ObjectiveTo develop and apply a method for detecting the titer of varicella-zoster virus(VZV)neutralizing antibodies based on complement dependence,so as to improve the sensitivity of traditional plaque reduction neutralization assay for detection of the titer of VZV antibody.MethodsThe antigen(live attenuated varicella vaccine)and antibody(human VZV immunoglobulin)were mixed in different proportions and different incubation times. After neutralization,the antigen-antibody mixture was inoculated into human diploid cell 2BS strain cultured in a six-well plate. After 7 ～ 10 d of culture,the number of plaques was counted by Coomassie brilliant blue staining,and the 50% neutralizing antibody titer was calculated by Karber′s formula. Under the optimal neutralization conditions obtained,the effect of complement on the sensitivity of neutralization experiment was explored by changing the addition amount of complement(lyophilized guinea pig serum)to evaluate the optimal addition amount of complement. According to the determined neutralization test parameters,the neutralizing antibody titers of 12 anti-VZV mouse sera and 14 anti-VZV human sera were detected by using traditional plaque method and complement-dependent plaque method respectively.ResultsThe key parameters of the detection method were determined:the titer of VZV standard antigen was 500 ～ 1 000 PFU/mL;the proportion of complement added to the antigen-antibody neutralization system was 1∶10(v/v),and the neutralization condition was 37 ℃ for 1 h. Both the complement-dependent plaque method and the traditional plaque method were positive for anti-VZV mouse serum antibody,while the antibody titer detected by the traditional plaque method was generally lower,and the antibody level of mice inoculated with 2 doses of live attenuated varicella vaccine was significantly higher than that of mice inoculated with 1 dose(t = 0. 45,P < 0. 05);Both of the two methods were positive for anti-VZV human serum antibody.ConclusionA complement-dependent detection method for neutralizing antibody titer of VZV was established. The addition of complement significantly improved the sensitivity of neutralization detection. The evaluation of the titers of neutralizing antibodies in mouse serum with different immunization strategies by the method suggested that the immune effect of two doses of vaccine was better than that of one dose.

Objective To observe the clinical efficacy of acupuncture and moxibustion combined with cognitive training in treating cognitive impairment after traumatic brain injury (TBI). MethodsSixty patients were recruited into the study and randomly divided into the control group and the treatment group according to the MINIMIZE software. Patients in the control group were treated with cognitive training and regular treatment. Besides the traditional treatment, patients in the treatment group were additionally treated with acupuncture and moxibution. The treatment lasted four weeks. Mini-Mental State Examination (MMSE) and Loewenstein Occupational Therapy Cognitive Assessment (LOTCA) were applied to evaluate the patients’ cognitive function before and after the treatment.Results In the end, there were 27 patients in the control group and 28 patients in the treatment group, because 5 patients withdrew from the study. After treatment, scores of MMSE, LOTCA and their sub-items in the control group and the treatment group increased significantly (P<0.05), and the scores in the treatment group were higher than the control group (P<0.05).Conclusion Combination of acupuncture, moxibution, and cognitive training could help patients after TBI to increase the scores of MMSE and LOTCA, and improve the cognitive impairment caused by TBI. Its therapeutic effect is superior than the pure cognitive training.

OBJECTIVETo observe the clinical efficacy on cognitive impairment after traumatic brain injury (TBD treated with acupuncture and cognitive training.

METHODSSixty patients were randomized into an observation group and a control group, 30 cases in each one, and 5 cases of them were dropped out due to the earlier discharge. Finally, there were 28 cases in the observation group and 27 cases in the control group. In the control group, the cognitive training and conventional treatment were applied. In the observation group, on the basis of the treatment as the control group, acupuncture was applied to Baihui (GV 20), Fengchi (GB 20), Geshu (BL 17) and Fenglong (ST 40), once a day, for 4 weeks totally. The mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) were adopted to evaluate the cognitive function in the patients of post-TBI cognitive impairment.

