Effects of nucleus accumbens GABA-lateral hypothalamic area MCH neural pathway on rewarding feeding / 中华行为医学与脑科学杂志

Objective:To explore the effects of the γ-aminobutyric acid(GABA) neurons and melanin-concentrating hormone (MCH) neurons of the nucleus accumbens (NAc)-lateral hypothalamic area (LHA) neural pathway on the rewarding feeding(palatable food sweat condensed milk) in the obesity rats.Methods:Total 142 male Wistar rats of SPF grade were divided into normal diet (ND) group ( n=68) and high-fat diet induced obesity (DIO) group ( n=74) according to the principle of body mass matching. The rats in the two groups were given normal diet and high-fat diet for 8 weeks. Eight weeks later, 6 DIO rats were randomly selected to observe the nerve projection from GABA neurons in NAc to MCH neurons in LHA by fluorogold retrograde tracing combined fluorescence immunohistochemistry. And the expressions of c-Fos and MCH in LHA after ingestion of sweet condensed milk(rewarding feeding) were observed by fluorescence immunohistochemistry (6 rats in each group). GABA receptor agonist Musimol or GABA receptor antagonist Bicuculine was microinjected into the nucleus of LHA to observe the effect of GABA on rewarding food intake in ND and DIO rats ( n=8 in each group), and the changes of rewarding food intake after blocking MCH signal ( n=8 in each group). SPSS 17.0 was used for statistical analysis, two-way ANOVA and post hoc Bonferroni test were used for comparison among multiple groups, and t-test was used for comparison between two groups. Results:After 8 weeks of high-fat diet modeling, the intake of delicious food in DIO rats was significantly higher than that in ND rats((12.52±2.29) mL, (7.45±1.23) mL, t=4.778, P<0.01) after satiety.The results of fluorogold retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neurons in NAc projected nerve fibers to neurons in LHA, and GABA A receptors in some neurons in LHA coexisted with MCH.The results of NAc-LHA pathway on delicious food intake showed that the interaction between rat group and drug intervention was significant( F=9.869, P<0.01). Simple effect analysis showed that the intake of delicious food after microinjection of Musimol into LHA nucleus of ND rats was significantly lower than that of microinjection normal saline ((4.25±1.38) mL, (7.29±1.49) mL, P<0.01), while the intake of delicious food after injection of Bicuculine was significantly higher than that of microinjection normal saline((10.72±2.11) mL, (7.29±1.49) mL, P<0.05). The intake of delicious food after microinjection of Musimol into LHA nucleus in DIO group was significantly lower than that of microinjection normal saline((3.51±1.77)mL, (13.68±2.95) mL, P<0.01), but there was no significant difference between microinjection Bicuculine and microinjection normal saline ((14.83±3.44) mL, (13.68±2.95) mL, P>0.05). The results of blocking MCH signal on delicious food intake showed that the interaction effect between SNAP-94847 and Bicuculine intervention was not significant ( F=1.468, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=15.880, P<0.01)and the main effect of Bicuculine intervention was significant ( F=6.930, P<0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of ND rats was significantly less than that of injection normal saline((4.78±1.72) mL, (7.63±2.77) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((6.24±2.18) mL, (4.78±1.72) mL, P>0.05). In the DIO rats, the interaction effect between SNAP-94847 and Bicuculine intervention was not significant( F=0.006, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=18.46, P<0.01) and the main effect of Bicuculine intervention was not significant ( F=2.059, P>0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of DIO rats was significantly lower than that of injection normal saline((6.89±2.11) mL, (12.19±4.36) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((8.72±2.26) mL, (6.89±2.11) mL, P>0.05). Conclusion:GABAergic signal in NAc can regulate the expression of MCH in neurons of LHA. In the DIO rats, the sensitivity of MCH neurons in LHA to satiety signal decreases and the hedonic feeding increases.

