Objective To provide the initiative experimental reference for resource utilization of Cynanchum komarovii by biological technology. Methods Using root, stem, and leaf of C. komarovii as explants, the proper media for inducing of callus were optimized by orthogonal test. In culturing process for proliferation of callus, the fresh and dry weights were determined at various times and callus growth curve was drawn. Meanwhile, division test of callus was taken. Results Root callus test showed the best effect in successful inducing percentage of 90%-100%. The best media combination of stem and leaf were MS+2,4-D 1.0 mg/L+6-BA 2.0 mg/L+NAA 0.5 mg/L and MS+2,4-D 0.5 mg/L+6-BA 0.5 mg/L+NAA 0.5 mg/L. The stem segament was more easily induced than leaf. The callus induced by various treatments is in loose character and fresh yellow colour with no brown phenomenon. At day 10 the growth entered the exponent period of callus offspring, the maximal fresh weight to 4.961 g and the dry weight to 0.496 g. The division rate of callus was rather low and not easy to shoot. However, the nail root was commonly seen in the test. Conclusion The root is proved to be the best explants in the test of inducing callus, which shows the best medium of stem and leaf for forming the callus. The growth of callus is bigger, but the division is not easy.

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

ABSTRACT:Objective To evaluate the features and related factors of decremental response in amyotrophic lateral sclerosis (ALS)patients to low-frequency repetitive nerve stimulation (RNS)in proximal nerves.Methods We performed RNS studies in proximal axillary and accessory nerves with recording in deltoid and trapezius mus-cle respectively in 87 ALS patients.Decremental compound muscle action potential (CMAP)and related factors were analyzed prospectively,and abnormal group of decremental response in ALS patients was compared with 39 pa-tients with myasthenia gravis.Results ① Abnormal decremental responses were found in 43.7% and 49.4% of ALS patients with deltoid and trapezius muscle recording respectively.They were found more frequent in trapezius muscle than those of deltoid muscle.② There was no relationship of decremental response with gender,age,onset or course of disease,ALSFRS-r scores,or rate of disease progression in ALS patients.③ There was significant rela-tionship between decremental response and fluctuating muscle weakness.Decremental responses decreased more ob-viously in ALS patients with fluctuating muscle weakness than in those with nonfluctuating muscle weakness.④ Dec-remental responses were greater in patients with myasthenia gravis than that in ALS patients.Conclusion Decre-mental response with proximal muscle recording is not an uncommon feature in ALS patients;therefore,it should not be treated as a criterium to rule out ALS.Abnormal decremental response of trapezius muscle is found more fre-quent than that of deltoid muscle.Decremental response range in patients with myasthenia gravis is significantly lar-ger than that in ALS patients.One should be more careful when diagnosing ALS patients with fluctuating muscle weakness and abnormal decremental response.

Objective To compare the curative effect and complication of traditional open thyroid operation and minimally invasive laryngeal endoscopic thyroid operation.Methods 80 cases with thyroid disease were randomly divided into group I and groupⅡof 40 cases in each group,the group Ⅰ received the minimally invasive laryngeal endoscopic thyroid operation, while the Ⅱ group was treated with traditional open thyroid operation.The operation time,intraoperative amount of bleeding,postoperative drainage,hospitalization time and complications of two groups were compared.Results The operation time,hospitalization time of groupⅠwere (63.1 ±0.2)min,(5.7 ±0.6)d, which were significantly shorter than those of the control group[(90.4 ±1.1) min,(8.4 ±0.2) d].Amount of bleeding,postoperative drainage volume after operation in groupⅠwere (19.2 ±0.9) mL,(30.4 ±2.6) mL,which were significantly less than those of the control group[(42.1 ±1.8)mL,(46.2 ±1.2)mL] (t=9.819,12.168, 11.182,9.928,all P <0.05);The incidence of postoperative complications of group Ⅰ was 2.5%,lower than 15.0%of groupⅡ,the difference was statistically significant between the two groups(χ2 =13.197,P <0.05). Conclusion Minimally invasive laryngeal endoscopic thyroid operation has advantages in little trauma,short opera-tion time,rapid postoperative recovery.

