We wished to select a cold-adapted genotype G1P[8] ZTR-68 rotavirus (China southwest strain) in MA104 cells for possible use as a live vaccine. ZTR-68 was recovered originally from children with diarrhea. The virus was cultivated at 37 degrees C at the first passage. Then, the cultivation temperature was decreased stepwise by 3 degrees C per eight passages. In total, the virus was passaged 32 times, and cultivation was terminated at 28 degrees C. Biological characteristics of the virus were analyzed during serial passages. There was no difference between the migration patterns of genomic dsRNA segments according to polyacrylamide gel electrophoresis of original and cold-adapted viruses. Infectious and red cell-agglutination titers of cold-adapted virus were lower than those of the parent virus. Also, the virus formed small-size plaques with irregular shapes at 31 degrees C and 28 degrees C. These results suggested that a genetically stable attenuated virus can be obtained through serial cold-adapted passages. Thus, an alternative strategy is provided by cold-adaption for development of attenuated live rotavirus vaccines.
Adaptation, Physiological ; China ; Cold Temperature ; Diarrhea ; virology ; Female ; Genotype ; Humans ; Infant ; Male ; Rotavirus ; genetics ; growth & development ; isolation & purification ; physiology ; Serial Passage ; Virus Cultivation ; Virus Replication

Eleven C21 steroids were isolated from chloroform extract of roots of Cynanchum otophyllumby silica gel, MCI, ODS columns, and semi-preparative HPLC. Their structures were determined by spectroscopic data analysis as otophylloside B(1), caudatin-3-O-beta-D-cymaropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (2), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (3), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-digitoxopyranosyl-(1-->4)-beta-D-cymaropyranoside (4), otophylloside O (5), gagamine-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (6), sinomarinoside B (7), mucronatosides C (8), wallicoside J (9), stephanoside H (10), and qinyangshengenin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-digitoxopyranoside (11). Among them, compounds 2-3, and 6-11 were separated from the roots of this plant for the first time.
Cynanchum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots ; chemistry ; Steroids ; chemistry ; isolation & purification

Objective:To investigate the application effects of SBAR communication mode (situation, background, assessment and recommendation) combined with 3D printing model technology in bed-side teaching of trauma orthopaedics.Methods:A total of 80 clinical medicine students were randomly divided into two groups according to the order of the school number, with 40 students in each group. One was experimental group which received SBAR communication mode combined with 3D printing model technology teaching, and the other one was the control group, which received regular reaching mode. At the end of teaching, the teaching effects were evaluated, including the scores of theoretical examinations and operational examinations, total scores and the anonymous questionnaires. SPSS 22.0 was used for recording and statistical analysis.Results:The average scores of theoretical examinations (48.30±1.41), operational examinations (42.20±1.48) and total scores (90.70±1.38) of experimental group were significantly higher than those of control group [(43.40±1.52); (34.80±1.53); (78.10±1.51)], with significant differences ( P <0.05). The anonymous questionnaires showed that the students in the experimental group had a significant advantage in autonomous learning ability, learning enthusiasm, the ability of literature retrieval and analysis and clinical thinking ability, and enhancing students' humanistic care consciousness in clinical work ( P <0.05). While the two groups had the same recognition in improving problem solving ability, teamwork ability and communication ability with patients, with no significant difference ( P > 0.05). Conclusion:The new teaching mode, SBAR mode combined with 3D printing model technology, applied to trauma orthopaedics bed-side teaching is helpful for students to improve their learning interest and autonomous learning ability, cultivate their lifelong learning habits and their comprehensive quality, so this mode will significantly improve the teaching effects, with good application value.

