Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Objective To evaluate the clinical efficacy of interventional occlusion of ventricular septal rupture (VSR) complicating acute myocardial infarction (AMI). Methods Six patients with VSR complicated after AMI underwent transcatheter interventional occlusion in Chinese PLA General Hospital between May 2009 to May 2015 were retrospectively analyzed. Their clinical features, interventional treatment protocols, and occurance of postoperative complications were studied. Results Among the 6 patients, VSR were successfully occluded in 5 patients. One patient failed the operation due to instability of occluder after it was deployed and the occluder was retrieved. The paitents died of heart failure 6 months later. Among the 5 patients with successful closure, 4 patients presented mild residual shunt after occlusion and acute left heart failure occurred in 1 patient after operation. Two patients died during hospital stay after operation. Between them, one patient died 3 hours after operation because of cardiac tamponade and the other patient died of cardiogenic shock after withdrawal from IABP. Three patients were followed up until now and follow up echocardiography showed satisfactory cardiac function without heart failure. Conclusions Transcatheter occlusion was a reliable therapy for patients with ventricular septal rupture complicated in acute myocardial infarction which could improve cardiac function and reduce mortality.

Adult ; Aortic Coarctation ; therapy ; Catheterization ; Humans ; Male ; Platinum ; Stents

Background Qiang minority is minority groups of China with the special habits and customs and living condition. So whether the spectrum of disease and bacteria spectrum in conjunctiva are similar with Han nationality is worth paying attention. Objective Present survey was to obtain the data about bacterial species in conjunctival sac in Qiang minority population with the age 40 years old and more and the compare with matched Han nationality population. Methods This survey study was performed as the standardized training and protocol. A total of 212 eyes of 106 individuals from Qiang minority in Beichuan county and 640 eyes of 320 subjects from Han nationality in Mianyang city received questionnaire survey and ophthalmological examination. The secretion of the inferior palpebral conjunctival sac was embrocated and inoculated on blood plate for 48-72 hours. The bacteria was separated and identified. This study was approved by the Ethic Committee of Sichuan Provicial People' s Hospital. Orally informed consent was obtained before the medical procedure. Results All the examinee finished the survey and examination with a good compliance. No significant difference was found in the demography between these two groups of population. The multiple bacterial positive rate in conjunctival sac was 59. 4% in Qiang minority and that of Han people was 66. 3% with a considerably difference between them (χ2 = 2. 27,P = 0. 13). The multiple bacterial species were simultaneously detected in 26.2% in Qiang minority population and 11.88% Han people, showing evidently difference (χ2 = 106. 40, P = 0. 00 ) . The positive rate of corynbaccterium in conjunctival sac of Qiang minority was statistically lower than that of Han people (20. 7% versus 45. 0% ,χ2 =31. 75 ,P = 0. 00) ,but there was no statistical difference in the positive rate of staphylococcus epidemics between two groups (χ2 = 1. 89 ,P = 0. 17). Conclusion The bacteria positive rate in conjunctiva sac is resemble in the population over 40 years in both the Qiang minority and Han nationality. The simple bacterial species is found in majority people in two groups of subjects. The positive rate of multiple bacterial strains coexistence is more in the Qiang minority. The bacterial strains is different between Qiang minority and Han nationality.

Background Epidemiological surveys showed that the types of bacteria are different in the conjunvical sac from different nationalities,which possibly is associated with living environment.To characterize the types of conjunctival bacteria involved is important for the prevention and treatment of infectious eye diseases.Objective The present survey was to obtain data about bacterial species in the conjunctival sac in the Tibetan minority population aged over 40 years old and compared with the matched Han nationality population.Methods The standardized training and protocol were performed before this survey.A total of 290 eyes of 145 individuals from the Tibetan minority and 346 eyes of 173 subjects from the Han nationality were enrolled in this study in Ganzi Autonomous Prefecture,who had received questionnaire surveys and ophthalmological examinations.The secretion of the inferior palpebral conjunctival sac was embrocated and inoculated and grown on blood plates for 48-72 hours.The bacteria were isolated and identified.This study was approved by the Medical Ethic Committee of the Sichuan People Hospital.Oral informed consent was obtained from the subjects.Results No significant differences were seen in the constituent ratio of the gender as well as the age between the Tibetan minority and Han nationality in this study (x2 =0.987,P=0.3202;t=1.142,P=0.254).There was a significant difference in the proportions of farmers and herdsmen between the two groups(x2 =8.557,P =0.000).The positive rate of bacterial cultivation in Tibetan individuals was 50.74％,showing a statistically significant decrease in comparison with the Han people(60.4％)(x2=6.042,P=0.014).There was no statistical difference in the multiple bacterial species between the two groups (11.0％ in Tibetan,11.6％ in Han people)(x2 =0.0271,P =0.869).The rate of staphylococcus epidemics was 26.6％ in the Tibetan minority and that of Han population was 33.2％,without a significant difference between them (x2 =3.350,P=0.060).No significant difference was seen in the ratio of corynbacterium infection between the two population(15.9％ vs.17.3％)(x2 =0.248,P =0.618).Conclusions The ratio of bacterial cultivation in Tibetans is statistically lower than that of the Han people.The types and distribution of bacteria are similar in the Tibetan and Han nationality.