Regulatory effect of edaravone on the photoreceptor autophagy at the early stage of experimental retinal detachment in rats / 中华实验眼科杂志

Objective:To investigate the effect of edaravone, a free radical scavenger, on the regulation of retinal autophagy and the protection of photoreceptor cells at the early stage of experimental retinal detachment (RD) in rats.Methods:Fifty-one adult male Sprague-Dawley rats were used for RD model establishment, and another 24 rats were served as PBS injection group.The RD model was established via subretinal injection of 0.5% sodium hyaluronate into the right eye of the rats and the rats with successful modeling were randomly divided into RD model group and edaravone treatment group.The rats in the edaravone treatment group were given edaravone of 3 mg/kg intraperitoneally, twice a day after modeling, and the rats in the PBS injection group and RD model group were given equal volume of normal saline.Rats were sacrificed on the 1st day, 3rd day and 7th day following modeling.The T-superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in the intraocular fluid was detected.The expression levels of superoxide dismutase 2 (SOD2), nuclear factor E2-related factor 2 (Nrf2), autophagy related gene 4 (Atg4), microtubule-associated protein 1 light chain 3B (LC3B) and other proteins in retinal tissue were identified by Western blot analysis.TUNEL staining was performed on paraffin sections of the whole eyeball to analyze the apoptosis of photoreceptor cells.The study protocol was approved by an Ethics Committee of Xi'an Fourth Hospital (No. 2016016). The use and care of animals complied with the Regulations on the Administration of Experimental Animals.Results:The RD area was more than 60% in rat eyes of RD model.There were significant differences in MDA content and T-SOD activity among different groups at various time points (MDA: Fgroup=385.513, P<0.01; Ftime=13.021, P<0.01.T-SOD: Fgroup=48.865, P<0.01; Ftime=7.700, P=0.003). Compared with the PBS injection group, the MDA concentration was significantly increased and the T-SOD activity was significantly decreased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05). The MDA concentration was significantly reduced and the T-SOD activity was significantly elevated in the edaravone treatment group on the 1st, 3rd and 7th day after modeling in comparison with those of the RD group (all at P<0.05). Compared with the PBS injection group, the relative expression levels of SOD2 and Nrf2 proteins were significantly increased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased on the 1st, 3rd and 7th day after modeling (all at P<0.05). The expression level of SOD2 in the edaravone treatment group was significantly higher than that in the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and the expression level of Nrf2 was significantly increased in the edaravone treatment group on the 1st and 3rd day after modeling compared with that of the RD group (both at P<0.05), and the expression levels of Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased in the edaravone treatment group on the 3rd day after modeling in comparison with those of the RD group (both at P<0.05). No significant TUNEL positive cells were observed in PBS injection group at all time points, and TUNEL positive cells were observed on the 1st, 3rd and 7th day after modeling in the RD group, and the expression level of caspase-3 in the RD group was significantly increased in comparison with that of the PBS injection group ( P<0.05). The apoptosis of photoreceptor cells and the expression level of caspase-3 in edaravone treatment group were significantly decreased in comparison with those of the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05). Conclusions:The intraperitoneal injection of edaravone, twice a day, can significantly improve the antioxidant capacity of the retina after experimental RD in rats, regulate retinal autophagy and reduce the apoptosis of photoreceptor cells in early-stage RD.

Effects of LHA-NAc orexinergic pathway on gastric motility and reward feeding in rats / 中华行为医学与脑科学杂志