RESULTS(1) After treatment, the total score in MMSE and the score of each item were increased significantly as compared with those before treatment in the two groups (all P<0. 05). Except for the score of immediate recall, the score in MMSE and the score of each of the other items were increased significantly in the observation group as compared with those in the control group after treatment (all P<0. 05). (2)After treatment, the total score in MoCA and the score of each item were increased significantly as compared with those before treatment in the two groups (all P<. 05). Except for the score of nomenclature item, the total score in MoCA and the score of each of the other items were increased significantly in the observation group as compared with those in the control group after treatment (all P<0. 05).

CONCLUSIONBoth the simple cognitive training and the combined therapy of acupuncture and cognitive training improve MMSE and MoCA scores and relieve the cognitive impairment induced by TBI. But the combined therapy achieves the much better efficacy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Brain Injuries ; psychology ; therapy ; Cognition ; Cognition Disorders ; psychology ; therapy ; Cognitive Therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

Objective To evaluate the effect of ultrasound-guided botulinum toxin A injection into the salivary glands in treating patients with sialorrhea attributable to brain damage.Methods Nine subjects with sialorrhea attributable to traumatic brain injury (n =5),cerebral infarction (n =3) and hypoxic ischemic encephalopathy (n =1) were recruited into the study.With the guidance of ultrasound,15 units (U) of botulinum toxin type A was injected into the highest and lowest points of the two parotid glands,and 20 U was injected into two points of both submandibular glands.The saliva flow rate and the severity and frequency of sialorrhea were recorded before the treatment and 1 week,4 weeks and 12 weeks later using the Drooling Scoring System (DSS).Results The saliva flow rate before treatment (0.49 ± 0.12 g/min) decreased significantly to 0.24 ± 0.08 g/min after 1 week,but then increased again to 0.28 ±0.12 g/min after 4 weeks and 0.34 ±0.11 g/min after 12 weeks.The average DSS severity score showed the same pattern:4.35 ± 0.48 initially 1.92 ± 0.37 after 1 week,1.92 ± 0.37 after 2 weeks and 2.32 ± 0.64 after 12 weeks.The DSS frequency score,however,remained significantly lower:3.56 ± 0.49 initially,2.01 ± 0.42 after 1 and 4 weeks and 2.28 ± 0.63 after 12 weeks.Conclusion Botulinum toxin type A can effectively improve the salivary secretion and sialorrhea attributable to brain damage.

Background Inflammation is one of the most popular aspects in the studies of diabetic retinopathy (DR) mechanisms.Researches showed that S100A8/A9 participate in the inflammatory procedure of many diseases,however,the relationship between S100A8/A9 complex and retinal inflammation of DR needs to be researched.Objective This study was to detect the serum S100A8/A9 level of diabetes mellitus (DM) and DR patients,and explore its role in DM an DR development.Methods A cases-controlled study was carried out.The DR patients,type 2 DM patients without retinal change and heathy controls were enrolled in Shanghai Xuhui Central Hospital from January to June 2014,and 30 patients for each group.The DR patients were subgrouped to non-proliferative DR (NPDR) group and proliferative DR (PDR) group.The periphery blood was collected to isolate the serum,and serum S100A8/A9 complex level was detected by ELISA.Serum high-sensitivity C-reactive protein (hsCRP) and glycosylated hemoglobin A1C (HbAlc) level was assayed by immunity turbidimetry and immune agglutination respectively.Results Serum S100A8/A9 complex levels in the DR group,DM group and normal control group were (9.74±0.59),(11.41 ±0.64) and (6.46 ±0.62) μg/L,respectively,and the serum S100A8/A9 complex level in the DM group and DR group was significantly higher than that in the normal control group,and the serum S100A8/A9 complex level in the DM group raised in compared with the DR group (all at P＜0.01).Serum hsCRP levels in the DR group,DM group and normal control group were (1.40±0.34),(1.27±0.13) and (1.11 ± 0.12)mg/L,respectively,with the highest value in the DR group and the lowest value in the normal control group (all at P=0.00).The serum HbAlc levels were higher in the DR group and DM group than those in the normal control group (both at P =0.00),while no significant difference was found in the serum HbAlc level between DR group and DM group (P =0.12).There was no significant differece in the serum S100A8/A9,hsCRP and HbAlc levels between NPDR group and PDR group (t=-0.10,P =0.92;t =-0.17,P =0.87;t =0.66,P =0.51).A weak positive correlation was seen between serum S100A8/A9 level and serum hsCRP level (r =0.36,P =0.00).Conclusions As an inflammatory marker,S100A8/A9 complex might play an important role in the pathogenesis and development of DR.Intensive control of glycemia can alleviate retinal inflammation in DM patients.

