By using extracellular single unit recording technique, locally suppressive effects of a single dose of ketamine on sub-cutaneous (s. c. ) bee venom-induced increase in firing of wide dynamic-range (WDR) neurons in spinal dorsal horn were investi-gated on urcthane-chloralose anesthetized cats. Injection of bee venom s.c. into the cutaneous receptive field (RF) resulted in asingle phase of prolonged, persistently increased firing of WDR neurons over background activity for more than 1 h. Local pre-treatment with ketamine (100 mM, 0. 1 m l) into the center of RF where bee venom was injected produced a dramatic suppressionof the increased neuronal firing by 60% (3.10± 0.42 spikes/s, n= 5) when compared with saline pre-treated group (7.61 ± 0.17spikes/ s. n = 5 ). Moreover, local post-treatment with the same dose of ketamine also produced a profound suppression of the in-creased neuronal activity by 81% (1.51±0.06 spikes/s, n=5) when compared with the saline post-treated group (7.76±0.15spikes s, n=5). However, s.c. administration with the same dose of ketamine into a symmetrical region on the bee venom un-treated contralateral hindpaw produced no affection on the increased firing of the WDR neurons, suggesting that the suppressiveaction of local ketamine was not the result of systemic effects. The present result suggests that ketamine may exert its localantinociceptive effects mainly through the peripheral NMDA receptors in addition to its partially potential blocking effects onsodium and voltage-sensitive calcium channels.

Objective To investigate the impact of tanshinone Ⅱ A on the level of brain NMDAR1 protein in rats after cardiopulmonary resuscitation and its effects of brain function protection.Methods Seventy-eight SD male rats were randomly (random number) divided into three groups:group A (sham group,n =6),group B (control group,n =36) and group C (Tanshinone Ⅱ A intervention group,n =36).All animals were induced to be models of cardiac arrest by choking.The rats of group C received intravenous injection of Tanshinone Ⅱ A in dose of 15 mg/kg immediately at initiation of resuscitation,while rats of group B were intravenous injected same amount of normal saline instead.Brains tissues of all rats were taken at 1,6,12,24,48 and 72 h after the restoration of spontaneous circulation (ROSC).Immunohistochemical staining method was applied for measuring the levels of brain tissue NMDAR1 and Caspases-3,while water content of the brain was detected by wet and dry weight ratio.The experimentaldata were analyzed by using one-way ANOVA.Results (①)The level of brain NMDAR1 protein in group B increased at 1 h and reached its peak at 6 h after ROSC,then its level gradually declined and dropped below normal at 48 h,72 h,and there were significant difference in variation of NMDAR1 protein levels in comparison with the group A (P ＜ 0.05) ; the NMDAR1 protein levels at 1,6,12 h in group C were significantly lower than those in group B at the same intervals (P ＜ 0.01),but no significant differences were seen at 24,48,72 h (P ＞ 0.05).(②)The level of brain Caspases-3 in group B increased after ROSC,and reached its peak at 48 h after ROSC,then declined and maintained above normal at 72 h,and this variation was significantly different from that of the group A (P ＜ 0.01) ; while the levels of caspase-3 at 1,6,12,24 h in group C were significantly lower than those in group B (P ＜ 0.01),but thses differences at 48,72 h were still significant (P ＜ 0.05).(③)The water content of brain tissue in group B increased at 1 h and reached its peak at 24 h after ROSC,then gradually decreased from 48 h,but maintained above normal at 72 h,and this trend of variation was significantly different from that of group A (P ＜ 0.01 or P ＜ 0.05).Compared with group B,water content of brain tissue in group C decreased more significantly (P ＜ 0.01).Conclusions Tanshinone Ⅱ A down-regulates brain NMDAR1 protein level at early stage in rats as well as significantly inhibits the level of Caspases-3 thereby ameliorating brain edema after cardiopulmonary resuscitation.