Objective:To investigate the regulation of orexinergic pathway from the lateral hypothalamic area (LHA) to the nucleus accumbens (NAc) on gastric function and reward feeding.Methods:(1)Forty-eight rats were randomly selected and divided into six groups: normal saline (NS) group, 1 μg orexin-A group, 5 μg orexin-A group, 10 μg orexin-A group, 20 μg orexin receptor antagonist (SB334867) group, 20 μg SB334867 + 5 μg orexin-A group with eight in each group according to the random number table. The gastric motility of rats was observed by injecting orexin-A and SB334867 into NAc. (2)Thirty-two rats were randomly selected and divided into four groups according to the random number table, with eight in each group. They were divided into NS + sham stimulation (SS) group, NS + electrical stimulation (ES) group, SB334867(20 μg) + SS group, and SB334867(20 μg) + ES group. The gastric motility of rats were observed by electro-stimulation of rat LHA and rat NAc injection of SB334867. (3)In order to observe the feeding-behavior related conditioned place preference (CPP) and gastric function (such as gastric emptying and gastric secretion), thirty-two rats were randomly divided into four groups with eight in each group by using the method of NAc injection of orexin-A and SB334867 according to the random number table: NS group, orexin-A(5 μg), SB334867(20 μg), SB334867(20 μg) + orexin-A(5 μg). (4)Thirty-two rats were randomly selected in accordance with the random number table and divided into four groups with eight in each group: NS + SS group, NS + ES group, SB334867(20 μg) + SS group, SB334867(20 μg) + ES group, using electro-stimulation of rat LHA and rat NAc injection of SB334867, observing the feeding-behavior related CPP and gastric function (such as gastric emptying and gastric secretion).Results:(1)In the gastric motility experiment, both the NAc microinjection of orexin-A and electrical stimulation of the LHA significantly increased the amplitudes and frequencies of gastric contraction in rats, and these effects could be blocked by the pre-administration of SB334867 (10 min after administration of orexin-A: 10 μg orexin-A group (60.78±5.67)% vs NS group (7.35±1.08)%; t=26.18, P<0.05). (2)The results of gastric emptying showed that the rates of gastric emptying were significantly increased by the NAc microinjection of orexin-A and electrical stimulation of LHA, which were blocked by the SB334867 pretreatment (after electrical stimulation of LHA: NS + SS group (71.18±17.78)% vs NS+ ES group (132.23±31.18)%; t=4.81, P<0.05). (3)Orexin-A microinjection in the NAc and electrical stimulation of the LHA significantly increased gastric acid secretion, and these effects could be blocked by pre-injection of SB334867 in NAc (90 minutes after administration of orexin-A: orexin-A group(100.18±23.23) vs NS group (39.23±7.69); t=7.05, P< 0.05) in the gastric secretion experiment.(4)The results of CPP showed that the rats were kept in the chocolate compartment for a longer time after the NAc microinjection of orexin-A and electrical stimulation of the LHA, which could be blocked by the SB334867 pretreatment in NAc (after LHA was electrically stimulated: NS+ SS group (36.23±6.23)% vs NS+ ES group (53.36±6.66)%; t=5.31, P<0.05). Conclusion:There is an orexinergic pathway from LHA to NAc that may regulate gastric function and food reward.

Clinical study of ultrasound evaluation of fiberoptic bronchoscope guided tracheotomy / 中华危重病急救医学

Objective:To explore the clinical effect of ultrasound evaluation of fiberoptic bronchoscope (FB) guided tracheotomy, which can provide help for difficult tracheotomy and new operators.Methods:The operating protocol was standardized for ultrasound evaluation of FB guided tracheotomy. Ten patients with difficult tracheotomy admitted to the department of critical care medicine of Donge Hospital from October 2019 to January 2020 were enrolled. According to this protocol, FB guided tracheotomy was performed under the ultrasound evaluation, and the amount of blood loss, operation time and related complications during procedures were collected.Results:The preparation of supplies, personnel, patients and the operation, the process of FB guided tracheotomy under the ultrasound evaluation were standardized. When tracheotomy was preformed for patients with difficult tracheotomy, it was necessary to use ultrasound first to evaluate the neck condition and vascular disorientation of patients, and the tracheotomy plan (tracheotomy site, incision size, and incision depth) was designed, and then the tracheotomy process was monitored under the guidance of FB. Among the 10 patients with difficult tracheotomy, 6 were male and 4 were female; body mass index was (32.2±1.4) kg/m 2. Tracheotomy was successfully completed within 10 minutes in all the 10 patients, with less than 5 mL blood loss, and no complications occurred. Conclusion:Ultrasound evaluation of FB guided tracheotomy can improve the clinical operations and ensure patient safety.