Objective:To evaluate the effect of remote ischemic conditioning (RIC) on left ventricular (LV) myocardial perfusion, myocardial viability, LV remodeling, regional and global LV function serially following acute myocardial infarction (AMI) in Chinese mini-pigs.Methods:AMI was established in 12 Chinese mini-pigs (8 males, 4 females; age: 6-8 months) and they were randomly divided into RIC group ( n=6) and non-RIC group ( n=6). RIC was performed in pigs by blood pressure inflation on the lower limbs for 5 min period and 4 cycles immediately after surgery. A series of myocardial perfusion imaging and gated 18F-fluorodeoxyglucose (FDG) myocardial metabolism PET/CT imaging were performed longitudinally at the 1st, 14th, 28th and 56th days after AMI, and parameters including total perfusion defect (TPD), hibernating myocardium (HM), Scar, left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), summed motion score (SMS), summed thickening score (STS) and changes of these parameters were obtained. Independent-samples t test and Mann-Whitney U test were used to analyze data. Results:Nine Chinese mini-pigs survived after surgery and were performed imaging. Compared to non-RIC group ( n=4), HM at the 28th ((6.0±2.4)% vs (17.0±4.6)%; t=-4.158), TPD 14th-1st ((-11.8±5.4)% vs 9.0%(4.5%, 15.0%); z=2.449), TPD 28th-1st ((-15.3±3.9)% vs (12.0±3.0)%; t=-10.071), TPD 56th-1st ((-18.0±6.5)% vs 9.0%(4.5%, 12.0%); z=2.449), HM 28th-1st ((-10.5±6.9)% vs (8.3±2.1)%; t=-4.507), HM 56th-1st (-15.0%(-17.5%, -8.5%) vs 2.0%(0%, 7.0%); z=2.449) and LVEDV 14th-1st (-0.5(-2.5, 0) ml vs (13.0±4.4) ml; z=2.470) were reduced in RIC group ( n=5; all P<0.05). Conclusion:RIC can improve myocardial perfusion, delay LV remodeling in the acute stage and salvage hibernating myocardium in the subacute stage and chronic stage.

Objective To investigate the advantage and application prospect of 3D printing technology in assisting surgery for complex tibial plateau fractures.Methods The complete clinical data of 41 patients (48 knees) were retrospectively analyzed.And these patients were divided into two groups as 3D printing assisted operation group and traditional operation group,according to whether 3D printing technology was applied.Study was carried out to compare the clinical efficacy of surgery assisted by 3D printing technology and traditional surgery in the treatment of complex tibial plateau fractures.The 3D printing assisted operation group included 18 patients (22 knees,12 males and 6 females),aging from 16 to 68 (mean age 45.5±7.2),and there were 12 cases of Schatzker Ⅴ and 10 Ⅵ.The traditional operation group included 23 patients (26 knees,15 males and 8 females),aging from 19 to 69 (mean age,46.2±6.8),and there were 14 cases of Schatzker Ⅴ and 12 Ⅵ.The operation time,intraoperative blood loss,the Rasmussen score and hospital for special surgery knee score (HSS) at 6 weeks post-operation were analyzed and the difference between the two groups was tested.Results All those patients were followed up for 7 to 20 months (mean 15 months).For 3D printing assisted operation group,the mean operation time was 81.4±6.3 min for Schatzker Ⅴ and 90.6± 15.4 min for Schatzker Ⅵ;the mean intraoperative blood loss was 200.4±72.3 ml for Schatzker Ⅴ and 280.6±101.6 ml for Schatzker Ⅵ;the result of 6-week Rasmussen score was 14 excellent,6 good and 2 fair (excellent and good rate 90.9％);the result of 6-week HSS score was 15 excellent,5 good and 2 fair (excellent and good rate 90.9％).Then for traditional operation group,the mean operation time was 100.4± 15.3 min for Schatzker Ⅴ and 111.5±20.2 min for Schatzker Ⅵ;the mean intraoperative blood loss was 450.4±173.3 ml for Schatzker Ⅴ and 500.5±247.2 ml for Schatzker Ⅵ;the result of 6-week Rasmussen score was 14 excellent,8 good,2 fair and 2 bad (excellent and good rate 84.6％);the result of 6-week HSS score was 13 excellent,8 good,2 fair and 3 bad (excellent and good rate 80.8％).The difference in operation time,intraoperative blood loss had statistical significance between the two groups;but the difference in 6-month post-operation Rasmussen score and HSS score had no statistical significance.Conclusion Compared with traditional surgery,3D printing technology assisted surgery for complex tibial plateau fracture possesses advantage such as shortened operation time,reduced intraoperative blood loss;but there's no evidence for improved knee function at 6 weeks post-operation.