Objective To explore the verification process for the analytic performance of the quantitative project of molecular di-agnosis.Methods Based onMedical laboratory accreditation criteria for quality and competence in the field of molecular diagnos-tics application note(CL-36)(2014)and the relevant documents published by Clinical and Laboratory Standards Institute (CLSI), the performance verification methodology of PCR detection for hepatitis b virus nucleic acid was achieved.for.Results The within-run precision of DNA detection for the hepatitis b virus was 0.109 and 0.105;and the between-run precisionwas 0.1 57 and 0.137. Compared with the reference laboratory,the regression equation was Y =0.947+0.343X ,and the linear correlation coefficient was 0.990.The linear range was 5.00-1.10 and thequantitative detection limit was 500 IU/mL.Hemolysis had no effect on the detec-tion of samples.Conclusion The laboratory with molecular diagnostic program should conduct analytic performance verification,and the appropriate method should be chosen to clear performance verification.Conclusion Clearing the performance indicators of de-tection projects has a very positive role in the clinical use of detection projects..

Objective To investigate protective effects of the hydrogen saturated saline on acute lung injury and pulmonary fibrosis in rats with paraquat (PQ) poisoning. Method Forty-eight Sprague-Dawley rats were randomly divided into 3 groups, namely control group, PQ poisoning group and intervention group (n = 16 rats in each group) . Animals in PQ poisoning group and intervention group were fed with PQ in dosage of 50 mg / kg. Rats of control group were fed equivalent amount of distilled water instead. One hour after administration of PQ, rats of intervention group were treated with 5 ml / kg hydrogen saturated saline injected intra-peritoneally twice a day until the rats were sacrificed. The rats of poisoning group and control group were treated with intra-peritoneal injection of equivalent amount of normal saline. Arterial partial pressure of oxygen ( PaO2 ), 8-hydroxy-2' -desoxyguanosine (8-OHDG) and transforming growth factor β1 (TGF-β1) of lung tissue were measured on the 3rd and 21st day after PQ administration. Quantitative data was expressed as mean ±standard deviation (x-±s) . SPSS version 12. 0 package was applied for variance analysis and SNK-q test, and statistical differences were considered significant when P ＜ 0. 05. Results ① PaO2 decreased significantly in poisoning group (9. 34 ± 0. 47 kPa) and intervention group (10. 30±0.62 kPa) compared with control group (11.87 +/- 0.42 kPa) on the 3d (P ＜0.01), and as intervention group was compared with poisoning group, there was a significant difference ( P ＜ 0. 05 ) . On the 21st day, PaO2 was still lower in poisoning group (8. 36 ±0. 51 kPa) and intervention group ( 10. 14 ±0. 27 kPa) than that in control group ( 11.87 +0. 24 kPa) (P ＜0. 01 and P ＜0. 05, respectively), and as intervention group was compared with poisoning group, there was a significant difference ( P ＜ 0. 01 ) . ②The levels of 8-OHDG in lung tissue increased significantly in poisoning group (23.58±7. 18 ng/ml ) and intervention group (9. 49± 2. 45 ng/ml) on the 3rd day after PQ administration compared with control group (7.71 + 1.96 ng/ml) (P＜0. 01 and P＜0. 05, respectively), and as intervention group was compared with poisoning group there was a significant difference ( p ＜0. 01 ) . There were no significant differences in 8-OHDG level found among the groups on the 21st day after PQ administration (P ＞ 0. 05 ) . ③ The level of TGF-β1 (measured by mean optic density, MOD) in lung tissue of rats in poisoning group ( 10. 11±2.49 MOD) and intervention group (8. 14 + 1.58 MOD) exhibited in higher levels than control group (5.93 + 1. 98 MOD) on the 3rd and (5.97 + 2. 35 MOD) on the 21st day after PQ administration (P ＜0. 01 and P ＜0. 05, respectively), and however, a lower level of TGF-β1 was observed in intervention group on 3d and 21d compared with poisoning group (P ＜0. 05 and P ＜0. 01, respectively) . Conclusions Hydrogen saturated saline can alleviate oxidative stress, mitigate oxidative damage and inhibit pulmonary fibrosis of lung induced by PQ intoxication.