Effects of Wenxin granules and bisoprolol fumarate combined with aerobic exercise on serum matrix metallo-proteinase -1 and N - terminal pro - brain natriuretic peptide levels in patients with unstable angina pectoris / 中国基层医药

Objective To investigate the influence of Wenxin granules and bisoprolol fumarate combined with aerobic exercise on serum matrix metalloproteinase -1(MMP -1) and N - terminal pro - brain natriuretic peptide (NT - proBNP) levels in patients with unstable angina pectoris. Methods From January 2016 to February 2018, 92 patients with unstable angina pectoris in Lishui Hospital of Traditional Chinese Medicine were selected and randomly divided into control group(n = 46) and study group(n = 46) according to the digital table. On the basis of routine intervention,the control group was given bisoprolol fumarate for 2 weeks and aerobic exercise for 1 month, while the study group was given bisoprolol fumarate,aerobic exercise and Wenxin granules for 2 weeks. The frequency and duration of angina pectoris before and after 1 month of treatment,clinical efficacy,serum MMP -1 and NT - proBNP levels,blood rheology index(plasma viscosity,whole blood high shear viscosity,total blood low - cut viscosity,fibrinogen) and incidence of adverse reactions were observed. Results After 1 month of treatment,the frequency of angina pectoris was lower than that before treatment,and the duration was shorter than that before treatment,which in the study group [(2. 09 ± 0. 65)times/ week,(2. 24 ± 0. 83)min/ time] were better than those in the control group(t1 = 10. 898,t2 =9. 302,all P < 0. 05). The total effective rate of the study group(91. 30％ ) was higher than that of the control group (73. 91％ )(χ2 = 4. 842,P < 0. 05). After 1 month of treatment,the serum levels of MMP - 1 and NT - proBNP in the two groups were lower than those before treatment,which in the study group[(196. 10 ± 55. 01) g/ L,(403. 82 ± 114. 15)ng/ L] were lower than those in the control group ( t1 = 3. 619, t2 = 4. 185, P < 0. 05). At the end of treatment,the plasma viscosity,whole blood high - viscosity,whole blood low - cut viscosity,fibrinogen level in the two groups were lower than those before treatment,which in the study group were lower than those in the control group(all P < 0. 05). The incidence rate of adverse reactions in the study group was 15. 22％ ,which in the control group was 10. 87％ ,there was no statistically significant difference between the two groups (χ2 = 0. 383,P > 0. 05). Conclusion The combination of Wenxin granules, bisoprolol fumarate and aerobic exercise for patients with unstable angina pectoris not only can reduce the frequency of angina pectoris,MMP - 1 and NT - proBNP levels,but also can improve the treatment of disease. Besides,it does not increase the risk of adverse reactions.