To verify the effect of echinacoside on replication and antigen expression of hepatitis B virus (HBV) by using HBV-transfected HepG2. 2. 15 cells as the in vitro model. The ELISA method was used to determine HBeAg and HBsAg levels in cellular supernatants. The effect of echinacoside on HBV replication was studied by using HBV transgenic mice as the in vivo model. First of all, the HBV DNA level in hepatic tissues was quantified with PCR method. Meanwhile, the serum transaminase levels and hepatic pathological changes were also evaluated. Subsequently, HBV transgenic mice were divided into five groups: the control group, the lamivudine group (50 mg · kg(-1)) and echinacoside high, medium and low dose group (50, 25 and 12.5 mg · kg(-1)). The mice were orally administered with drugs once per day for 30 days. At the 31st day, the mice serum was separated to measure HBsAg, HBeAg and HBV DNA. Additionally, the liver HBV DNA level and histopathological change were detected. The results indicated that echinacoside at 50 and 100 mg · L(-1) suppressed significantly HBsAg and HBeAg expressions on the sixth day, with the maximum inhibition ratios of 42.68% and 46.29%; And echinacoside at 100 mg · L(-1) also showed an inhibitory effect on HBV DNA. Besides, echinacoside at 50 mg · kg(-1) inhibited significantly HBsAg and HBeAg expressions of HBV transgenic mice, with the inhibition ratios of 42.82% and 29.12%, and reduced markedly the serum HBV DNA level in HBV transgenic mice. In conclusion, the study suggested that echinacoside has a strong effect against HBV replication and antigen expression.
Animals ; DNA, Viral ; blood ; Female ; Glycosides ; pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Virus Replication ; drug effects

OBJECTIVETo study the subcellular localization of severe fever with thrombocytopenia syndrome virus (SFTSV) in macrophages and understand the replication and assembly mechanism of SFTSV in host cells.

METHODSUsing two types of human macrophage cell lines THP-1 and U937, the study analyzed the intracellular colocalization of SFTSV with Golgi apparatus and endoplasmic reticulum by immunefluorescence staining and confocal microscopy.

RESULTSSFTSV infected macrophage cell lines THP-1 and U937. Immunofluorescence staining showed that the SFTSV nuclear protein colocalized with Golgi apparatus and closely surrounded by endoplasmic reticulum in the perinuclear region.

CONCLUSIONThe results suggested that Golgi complex and endoplasmic reticulum are probably the sites for formation and maturation of SFTSV viral particles.

Bunyaviridae ; isolation & purification ; Cell Line, Tumor ; Endoplasmic Reticulum ; virology ; Fever ; virology ; Golgi Apparatus ; virology ; Humans ; Macrophages ; virology ; Thrombocytopenia ; virology

OBJECTIVETo explore the relative factors on the failure in digit replantation in order to take preventions to control the risk factors.

METHODSFrom January 2013 to December 2013, 236 consecutive patients (311 fingers) underwent digit replantation were collected to analyze retrospectively, involving 183 males and 53 females with an average age of 34.5 years old ranging from 2 to 62 years old (6 cases under 6 years old and 230 cases elder than 6 years old). There were 51 thumbs, 87 index fingers, 78 middle fingers, 63 ring fings and 32 little thumbs. Forty cases(forty fings) who were failured as the observation group, the others as the control group. The factors of age, gender, finger, cause of injury, smoking history, ischemia duration, plane of division, condition of venous drainage and condition of arterial repair we assessed.

RESULTSAll 236 cases with 311 fingers were replanted, 40 fingers were failured after operation. The relative factors on the failure in digit replantation included smoking history, cause of injury, plane of division, condition of venous drainage and condition of arterial repair (P< 0.05). There were no significant correlation between the failure and age, gender, finger and ischemia duration (P>0.05).

CONCLUSIONSmoking history, causes of injury, plane of division, condition of venous drainage and condition of arterial repair are risks of failure in digit replantation. Before choosing the type of operation, it should be think about the patient's general conditions, injury status, grasp firmly the operative indications and actively carry out surgical treatment.

