Objective To observe the effect of acupuncture at neck Jiaji acupoints combined with moxibustion to cervical vertigo.MethodsIn Wuhan Chinese and Western Union Hospital from August 2015 to November 2016, 80 patients with cervical vertigo were randomly divided into two groups, the treatment group was treated by needling cervical Jiaji points, at the same time with moxibustion on neck Jiaji acupoints, the control group was given drug treatment.ResultsThe total effective rate in the treatment group was 95%, but the control group total effectiveness was 72.5%, the effect of the two groups had significant difference(P<0.05).ConclusionAcupuncture at Neck Jiaji points combined with moxibustion treatment had the functions of relieving rigidity of muscles and activating collaterals,dispelling cold and dredging channel blockage, relieveing local muscle spasm,improveing blood circulationand improve the function of the blood supply to the brain to vertigo.

Objective To explore the dynamic changes of endothelial barrier antigen (EBA) and vascular endothelial growth factor (VEGF) expressions in cerebral cortex under the condition of blood-brain barrier damage in rats following radi?ation-induced brain injury, which provided clinical references. Methods Forty-eight clean grade male SD rats were divid?ed into the control group and 7 d, 14 d, 28 d after brain irradiation group (n=12 for each group) by using stochastic indicator method. The radiation-induced brain injury model was established by using electronic computer X-ray tomography tech?nique. The 3%Evans blue (EB) was injected into rats according to the dose of 3 mL/kg via the tail vein, then the blood ves?sels of cerebral cortex were exposed after having a craniotomy. EB extravasation was detected by microcirculation micro?scope. The permeability of blood-brain barrier was evaluated by using microscope vascular camera device. The expressions of EBA and VEGF in the cerebral cortex were measured by immunohistochemistry staining in each group. Results Both of EB extravasation and VEGF expression in rat cerebral cortex were significantly increased in injury group at day 7, 14 and 28 after brain irradiation compared with those of control group (P<0.05), and which were gradually decreased from day 7 to day 28 after brain irradiation. There were significant differences in EB extravasation and VEGF expression between the injury subgroups (P<0.05). There was a positive correlation between EB extravasation and VEGF expression (r=0.898, P<0.001). The expression levels of EBA were decreased at different time points in injury groups compared with those of control group (P<0.05), and gradually increased from day 7 to 28 after injury. There were significant differences in expression levels of EBA between injury subgroups (P<0.05). The expression of EBA was negatively correlated with EB extravasation (r=-0.866, P<0.001). Conclusion The increases of blood-brain barrier permeability have important relation to the decreases of EBA expression and the increases of VEGF expression after radiation-induced brain injury.

Objective To investigate the morbidity,clinical feature,diagnosis and therapy of ovarian disease in children.Methods Eighty-two children with ovarian disease were admitted and treated in Nanjing children's hospital from Jan.1992 to Jan.2007,were analyzed retrospectively with age,emergency admissions or not,dwell,pathology and method of operation.Results The age of 82 patients ranged from 1 day to 14 years old and the mean age was 6.7 years old.Thirty-one cases(37.8%) were emergency admissions and 51 cases(62.2%) were routine admissions.Twenty-seven cases(32.9%) were rural patients and 55 cases(67.1%) were urban patients.Forty-five cases(54.8%) were nontumorous disorder,31 cases(37.8%) were benign tumor and only 6 cases(7.4%) were malignant tumor.About the morbidity,12 patients(14.6%)were admitted from 1992 to 1996,24 patients(29.5%) from 1997 to 2001 and 46 patients(55.9%) from 2002 to 2007.Chemotherapy were carried out in 6 cases with malignant tumor in internal medicine,2 cases with sexual precosity kept observation,the others were cured.Conclusions Ovarian disease can occur at any age in children.The clinical manifestation is characterized mainly by acute abdomen.The incidence of ovarian disease of children in urban areas is higher than that children in rural areas.The morbidity continues to show an upward tendency and the pathologic manifestations are mostly benign,laparoscopic operation has obviously superiority.