Study on the polymorphism of PPAR-γgene and preeclampsia susceptibility / 中国医师杂志

Objective To investigate the correlation between peroxisome proliferator-activated re-ceptor-gamma ( PPAR-γ) gene polymorphism and preeclampsia ( PE) . Methods 110 PE patients and 110 normal pregnants who delivered in our hospital from May 2016 to October 2018 were selected as the study subjects. Detection of two loci of PPAR-γ gene by SNaPshot technique: rs10865710 and rs4684847. Re-sults ⑴ rs10865710:The genotype frequencies of CC, CG and GG in the control group were 44. 55％, 42. 73％ and 12. 73％, respectively, and those in the PE group were 31. 82％, 44. 55％ and 23. 64％, re-spectively. There was a critical difference in the distribution of genotype frequencies between the two groups (x2 =5. 975, P=0. 050); The frequencies of C and G alleles were 65. 91％ and 34. 09％ in the control group, 54. 09％ and 45. 91％ in the PE group, respectively. There was significant difference in the fre-quency distribution of C and G alleles between the two groups (x2 =6. 402, P=0. 015). ⑵ rs4684847:the genotype frequencies of CC, CT and TT in control group were 94. 55％, 5. 45％, 0, and those in PE group were 84. 55％, 15. 45％ and 0, respectively,with significant difference in the distribution of genotype frequencies between the two groups (x2 =5. 875, P =0. 015). The frequencies of C and T alleles in control group were 97. 27%, 2. 73% and those in PE group were 92. 27% and 7. 73%, respectively,with significant difference in allele frequency distribution between the two groups (x2 = 5. 551, P = 0. 030). ⑶ The frequency of GG genotype and G allele at rs10865710 locus in PE group was significantly higher than that in control group [OR (95% CI) =2. 600 (1. 190 - 5. 679), P = 0. 021; OR (95% CI) = 1. 64 (1. 117 -2. 411), P =0. 015]. The frequencies of CT genotype and T allele at rs4684847 locus in PE group were significantly higher than those in control group [OR (95% CI) =3. 168 (1. 199 -8. 374), P =0. 026; OR (95% CI) =2. 987 (1. 155 -7. 726), P =0. 030]. Conclusions The single nucleotide polymorphisms of PPAR-γ gene rs10865710 and rs4684847 may be related to the susceptibility to preeclampsia in Chinese population.

Study on the mechanism of microRNA-210 regulating the immune pathogenesis of Treg cells in pre-eclampsia / 中国医师杂志

Objective The aim of this study was to evaluate the number of Treg cells in preeclamp-sia (PE) patients, to explore the expression levels of microRNA-210 (microRNA-210) and forkhead box p3 (Foxp3) genes in preeclampsia, and to reveal the regulatory mechanism of microRNA-210 and Foxp3 in preeclampsia. Methods Serum levels of cytokines [ interleukin ( IL)-6, IL-10, IL-17, and transforming growth factor-beta 1 (TGF-β1)] were detected with enzyme-linked immunosorbent assay (ELISA). 29 pa-tients with late-onset preeclampsia (≥36 weeks of gestation) , 27 pregnant women with normal uncomplicat-ed pregnancies (≥36 weeks of gestation) and 20 healthy non-pregnant women were enrolled in the study. Reverse-transcription polymerase chain reaction ( qRT-PCR) was performed to detect mRNA expression for maternal placenta retinoic acid-related orphan receptor C (RORc), Foxp3, and miR-210. Foxp3 protein expression was evaluated by Western blot. Results ⑴The serum levels of IL-6, IL-17 and TGF-beta 1 in preeclampsia patients were significantly higher than those in normal pregnant women, and the level of Treg cytokine IL-10 was lower than that in normal pregnant women ( P <0. 05 ) . ⑵ The percentage of CD4 +CD25 +CD127 - /CD4 +T cells in peripheral blood of preeclampsia patients was significantly lower than that of normal pregnancy group and healthy non-pregnant women ( P <0. 001 ) . ⑶ The mRNA expression of Foxp3 in placenta of preeclampsia patients was significantly lower than that of normal pregnant women, RORc in preeclampsia patients was significantly higher than that of normal pregnant women, and the expres-sion of microRNA210 in preeclampsia patients was enhanced ( P<0. 01 ) . ⑷ Consistent with mRNA ex-pression results, lower protein expression levels of Foxp3 was observed in patients with PE compared with normal pregnant subjects. Conclusions Treg cells decreased in preeclampsia patients and Treg/Th17 im-balance existed in preeclampsia patients, which regulate maternal immune tolerance to fetuses. The expres-sion of Foxp3 in placenta of preeclampsia patients was significantly decreased, which was correlated with the expression of microRNA-210.