Objective To observe and analyze the effects of pidotimod on the immunity of Treg/Th17 cells in the children with Henoch-Schonlein purpura (HSP).Methods 60 cases of HSP were selected and randomly divided into the pidotimod treatment group and the conventional treatment group,30 cases in each group.30 healthy children were selected as the control group.The con-ventional treatment group was given the routine therapy,while the pidotimod treatment group was added with oral pidotimod dis-persible tablets on the basis of the routine therapy.The ratios of Treg cell subsets and Th17 cell subsets,the plasma expression lev-els of interleukin-17 (IL-17),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interferon inducible protein-10 (IP-10) among 3 groups and the time for rash relief,remission rate of hematuria and proteinuria,recurrence rate within 1 month in HSP children were observed and compared.Results The ratios of Treg cell subsets and Th17 cell subsets after treatment had no statisti-cal difference between the pidotimod treatment group and the control group(P >0.05),while which had the statistical difference be-tween the conventional treatment group and the control group(P <0.05 );the plasma IL-17,IFN-γ,IP-10 and TNF-α expression levels had no statistical differences between the pidotimod treatment group and the control group(P >0.05),but the IL-17 and IFN-γexpression levels in the conventional treatment group were still significantly higher than those in the control group.the remission rate of simple hematuria in the pidotimod treatment group was significantly higher than that in the conventional treatment group, while the recurrence rate within 1 month after treatment was significantly lower than that in the conventional treatment group.Con-clusion The application of pidotimod therapy in the treatment of children with HSP can significantly correct the imbalance of Treg cell subsets and Th17 cell subset,decrease the expression levels of the cytokines such as IL-17 and IFN-γ,suppress the autoimmune reaction so as to achieve the effects of significantly alleviating the impairment of renal function and reducing the short term recur-rence rate,which is benefit for the improvement of prognosis in the children patients with HSP.

Objective To investigate the effects of different doses of gabapentin on streptozotocin (STZ)-induced diabetic neuropathic pain in rats.Methods Male SD rats aged 6 weeks weighing 180-200 g were used in this study. Diabetes ntellitus ( DM) was induced by intraperitoneal STZ 60 mg/kg and confirmed one week later by blood glucose =16.7 mmol/L before breakfast. The DM rats were randomly divided into 4 groups ( n = 6 each) : gabapentin groups received intraperitoneal gabapentin 30, 60 and 120 mg/kg twice a day (at 9:00 am and 3:00 pm) for 3 weeks respectively and control group received intraperitoneal normal saline 0.6 ml instead of gabapentin. The paw withdrawal threshold to von Frey filament stimulation was measured before and at 30, 60, 120, 180, 240 min after first gabapentin injection and once a week for 3 weeks. Results After gabapentin 60 and 120 mg/kg, the paw withdrawal threshold to mechanical stimuli was significantly increased and lasted for about 4 h. The analgesic effect peaked at 60 min after IP gabapentin injection. Normal saline and gabapentin 30 mg/kg had no significant analgesic effect. The degree of analgesia was significantly decreased at day 14 and 21 of treatment with gabapentin 60 and 120 mg/kg as compared with that at 60 min after gabapentin injection. Conclusion The hyperalgesia and allodynia in rats with diabetes mellitus can be effectively reversed by gabapentin 60 and 120 mg/kg,while long-term use of gabapentin can induce drug tolerance.

Objective To investigate the effect of heroin abuse on attention switching. Methods Thirty-six Heroin abusers (33 males, 3 females) and 36 controls (32 males, 4 females) were enrolled in the study. Their cognitive function was tested by using the Switching Task, including Sustained Attention trials and Switching Attention trials. The reaction time and accuracy were recorded separately by the computer. Results The accuracy or reaction times were not signifi-cantly different between Switching Attention trial and Sustained Attention trial in heroin abusers, suggesting a lower Switch Costs value compared to the healthy controls [(19.7 ± 66.8) ms vs. (85.1 ± 92.4) ms]. The healthy controls showed faster reaction speed [Sustained Attention trial (695.3 ± 95.9) ms vs. Switching Attention trial (780.3 ± 93.3) ms, P<0.05] and higher accuracy [Sustained Attention trial (98.0%±2.2%) vs. Switching Attention trial (93.8%±5.0%), P<0.05] under the Sustained Attention trial. Compared with the healthy controls, the heroin abusers showed slower reaction speed [(791.6 ± 74.3) ms vs. (695.3±95.9) ms, P<0.05] and lower accuracy [(92.5%±8.4%) vs. (98.0%±2.2%), P<0.05] in Sus-tained Attention trial, but not in Switching Attention trial. Conclusions The present study has revealed absence of Switch Costs in heroin abusers, which may be related to the damage of heroin abusers in their Sustained Attention function.