Objective To compare the humoral immune response of BALB/c mice immunized by recombinant plasmids PeDNA3.1-M-NS1 and pcDNA3.1-N-NS1.Methods Dengue type 2 virus(DENV2)NS1 gene were constructed two partial sequences(1-413 bp)of the pcDNA3.1 eukaryotic plasmids and pET28a(+)plasmid for prokaryotic expression,identification,purification and quantification.The BALB/c mice were immunized by pcDNA3.1-M-NS1,pcDNA3.1-N-NS1 recombinant plasmids with adjuvant.Each animal received a primary inoculation and two boosts at 1-week intervals.Then the blood samples of BALB/c mice were collected from different experiment groups at day 7,14,28 and 56,respectively after first immunization.The specific IgM/IgG antibodies for NS1 protein in serum were confirmed by indirect ELISA.And then the activities of the specific protective antibody were determined by cytopathic effect inhibition(CPEI).Results Construction of the pET28a(+)-NS1 m/pET28a(+)-NS1n prokaryotic expression plasmid,SDS-PAGE analysis showed that,NS1 gene partial sequence was expressed,both the relative molecular weight of about 22.3×103:Western blot showed that the protein can bind anti-His tag monoclonal antibody;byNi affinity chromatographywith apurity of 92% protein,on the C6/36 cell toxicity,and can be used ELASA detection.The results showed that the levels of specific IgM/IgG antibody and neutralizing antibody activities were increased in pcDNA3.1-M-NS1 booster immunization group than other groups.The result had been observed longer duration of antibody level in peDNA3.1-M-NS1 booster immunization group.Conclusion Humoral immune response were significantly different between pcDNA3.1-M-NS1 and pcDNA3.1-N-NS1 recombinant plasmid immunized mice groups.

Objective To study the Mandarin long-term average speech spectrum (LTASS),the effects of gender,language and region material on LTASS were explored.This research would provide reference for clinical and academic use of Mandarin LTASS.Methods One hundred and sixty subjects from eight major dialectic regions of China,aging 16~30 (average 22.3±3.3),were asked to read two materials (storybook,newspaper)in Manda-rin at a steady speed and conversational level inside a standard soundproof booth.Ten males and ten females were in each dialectic region.A microphone (B&K 4192)was used to monitor each subject's speech which was analyzed with a real-time spectrum analyzer to obtain the long-term spectrum (B&K PULSE 3560C).Results Different Man-darin LTASS spectra were obtained based on gender,material and regions.The spectra were tested with profile a-nalysis.Language material had no significant effects on the Mandarin LTASS (F =2.52,P =0.11 ).There was, however,significant gender difference (F =116.39,P =0.00).Males had spoken with intensity levels higher than females at 100 Hz to 200 Hz.There was also a dialectic difference in the Mandarin LTASS (F =1.29,P =0.02). Speakers from Guangdong had the lowest intensity levels compared to those from other regions.Conclusion Genders and regions are factors that need to be considered when LTASS is used for clinical purpose and academic study.

Objective To explore the correlative factors of sudden death in patients suspected with pulmonary thromboembolism (PTE) in emergency room (ER).Methods A retrospective analysis was conducted.The clinical data of 12 patients with sudden death suspected with PTE (sudden death group) in ER of the Air Force General Hospital from January 2011 to June 2014 were analyzed.The non-sudden death group included 35 patients during the same time period who were diagnosed with PTE based on findings of CT pulmonary arteriography (CTPA) and showed no sudden death in ER.Factors,including sex,age,previous operation,tumor,syncope,dyspnea,bilateral or unilateral edema of lower extremity,heart rate (HR),white blood cell count (WBC),D-dimer,arterial partial pressure of oxygen (PaO2),arterial partial pressure of carbon dioxide (PaCO2) and typical clinical manifestation of electrocardiogram (SⅠTⅢQⅢ),were compared between the two groups.The potential predictors of sudden death of PTE were analyzed by logistic regression analysis.Results Young age (years old:51.3±15.5 vs.62.3±14.4),lower PaO2 [mmHg (1 mmHg =0.133 kPa):49.9± 12.3 vs.62.7± 10.2],higher HR (bpm:122.0± 19.5 vs.89.1 ± 18.5) and higher WBC (× 109/L:13.8 ± 6.9 vs.7.2 ± 2.5) were found in sudden death group as compared with those in non-sudden death group (P ＜ 0.05 or P ＜ 0.01).There was no significant differences in D-dimer level and PaCO2 between sudden death group and non-sudden death group [D-dimer (pg/L):986 (891,3 230) vs.2089 (598,3 397),PaCO2 (mmHg):33.0 (28.6,43.4)vs.36.5 (32.9,41.0),both P ＞ 0.05].The syncope,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months,bilateral asymmetrical edema in sudden death group were more than those of the non-sudden death group,and chest pain was less (P ＜ 0.05 or P ＜ 0.01).Difference in gender,dyspnea and typical SⅠTⅢQⅢ in electrocardiogram were not significant between the two groups (all P ＞ 0.05).It was shown by multiple logistic regression analysis that higher HR [odds ratio (OR) =1.124,95％ confidence interval (95％CI) =1.024-1.235,P =0.014] and higher WBC (OR =1.347,95％CI =1.043-1.738,P =0.022) were identified as independent risk factors of sudden death for PTE.Conclusions Gender,dyspnea,typical S Ⅰ TⅢQⅢ in electrocardiogram,PaCO2 and D-dimer seem unrelated to sudden death of patients with PTE.Young age,chest pain,syncope,bilateral asymmetrical edema,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months and low PaO2 were potential predictors of sudden death according to the univariate analysis.Higher WBC and higher HR are independent risk factors of sudden death for PTE patients.