Adolescent ; Adult ; Child ; Female ; Finger Injuries ; surgery ; Fingers ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Replantation ; Retrospective Studies ; Risk Factors ; Thumb ; injuries ; surgery ; Treatment Failure ; Young Adult

Objective To find out the relationship of ecological environment protection and the transmission of Lyme disease under economic development of western regions in China.Methods Both scene molecular and traditional epidemiological methods were used to assess the effects of environmental protection on the transmission of Lyme disease.Results Among areas as protected natural forests,semiprotected nursery forests and farmland,the vector tick species and reservoir rodents from protected natural forests area had the highest quantity of population and diversity index and followed by semi-protected nursery forests.Vector competence of reservoir hosts and value of natural foci from protected natural forests area were also remarkably higher than those areas of semi-protected nursery forests and farmland. Staff working in the areas who were bitten by ticks from protected natural forests areas had higher serological Dositive rate(66.7%)than those from semi-protected nursery forests areas(2.5%),and both showed remarkable difference(P=6.45,E-11＜0.01,df=1).The difference of genetic divergence among these subpoPulations from different habitats being surveyed showed that the biggest genetic divergence index (Fst)of 0.557 42 was between protected natural forests area and farmland area.The index between semiprotected nursery forests area and farmland area was also bigger than zero with statistical significance.The genetic divergence index of 0.108 02 between semi-protected nursery forests area and protected natural forests area was the lowest which showed that genetic divergence between the subpopulations of the two sampling areas was not obvious.The genetic distance among these subpopulations had similar change along with their habitats.Conclusion Under economic development of western regions in China,when programs as natural forests protections,recovery prairie and grassland from farmland were actively performed,vectors insects and reservoir hosts of Lyme disease might also be protected to some degree but the risk and value of natural foci on Lyme disease might increase.Data suggested that people entering these areas should be told to strengthen their awareness on individual protection against the disease.

Objective:To observe the effect of point-moxibustion on gastrointestinal motility,mRNA and protein expressions of ghrelin and growth hormone secretagogue receptor 1a (GHSR-1a) in lateral septal nucleus of rats with diabetic gastroparesis (DGP),and to investigate the central regulatory mechanism of DGP treatment with point-moxibustion.Methods:Forty SPF-grade Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,an electroacupuncture (EA) group and a point-moxibustion group,with 10 rats in each group.A DGP rat model was established by intraperitoneal injection of 2％ streptozotocin (STZ) with 8-week irregular high-sugar and high-fat diet in the model group,the EA group and the point-moxibustion group;and rats in the blank group were injected intraperitoneally with 0.1 mmoL/L (pH 4.5) citric acid-sodium citrate buffer with 8-week normal diet.Eight weeks later,rats in the EA group were treated by EA at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6);while rats in the point-moxibustion group were treated by point-moxibustion at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6) for successive 15 d.Rats in the blank group and the model group were fixed as the control without intervention.After treatment,intestinal propulsion rate and gastric emptying rate were measured.The mRNA and protein expressions of ghrelin and GHSR-1a in the lateral septal nucleus were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB).Results:Compared with the blank group,the intestinal propulsion rate and gastric emptying rate of the model group were significantly lower (both P＜0.01);compared with the model group,the intestinal propulsion rate and gastric emptying rate of the EA group and the point-moxibustion group increased significantly (all P＜0.05).The mRNA and protein expressions of ghrelin and GHSR-1a were lower in the model group than those in the blank group (all P＜0.01).The mRNA and protein expressions of ghrelin and GHSR-1a were significantly higher in the EA group and the point-moxibustion group than those in the model group (all P＜0.05).There were no statistically significant differences between the EA group and the point-moxibustion group (all P＞0.05).Conclusion:Point-moxibustion at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6) can increase the intestinal propulsion rate and gastric emptying rate of DGP rats,and promote the mRNA and protein expressions of ghrelin and GHSR-1a in the central nervous system.The mechanism may be related to the activation of ghrelin pathway in hypothalamic arcuate nucleus-lateral septal nucleus.