Objective To explore the feasibility of single port laparoscopy in classification and treatment of Meckel's diverticulum in children and its guiding treatment.Methods The clinical data in 75 children cases of Meckel's diverticulum with symptoms treated in our hospital from Aug.2011 to Aug.2015 were retrospectively analyzed.Meckel's diverticulum was classified under single port laparoscopy.The operation modes were selected according to different classifications.The excised materials were submitted to the pathologic examination.Results Among 75 children cases,50 cases were the simple type of Meckel's diverticulum and 25 cases were complex type of Meckel's diverticulum.The average operative time in the simple type and complex type was (38.93±8.75) min and(55.64 ± 13.27) min respectively,average bleeding amounts were (46.58 ± 15.81) mL and (50.12 [16.90) mL respectively,average postoperative hospitalization time was (7.33±1.41)d and (7.52 ± 1.68)d respectively,the operative time in the simple type was less than that in the complex type(P＜0.05),the other two indexes had no statistical difference between the two groups(P＞0.05).The ectopic gastric mucosal pathological change was only seen in the simple type,while the inflammatory manifestation in the complex type had higher proportion.The main clinical manifestations were lower gestational tract bleeding and infection.The two groups all obtained follow up.One case of simple type appeared the symptoms of abdominal pain and hematochezia and was cured after the second operation.Conclusion Meckel's diverticulum can be divided into the simple type and complex type under single port laparoscopy.The operation mode can be selected according to different types.This method is safe and reliable and is worthy of being clinically promoted.

Objective To study the effect of Ro60 on migration and invasion of lung adenocarcinoma A 549 cells. Methods We knocked down Ro60 and analyzed the ability of migration and invasion of A 549 cells.Quantitative real time ( RT)-PCR and Western blotting were performed to detect mRNA and protein levels of selected molecules associated with migration and invasion of neoplasm .Results When Ro60 was downregulated , the migration and invasion of A 549 cells decreased markedly.Meanwhile,the mRNA expression of matrix metalloproteinase (MMP)9,c-Src etc was observably reduced.Furthermore, downregulation of Ro60 diminished the expression of Src protein and the activation of MMP-9 protein.Conclusion Downregulation of Ro60 inhibits migration and invasion of A549 cells by regulating Src protein and activating MMP-9 protein.

AIM To determine the dynamic changes of monoamine neurotransmitters and their metabolites in various brain regions of cerebral ischemia reperfusion mice. METHODS Concentrations of monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT) and metabolites were determined by HPLC-ECD on d 0,1,3,5 and d 20 after cerebral ischemia reperfusion by common carotid artery occlusion. RESULTS The cerebral ischemia reperfusion mice showed decreased concentrations of NE, MHPG, DA, DOPAC, 5-HT and 5-HIAA in various brain regions, especially in hippocampus. CONCLUSION Several neuron systems play an important role in neurons damage of cerebral ischemia reperfusion, especially the NE and DA in hippocampus which is sensitive to the ischemia damage. The data offer useful guides for clinical treatments of cerebral ischemia diseases.

Objective To construct a eukaryotic expression plasmid vector encoding Cbl-b gene-specific short hairpin RNAs (shRNAs),and to evaluate its interference effect,so as to lay a foundation for further study on the role of Cbl-b in the immunotherapy of malignant melanoma.Methods According to the sequence of Cbl-b cDNA,4 pairs of shRNAs targeting the Cbl-b gene were designed and synthesized,and then inserted into the plasmid PGPU6/GFP/Neo to construct recombinant plasmids.After identification by DNA sequencing,the 4 shRNA expression vectors were cotransfected into 293T cells with the Cbl-b gene eukarytic expresson plasmid,respectively.The knockdown efficiency of these shRNA expression plasmids on Cbl-b expression was evaluated by real-time (RT) fluorescence-based quantitative PCR and Western blot at 48 hours aftert transfection.Results Sequencing analysis revealed that all the 4 pairs of shRNAs were successfully inserted into the eukarytic expression vector PGPU6/GFP/Neo.As RT-PCR and Western blot showed,all the 4 shRNA-expressing vectors could downregulate Cbl-b expession,and the NO.1 shRNA-expressing vector displayed the strongest interference effect(P ＜ 0.05).Conclusions A eukaryotic expression plasmid vector was successfully constructed for Cbl-b gene-specific shRNAs,and the most effective shRNA was selected in this study.