Objective To compare the effects of cobalt chloride (CoCl2) induced chemical hypoxia and hypoxia environment on the proliferation,migration and phenotype transformation of rat primary pulmonary artery fibroblasts (PAFs).Methods Primary PAFs were isolated and cultured.Cells were stimulated by CoCl2,or hypoxia cell culture (1% O2) was used to stimulate and induce PAFs.Then the effects of CoCI2 and hypoxia environment on PAFs were compared by CCK-8 assay,scratch assay,transwell assay,phenotype marker protein expression and PI3K/Akt signaling pathway protein expression.Results Compared with the control group,100 μmol/mL CoCl2 stimulation had no significant effect on the cell proliferation activity,cell migration ability and phenotype transformation ability of PAFs (P>0.05);while 1% O2 could significantly improve the cell proliferation and migration activities of PAFs as well as the upregulation of α-SMA expression (P<0.05).Conclusion There exist differences of effects between CoCl2 induced chemical hypoxia and hypoxia environment on promoting cell proliferation, cell migration and phenotype transformation in PAFs.

Objective To investigate the effect of rivanol induction on pregnancy termination for patients with placenta previa during midtrimester.Methods From January 2010 to December 2015,16 patients of placenta previa underwent pregnancy termination induced by rivanol during midtrimester were regarded as the observation group, and 22 patients with normal placental position were regarded as the control group.The delivery time,amount of postpartum hemorrhage within 24 hours,one-time success rate of induced abortion,caesarean due to massive haemorrhage and postoperative infection of the two groups were recorded to analyze the clinical effect of rivanol.Results There was no statistically significant differences in the success rate,delivery time and caesarean due to massive haemorrhage between the two groups(P>0.05).The amount of intrapartum and postpartum hemorrhage in the observation group was more than that of the control group,with a statistically significant difference(P<0.05),but it was less than 500 mL,which did not significantly increase the related risk for patients.Conclusion Induced abortion by rivanol is a simple,safe and effective method for patients with placenta previa during midtrimester with fewer side effects and less trauma,which is the preferred method for such patients.

Objective To observe the effect of electroacupuncture(EA) on the hippocampal expression of GluA1,pGluA1,CaMK Ⅱ and pCaMK Ⅱ in rats with vascular dementia(VD),so as to find out the underlying mo lecular mechanisms of EA in treating VD.Methods Thirty-two Wistar rats were randomly divided into a shamoperation group,a model group,a sham-acupuncture group,and an EA group (8 in each group).Permanent bilateral common carotid artery occlusion was performed to model vascular dementia in the model group,the shamacupuncture group and the EA group,while exposure but no occlusion of the bilateral common carotid were performed in the sham-operating group.Novel object recognition test was adopted to prove the establishment of VD rat model.All the rats were kept in an immobilization apparatus while receiving treatments.EA was applied ontoBaihui (GV20) and Zusanli (ST36) in EA group for 30 min,once daily for 7 days.Sham-acupuncture group were treated with needles inserted 0.5 mm superficially.And the sham-operation group and the model group were only immobilized.The protein expression of GluA1,pGluA1,CaMK Ⅱ and pCaMK Ⅱ in hippocampal tissue was detected by western blotting.Results The expression of GluA1 in the model group (1.216 ± 0.102) was significantly less than in the sham-operating group (1.918 ± 0.137) (P ＜ 0.05).The expression of GluA1 in the EA group (1.653 ± 0.169) was significantly higher than in the model group (1.216 ± 0.102) and in sham-acupuncture group (1.231 ±0.188) (P＜0.05).The expression of CaMKⅡ in the model group (1.516±0.392) was less than in the sham-operating group (2.187 ± 0.231) (P ＜ 0.05).The expression of CaMK Ⅱ in the EA group (2.733 ±0.387) was significantly higher than in the model group (1.516 ±0.392) and sham-acupuncture group (1.493 ±0.205) (P＜0.05).The expression ofpGluA1 in the model group (1.502 ±0.419) was less than in the sham-operating group (2.253 ± 0.244) (P ＜ 0.05).The expression of pGluA1 in the EA group (2.382 ± 0.308) was significantly higher than in the model group (1.502 ± 0.419) and the sham-acupuncture group (1.498 ± 0.223) (P ＜ 0.05).The expression of pCaMK Ⅱ in the model group (0.394 ± 0.227) was less than in the sham-operating group (0.667 ±0.175) (P＜0.05).The expression ofpCaMKⅡ in the EA group (1.189± 0.346) was significantly higher than in the model group (0.394 ± 0.227) and the sham-acupuncture group (0.408 ± 0.231) (P ＜ 0.05).Conclusion EA can enhance the protein expression and phosporylation of GluA1 and CaMK Ⅱ,causing silent synapses transforming into functional synapses,and consequently,long term potentiation was facilitated and cognitive impairment was improved by EA.