Resveratrol is a natural phytoalexin with special pharmacological and health functions. Stilbene synthase (STS) is a key and rate-limiting enzyme in the biosynthesis of resveratrol that is present only in a limited number of plants. The content of resveratrol from Polygonum cuspidatum is more than 1000 times higher than grapes and peanuts. We speculate that the catalytic ability of different STS may be one of the reasons causing differences in the content of resveratrol. To verify the above speculation, Vitis vinifera stilbene synthase gene (VvSTS) was amplified according to overlap PCR protocol with genomic DNA as template. VvSTS and PcSTS (PcPKS5) were analyzed through heterologous expression in Escherichia coli. The expression products were purified with Ni-NTA sepharose affinity chromatography and desalted through PD-10 column. The molecular weight of the two fusion proteins was about 43 kDa. Enzyme reaction and product analysis showed that the two products were resveratrol. The enzyme kinetic analysis showed that the catalyze efficiency (Kcat/Km) of PcPKS5 was 2.4 times of the VvSTS. Our findings confirms that STS from certain plants has much higher catalytic capability.
Acyltransferases ; metabolism ; Fallopia japonica ; enzymology ; Recombinant Fusion Proteins ; biosynthesis ; Stilbenes ; metabolism ; Vitis ; enzymology

Objective It has been confirmed that Epstein-Barr virus ( EBV) is associated with the occurrence and development of the nasopharyngeal carcinoma ( NPC ) . We investigated the clinical significance of plasma concentrations of EBV-DNA in patients with NPC. Methods Since October,2013 to December,2016,471 patients were analyzed. The significantly associated between EBV-DNA before treatment and staging, tumor burden was analyzed. The survival rate of EBV-DNA before and after treatment was calculated. Results The median copies of pretreatment plasma EBV-DNA in patients is 137 copies,( range 0-494000) ,which is correlated with T stage,N stage,M stage,clinical stage and tumor burden load and that is statistically significant. Overall survival ( OS,P=0. 007) ,progression-free survival ( PFS,P=0. 011) and distant metastasis-free survival ( DMFS,P=0. 003) were significantly lower among patients with pretreatment plasma EBV-DNA more than 1300 copies/ml. Patients with detectable plasma EBV-DNA had significantly worse OS (P=0. 016),PFS (P=0. 000) and DMFS (P=0. 000) than patients with undetectable EBV-DNA after treatment. Cox multivariate analyze suggests that T stage and EBV-DNA after treatment were independent prognostic factors for OS,however the plasma EBV-DNA after treatment ( P=0. 006,0. 001) and N stage ( P=0. 037,0. 017) were independent prognostic factors for PFS and DMFS. Conclusions The plasma EBV-DNA level was significantly correlated with staging and tumor load before treatment in patients with NPC,and the prognosis of patients with higher copies before treatment could be worse. The plasma EBV-DNA after treatment is predictive for OS,PFS and DMFS.

Objective: To analyze the clinical features, diagnosis and prognosis of patients with primary lymphoepithelial carcinoma of the parotid gland. Methods: Clinical data of 13 patients diagnosed with lymphoepithelial carcinoma of the parotid gland in our hospital from 2009 to 2017 were retrospectively analyzed. The median follow-up time was 38.5 months. All patients received radiotherapy after operation. Results: Of 13 patients, 9 cases were male and 4 female. The median age was 33 years. At the initial diagnosis, 9 cases had primary lesions limited to the parotid gland, and 4 cases of lymph node metastases located in Ⅰ_b and Ⅱ regions of the neck. According to UICC2010 staging, 1 case was classified as stage Ⅰ, 1 as stage Ⅱ, 6 as stage Ⅲ and 5 as stage Ⅳ, respectively. Eleven surgically pathological specimens were tested with EBER in-situ, and 10 cases were positive for EBER. No patient died in the whole group. The 3-year overall survival rate was 100%. The 3-year progression-free survival rate was 76%. The 3-year local control rate was 92%. The 3-year metastasis-free survival rate was 84%. Conclusions The incidence of lymphoepithelial carcinoma of the parotid gland is relatively low. The pathological features are associated with EB virus. It is prone to present with cervical lymph node metastasis. The possibility of lymph node metastasis of nasopharyngeal carcinoma to the parotid gland should be excluded before treatment. At present, surgery combined with postoperative radiotherapy is the main treatment. The overall survival is favorable. Local recurrence and distant metastasis are the main causes of treatment failure.