Objective To investigate the strategy and efficacy of enteral nutrition support of patients with spontaneous intraventricular hemorrhage-induced coma.Methods 139 patients were randomly divided into study group (treated with enteral nutrition mixed suspension,n =67) and control group (treated with normal full nutritional homogenized product,n =72) with a random number generating software.Enteral nutrition support was administered in 6-48 hours after admission.The total daily intake of enteral nutrition preparation was 1 000 ml (4 186.8 kJ),supplemented by liquid food.Body weight,serum albumin,serum total protein,hemoglobin,lymphocyte count,incidence of infection,level of consciousness and incidence of complications were compared between the two groups.Results In the third week after onset,the serum albumin [(32.1 ± 3.3) g/Lvs.(30.5±2.3) g/L,P=0.041],total protein [(62.2±3.2) g/Lvs.(56.9±2.7) g/L,P=0.039],and hemoglobin [(125.5 ±5.7) g/Lvs.(120.7 ±6.4) g/L,P=0.027] were significantly higher in the study group than in the control group.The Glasgow score in the second week in the study group was 13.1 ± 1.9,significantly higher than that in the control group (11.0 ±2.3) (P =0.037);the incidence of nosocomial infection was significantly lower in the study group than in the control group [17.9％ (12/67) vs.29.2％ (21/72),P =0.021];the proportion of patients with abnormal blood test results and that of patients having fever for more than 7 consecutive days were both significantly lower in the study group than in the control group [31.3％ (21/67) vs.38.8％ (28/72),P=0.042;37.3％ (25/67) vs.41.7％ (30/72),P =0.047].The two groups showed no significant difference in the incidence of intracranial infection after external ventricular drainage (P =0.235).Conclusion For patients with spontaneous intraventricular hemorrhage-induced brain dysfunction,enteral nutrition support with enteral nutrition suspension could effectively improve nutritional status,reduce complications,therefore conducive to recovery.

Objective To evaluate in vitro effects of specific small interfering RNA (siRNA)-silencing of the casitas B-lineage lymphoma b (Cbl-b)gene on immunocompetence of primary murine lymphocytes. Methods Spleens were resected from C57BL/6 mice, and splenic lymphocytes were sterily isolated and cultured in vitro. These lymphocytes were divided into 3 groups: silence group transfected with a Cbl-b-specific siRNA using the EntransterTM-R 4000 reagent, negative control group transfected with a negative control siRNA using the EntransterTM-R4000 reagent, blank control group receiving no treatment. After additional culture for 72 hours, ELISA was performed to measure levels of interferon γ(IFN-γ)and tumor necrosis factor α (TNF-α)in culture supernatants of lymphocytes. In addition, the Cbl-b gene-silenced lymphocytes were co-cultured with B16F10 melanoma cells to evaluate their immunocytotoxic effects on melanoma cells. Results Splenic lymphocytes were successfully isolated from C57BL/6 mice and cultured in vitro, and the Cbl-b-specific siRNA was also successfully transfected into the primary murine lymphocytes and effectively down-regulated the expression of Cbl-b gene in them. Compared with the negative control group and blank control group, the silence group showed significantly increased supernatant levels of IFN-γ and TNF-α(all P < 0.05). The immunocytotoxic effect of lymphocytes on melanoma cells was significantly stronger in the silence group than in the negative control group. Conclusion Cbl-b gene silencing can promote secretion of IFN-γ and TNF-α by murine lymphocytes, and enhance their immunocytotoxic effects on B16F10 melanoma cells in vitro.

Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06％ ± 6.37％ vs.100.00％ ± 10.42％ and 108.70％ ± 9.90％ at 24 hours,42.93％ ± 5.93％ vs.100.00％ ± 6.24％ and 168.00％ ± 2.98％at 48 hours,all P ＜ 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P ＜ 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P＜ 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P＜ 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P ＜ 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P ＜ 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30％ vs.0.421％ and 2.22％ at 24 hours,4.06％ vs.0.577％ and 0.691％ at 48 hours,all P＜ 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.