth. Blood pressure, lipid metabolism indicators and insulin resistance (IR) were measured. Hepatic PGC-1α expression was analyzed by reverse transcription polymerase chain reaction and Western blotting. mRNA expression of two key enzymes in gluconeogenesis, glucose-6-phosphatase (G-6-Pase) and phosphoenolpyruvate carboxykinase (PEPCK), were analyzed and compared.

RESULTSIn the SDO group, PGC-1α expression at protein and mRNA levels were increased, so were expression of G-6-Pase and PEPCK (P < 0.05). The above effects were seen prior to the onset of IR.

CONCLUSIONThe hepatic gluconeogenic molecular dysfunction may contribute to the metabolic morbidities experienced by this population.

Animals ; Female ; Hyperglycemia ; chemically induced ; physiopathology ; Insulin Resistance ; physiology ; Liver ; metabolism ; Male ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; RNA-Binding Proteins ; Rats ; Rats, Wistar ; Streptozocin ; toxicity ; Transcription Factors

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
Citrullus ; genetics ; Fruit ; genetics ; Gene Expression Regulation, Plant ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Sequences, Nucleic Acid ; genetics ; physiology