Objective To investigate the roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells.Methods Different phenotypes of dCK plasmids were transfected into MCF-7 cells by liposome transfection,including dCK-Vector,dCK-WT (wild type),dCK-S74A (non-phosphorylation) and dCK-S74E (hyper-phosphorylation).All these cells were irradiated by 0,2,4,6,8 Gy X-rays,respectively.The transcriptional and translational level of dCK were detected with real time-PCR and Western blot,respectively.Radiosensitivity was analyzed using cell counting kit (CCK-8) and colony formation assays.Monodansylcadaverine staining (MDC) and flow cytometry were used to detect autophagy and apoptosis,respectively.Results Four phenotypes of dCK cell models were established successfully.After irradiation,the cell viabilities of MCF-7 and dCK-Vector decreased significantly as compared with mock group (t =14.469 and 9.357,P ＜ 0.05),the cell viabilities of dCK-WT,dCK-S74A and dCK-S74E showed no changes (P ＞ 0.05).The total mortalities of dCK-WT and dCK-S74E decreased significantly as compared with dCK-Vector (x2 =3.857-3.971,P ＜ 0.05),but no changes in dCK-S74A cells (P ＞0.05).The apoptosis rates in dCK-S74A,dCK-Vector and control group were up-regulated after irradiation (t =-4.531,-3.688 and-7.076,P ＜ 0.05),and the irradiation-induced apoptosis was reversed in dCK-WT and dCK-S74E (66％ and 68％ of the increase level in dCK-Vector group).The autophagy in dCK-WT and dCK-S74E increased by 22％ and 26％ (t =-9.051 and-8.411,P ＜0.01),but no changes were observed in dCK-S74A,dCK-Vector and control groups (P ＞ 0.05).Conclusions The dCK-WT and dCK-S74E could reverse the irradiation-induced apoptosis,increase the autophagy occurence,and decrease the total mortality,indicating that the phosphorylation of dCK at Ser-74 sites is related to the radiosensitivity of MCF-7 cells.

Objective To assess right ventricular (RV) regional and global volume and systolic function in patients with right coronary artery disease by real time three-dimensional echocardiography(RT-3DE).Methods RT-3DE images were acquired in 20 normal controls (Group A),30 patients with right coronary artery stenosis(Group B) and 10 patients with RV myocardial infarction(Group C).Volumetric RT-3DE images were analyzed by use of TomTec software,which provided regional (inflow,body and outflow) and global end diastolic volume(EDV),end systolic volume(ESV),stroke volume(SV) and ejection fraction(EF).RT-3DE quantitative data were analyzed to test repeatability.Results Compared with group A and group B,EDV and ESV of body and global were larger in group C;ESV of inflow and outflow were larger (P ＜0.05).EDV of group C was larger than group B(P ＜0.05).Compared with group A and group B,the EF of inflow,outflow and global in group C were lower(P ＜0.05).The EF of body in group C were lower than group B(P ＜0.05).There was no significant difference in regional and global volume and EF between group A and group B(P ＞0.05).The intra-observer and inter-observer's variability was low and consistency was high.Conclusions In patients with RV myocardial infarction,RV regional and global volumes were enlarged and systolic function was impaired.Evaluating right ventricular regional and global volume and function in patients with right coronary artery disease by RT-3DE has high repeatability.