Effects of Wenxin granules and bisoprolol fumarate combined with aerobic exercise on serum matrix metallo-proteinase-1 and N-terminal pro-brain natriuretic peptide levels in patients with unstable angina pectoris / 中国基层医药

Objective To investigate the influence of Wenxin granules and bisoprolol fumarate combined with aerobic exercise on serum matrix metalloproteinase－1(MMP－1) and N－terminal pro－brain natriuretic peptide (NT－proBNP) levels in patients with unstable angina pectoris.Methods From January 2016 to February 2018, 92 patients with unstable angina pectoris in Lishui Hospital of Traditional Chinese Medicine were selected and randomly divided into control group(n=46) and study group(n=46) according to the digital table.On the basis of routine intervention,the control group was given bisoprolol fumarate for 2 weeks and aerobic exercise for 1 month, while the study group was given bisoprolol fumarate,aerobic exercise and Wenxin granules for 2 weeks.The frequency and duration of angina pectoris before and after 1 month of treatment,clinical efficacy,serum MMP－1 and NT－proBNP levels,blood rheology index(plasma viscosity,whole blood high shear viscosity,total blood low－cut viscosity,fibrinogen) and incidence of adverse reactions were observed.Results After 1 month of treatment,the frequency of angina pectoris was lower than that before treatment,and the duration was shorter than that before treatment,which in the study group [(2.09 ± 0.65)times/week,(2.24 ± 0.83)min/time] were better than those in the control group(t1=10.898,t2= 9.302,all P<0.05).The total effective rate of the study group(91.30% ) was higher than that of the control group (73.91% )(χ2 =4.842,P<0.05).After 1 month of treatment,the serum levels of MMP－1 and NT－proBNP in the two groups were lower than those before treatment,which in the study group [(196.10 ± 55.01) g/L,(403.82 ± 114.15)ng/L] were lower than those in the control group ( t1 =3. 619, t2 =4. 185, P <0. 05 ). At the end of treatment,the plasma viscosity,whole blood high－viscosity,whole blood low－cut viscosity,fibrinogen level in the two groups were lower than those before treatment,which in the study group were lower than those in the control group(all P<0.05).The incidence rate of adverse reactions in the study group was 15.22% ,which in the control group was 10.87% ,there was no statistically significant difference between the two groups (χ2 =0.383,P>0.05).Conclusion The combination of Wenxin granules, bisoprolol fumarate and aerobic exercise for patients with unstable angina pectoris not only can reduce the frequency of angina pectoris,MMP－1 and NT－proBNP levels,but also can improve the treatment of disease.Besides,it does not increase the risk of adverse reactions.

Decreased Expression of TRPV4 Channels in HEI-OC1 Cells Induced by High Glucose Is Associated with Hearing Impairment

PURPOSE: Previous reports have shown that hyperglycemia-induced inhibition of transient receptor potential vanilloid sub type 4 (TRPV4), a transient receptor potential ion channel, affects the severity of hearing impairment (HI). In this study, we explored the role of TRPV4 in HI using HEI-OC1 cells exposed to high glucose (HG). MATERIALS AND METHODS: HEI-OC1 cells were cultured in a HG environment (25 mM D-glucose) for 48 hours, and qRT-PCR and Western blotting were used to analyze the expression of TRPV4 at the mRNA and protein level. TRPV4 agonist (GSK1016790A) or antagonist (HC-067047) in cultured HEI-OC1 cells was used to obtain abnormal TRPV4 expression. Functional TRPV4 activity was assessed in cultured HEI-OC1 cells using the MTT assay and a cell death detection ELISA. RESULTS: TRPV4 agonists exerted protective effects against HG-induced HI, as evidenced by increased MTT levels and inhibition of apoptosis in HEI-OC1 cells. TRPV4 overexpression significantly increased protein levels of phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK), while TRPV4 antagonists had the opposite effect. Our results indicated that TRPV4 is a hyperglycemia-related factor that can inhibit cell proliferation and promote cell apoptosis by activating the MAPK signaling pathway in HEI-OC1 cells. CONCLUSION: Our results show that the overexpression of TRPV4 can attenuate cell death in HEI-OC1 cells exposed to HG.

Management of pregnancy in patients with complex congenital heart disease / 中国医师杂志

In recent years,with the continuous improvement of the diagnosis and treatment of congenital heart disease,most women with congenital heart disease can reach the childbearing age.For women with complex congenital heart disease,pregnancy can seriously endanger the safety of mother and baby.Therefore,pre-pregnancy assessment and standardized obstetric management of such patients are essential.