OBJECTIVEThe study aims to find out the fluctuating curve of the intra-articular pressure in temporomandibular joint with sudden-onset, persistent, severe closed lock and discuss the mechanism of its formation. We also investigate the effects of the arthrocentesis.

METHODS20 affected sides in 16 patients were collected. A No. 8 syringe needle was used to pierce into the upper compartment of TMJ. The pre-arthrocentesis intra-articular pressure was measured by the pressure transducer via the flexural rigid tubing at open and close bite. The curve and mean value were documented during the operation.

RESULTSThe patients with sudden-onset, persistent, severe closed lock had significantly low negative intra-articular pressure in their affected temporomandible joints. The average pressures was (-9.947 +/- 8.854) kPa at open bite and (-6.475 +/- 4.147) kPa at close bite.

CONCLUSIONThe TMJs with sudden-onset, persistene, severe closed lock has particular characters on etiology and clinical behavior. Arthrocentesis is one of the effective treatments to the diseases.

Adult ; Dental Occlusion ; Female ; Humans ; Male ; Middle Aged ; Paracentesis ; Range of Motion, Articular ; Temporomandibular Joint ; Temporomandibular Joint Disorders ; Treatment Outcome

OBJECTIVETo observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.

METHODSOne hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.

RESULTSOA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).

CONCLUSIONS(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.

Animals ; Antlers ; chemistry ; Cartilage ; cytology ; metabolism ; Chondrocytes ; drug effects ; metabolism ; Female ; Medicine, Chinese Traditional ; Osteoarthritis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism

Objective To observe the clinical efficacy and safety of voriconazole tablets combined with caspofungin injection in the treatment of hematological disease with invasive pulmonary aspergillosis.Methods A total of 100 patients of hematologic disease with invasive pulmonary aspergillosis were randomly divided into control group and treatment group with 50 cases per group.Control group was given amphotericin B,the first dose was 5 mg,the maximum dose of 25 mg at a speed of 5 mg · d-1,and then the dose of 25 mg · d-1 for 10 days,qd,intravenous drip,which was treated for 15 d.Treatment group was given caspofungin,the first dose of 70 mg,adjusted to 50 mg,qd,intravenous drip + voriconazole,the first day of 6 mg · kg-1,the second day adjusted to 4 mg · kg-1,bid,orally,which was treated for 15-28 d (treatment to full effect).The clinical efficacy and adverse drug reactions were compared between two groups.Results After treatment,the main indexes in treatment and control groups were compared:the total effective rates were 88.00％ (44/50 cases) and 68.00％ (34/50 cases),the total effective rates with empirical treatment were 92.31％ (24/26 cases) and 81.48 ％ (22/27 cases),the total effective rates with proactive treatment were 83.33％ (20/24 cases) and 52.17％ (12/23 cases),the differences were statistically significant (all P ＜ 0.05).The adverse drug reactions in treatment group were based on nervous system damage and liver and kidney dysfunction,which in control group were based on nervous system damage,peripheral edema,liver and kidney dysfunction and visual abnormalities.The incidences of adverse drug reactions in treatment and control groups were 4.00％ and 14.00％ with significant difference (P ＜ 0.05).Conclusion Voriconazole tablet combined with caspofungin injection has definitive clinical efficacy and safety in the treatment of hematological disease with invasive pulmonary aspergillosiss.

Anticonvulsants ; adverse effects ; Child ; Child, Preschool ; Drug Hypersensitivity ; diagnosis ; etiology ; therapy ; Female ; Humans ; Infant ; Male

Human cytomegalovirus (HCMV) is extremely species specific and does not replicate in experimental animal tissues.To overcome the problem and establish suitable animal models for studying antiviral strategies,the expression of HCMV UL49 gene was explored in mice.UL49-GFP gene was subcloned into the adenovirus shuttle plasmid pDC316,the products(pDC316-UL49-GFP)were co-transfected with helper plasmid pBHGloxE1,3Cre into HEK293 cell lines by liposome reagent,recombinant adenovirus(Ad-UL49-GFP) was generated and confirmed by PCR and Western blot.Ad-UL49-GFP was propagated in 293 cells and purified.The titer of viral stocks was determined by end-point dilution assay.The purified adenoviruses were delivered into mice via the tail vein injection.Fluorescence quantitative PCR and Western blot experiments were used to examine the tissue distribution and duration of UL49 gene expression.The results showed that the recombinant adenovirus were present in vivo.The expression level in tissues arranged in descending order was liver,spleen,kidney,heart and lung.3 days after injection,the liver,spleen,kidney,heart and lung expressed protein UL49 in high lever and then declined gradually.14 days after injection,UL49 protein expression was disappear in some organs except liver and spleen.In conclusion,transgene animal model carrying UL49 gene was successfully established.Therefore,the system may be suitable for selecting anti-HCMV drugs targeting UL49 gene.