Objective To determine the influence of depression on C reactive protein in patients with chronic ischemic heart failure (CIHF).Methods 73 patients with CIHF were divided into CIHF without depression group(n=49) and CIHF with depression group (n=24) according to the standard sore of the self-rating depression scale.C reactive protein(CRP)and other clinical characteristics were detected.The geometric means of CRP between two groups were compared,and stepwise multiple linear regression analysis was used to confirm the independent correlation factors of CRP.Results (1) The geometric means of CRP in CIHF without depression group and CIHF with depression group were (1.38± 1.29)mg/L and (1.78±1.53) mg/L respectively.No statistically significant difference was found between two groups(P=0.276).(2) Stepwise multiple linear regression analysis showed that aspirin inversely related with CRP(β=-1.070,P=0.004),but brain natriuretic peptide (BNP)positively related with CRP(β=0.426,P=0.001).Conclusion Depression has no additional effect on CRP in patients with CIHF.

Objective To analyze the efficacy of rosuvastatin on the patients with hyperlipidemia and hypertension.Methods From March 2011 to June 2012,112 cases with hyperlipidemia and hypertension in our hospital were enrolled in this study.Patients were randomly divided into treatment group and control group (56 patients,each).Patients in control group were treated with oral amlodipine 5 mg/d.Patients in treatment group were treated with oral rosuvastain 10 mg/d and oral amlodipine 5 mg/d.One month after the treatment,the levels of blood pressure,total cholesterol (TC),tryglyceride (TG),low density liporotein (LDL-C),high density lipoprotein cholesterol (HDL-C),high sensitivity C-reactive protein (hsCRP) were determined.The occurrence of adverse effects were observed.Results One month after treatment,systolic blood pressure and diastolic blood pressure were significantly decreased in both two groups compared with pre-treatment [Control group:(135.2±9.51)mm Hgvs.(59.2±7.3)mm Hg,(88.8±5.2)mm Hg vs.(99.5±8.3)mm Hg,t=4.95,2.87; Treatment group:(130.2±5.5)mm Hg vs.(160.3±9.3)mm Hg,(86.7± 10.2)mm Hg vs.(99.7±8.3)mm Hg,t=5.03,2.94,all P＜0.01],but more declines were found in treatment group than in control group(t=3.96,3.42,both P＜0.001).The levels of LDL-C,TG and TC were significantly decreased in both two groups compared with pre-treatment [Control group:(2.64±0.72)mmol/L vs.(3.97±0.84)mmol/L,(1.89±0.25)mmol/L vs.(2.56±0.45)mmol/L,(4.23±0.56)mmol/L vs.(7.36±0.48)mmol/L,t=2.58,3.03,2.36,P=0.013,0.004,0.022;Treatment group:(1.75 ± 0.68) mmol/L vs.(3.85 ± 0.79) mmol/L,(1.71 ± 0.18) mmol/L vs.(2.63±0.42)mmol/L,(3.18±0.47)mmol/L vs.(7.20±0.56)mmol/L,t=2.77,3.16,2.59,P=0.008,0.003,0.012,respectively],but more declines were observed in treatment group than in control group(t=6.73,4.37,10.70 respectively,all P＜0.05).The HDL-C concentrations were increased in both two groups compared with pre-treatment [Control group:(0.97±0.26)mmol/L vs.(0.75±0.31)mmol/L,t=2.89,P=0.006; Treatment group:(1.09±0.23)mmol/L vs.(0.72±0.24)mmol/L,t=3.01,P=0.004],but more increment were observed in treatment group than in control group(t=2.59,P＜0.05).The hsCRP concentration was significantly reduced in treatment group compared with pre-treatment [(1.32±0.17) mg/L vs.(4.97±0.13) mg/L,t=4.40,P＜0.001].There were no significant differences in liver and kidney function between the two groups.Serious adverse effects were not found.Conclusions Rosuvastatin combined with routine antihypertensive therapy can effectively decrease the levels of serum LDL-C,TG,hsCRP; increase serum HDL-C concentration and blood pressure can be effectively controlled.