Effects of different doses of dexmedetomidine on acute brain edema in mice with traumatic brain injury / 临床麻醉学杂志

Objective To explore the effects of different doses of dexmedetomidine on acute brain edema in mice with traumatic brain injury (TBI).Methods A total of 132 male C57BL/6J mice were randomly divided into six groups:control group (group C),sham-operation group (group Sham),traumatic brain injury group (group TBI),Dex 20 μg/kg (group D20),40 μg/kg (group D40),and 60 μg/kg (group D60),n=22 in each group.The TBI animal model was established by electric controlled cortical impactor (eCCI),then intraperitoneal injected by the administration of different doses of dexmedetomidine at 0,2 and 4 h after TBI.Twenty-four hours post-TBI,brain water content was measured by the dry-wet method,histological observation was performed using HE staining,and aquaporin 4 (AQP4) and NF-κB expression were detected using Western blot assay,respectively.Then,the modified neurological scale scores (mNSS) on 1,2,3,and 7 d and Morris water maze (MWM) test on 4,5,6 and 7 d post-TBI were used to evaluate the neurologic deficit of TBI mice.Results After traumatic brain injury,the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB increased significantly in group TBI (P＜0.01).Different doses of dexmedetomidine significantly reduced the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB (P ＜ 0.05 or P ＜ 0.01).And meanwhile dexrnedetomidine can lessen neuronal degeneration,and inflammation response.Additionally,the effect was remarkably in group D60 compared with group D20 (P ＜ 0.05 or P ＜ 0.01).Conclusion Dexmedetomidine can lessen brain edema and cognition impairment induced with traumatic brain injury,which is a dose-effect relationship within 20-60 μg/kg,and this effect may be related to the downregulation of AQP4 and NF-κB expression.

Construction and characterization of recombinant pseudovirus particles carrying specific genomic RNA of St.Louis encephalitis virus / 军事医学

Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.

Therapeutic effects of the integrated acupuncture and Chinese herbal medicine on reflux esophagitis / 中国针灸

<p><b>OBJECTIVE</b>To compare the differences in the clinical therapeutic effects on reflux esophagitis among the combined therapy ofdecoction (the decoction for resolving the turbid, detoxification and reducing the pathologic upwardin short) and acupuncture, omeprazole and Chinese herbal medicine.</p><p><b>METHODS</b>Ninety patients were randomized into 3 groups, 4 cases of them were dropped off. Finally, there were 29 cases in the combined therapy group with acupuncture and the decoction, 29 cases in the western medication group and 28 cases in the Chinese herbal medicine group in the statistical analysis. In the combined therapy group with acupuncture and the decoction, the decoction was prescribed recurrence rate. The therapeutic effects are better than the simple application of either Chinese herbal medicine or omeprazole. for oral administration. Additionally, acupuncture was applied to Neiguan (PC 6), Zusanli (ST 36), Zhongwan (CV 12), Ganshu (BL 18), Danshu (BL 19) and Taichong (LR 3). The decoction was applied one dose a day and acupuncture was once a day. In the western medication group, omeprazole capsules, 20 mg were prescribed for oral administration, twice a day. In the Chinese herbal medicine group, the decoction was simply applied. The treatment was 8 weeks in the 3 groups and the follow-up visit was 6 months. The score of reflux disorder questionnaire (RDQ) and the changes in esophageal mucosa under gastroscope were observed before and after treatment; the clinical therapeutic effects and recurrence rate were evaluated in the 3 groups.</p><p><b>RESULTS</b>In 4 and 8 weeks of treatment, RDQ scores in the 3 groups were all reduced as compared with those before treatment (all<0.05). In 4 weeks of treatment, RDQ score in the combined therapy group with acupuncture and Chinese herbal medicine was lower than that in the western medication group (<0.05). In 8 weeks of treatment, RDQ score in the combined therapy group with acupuncture and Chinese herbal medicine was lower than those in the western medication group and the Chinese herbal medicine group (both<0.05). In follow-up visit for 6 months, the recurrence rate in the combined therapy group with acupuncture and the decoction was lower than those in the other two groups (both<0.05). In 8 weeks of treatment, the total effective rate for clinical symptoms and that observed under gastroscope in the combined therapy group with acupuncture and the decoction were all better than those in the western medication group and the Chinese herbal medicine group (all<0.05).</p><p><b>CONCLUSIONS</b>The combined therapy ofdecoction and acupuncture achieve the definite therapeutic effects on reflux esophagitis, relieve the symptoms, protect gastric mucosa and reduce the.</p>

Effects of Different Exercises on cAMP/CREB/ATF4 Pathway and Bone Formation in Bone of T2DM Mice / 中国运动医学杂志

Objective To observe the changes of bone formation in type 2 diabetes mellitus (T2DM) mice and the effect of different exercises on their cAMP/CREB/Atf4 pathway and bone formation.Methods Forty four-week-old C57BL/6 male mice were randomly divided into a normal control group (ZC) of 10 and a T2DM group of 30.T2DM was induced using the high-fat diet and injection of streptozotocin.Then,the T2DM mice were randomly divided into a T2DM control group (TC),a T2DM swimming group (TS) and a T2DM downhill running group (TD),each of 10.The TS and TD groups were trained for eight weeks as their group names indicated.Then the concentration of cAMP in serum was tested using the enzyme-linked immunosorbent assay (ELISA).The reverse transcription-polymerase chain reaction (RT-PCR) was used to test the mRNA expression of the cAMP-response element binding protein (CREB),activate transcription factor 4 (ATF4),osteocalcin (OC),bone gla-protein (OCN)and bone sialoprotein (BSP) in the left tibia,and Western blotting was employed to test the protein expression of CREB in the right femur.The bone marrow mesenchymal stem cells (BMSCs) were taken and induced to differentiate into osteoblasts (OBs) and dyed using the alkaline phosphatase (ALP) solution.The left side hindlimb bone was taken and scanned the bone mineral density (BMD) of the distal end using the Skyscan Micro-CT.Results Compared with group ZC,the concentration of cAMP declined in group TC.Moreover,the mRNA expression of CREB,ATF4,OC,OCN and BSP as well as the protein expression of CREB of group TC were significantly down-regulated(P＜0.01 or P＜0.05),together with the OB osteogenic capacity and BMD (P＜0.01) compared to group ZC.Compared with group TC,significant increase was observed in the mRNA expression of OC and OCN (P＜0.01 or P＜0.05),as well as the OB osteogenic capacity of group TS.The concentration of cAMP of group TD decreased,the mRNA expression of CREB,ATF4,OC and OCN,as well as the protein expression of CREB were all significantly up-regulated (P＜0.01 or P＜0.05) compared with group TD.The OB osteogenic capacity and BMD(P＜0.05) of group TD also increased significantly.Compared with group TS,the concentration of cAMP(P＜0.05) and the OB osteogenic capacity increased,and the mRNA expression of CREB,ATF4 and OC of group TD increased significantly(P＜0.01 or P＜0.05).Conclusion The bone formation metabolism of type 2 diabetic mice is inhibited.The downhill running is superior to swimming in promoting the osteoblast differentiation and bone formation,as well as the bone mineral density through activating the cAMP/CREB/Atf4 pathway in the bone of T2DM mice.

Mechanism of Coix Seed Extract in Improving Free Fatty Acid Metabolism in Rats with Fatty Liver / 中国药师

Objective:To observe the effect of coix seed extract on non-alcoholic fatty liver disease (NAFLD), and discuss the mechanism of coix seed extract in the treatment of NAFLD in the repect of free fatty acid. Methods:Totally 60 SD rats were randomly divided into 6 groups with 10 ones in each, the normal group, the model group, the positive group ( Xuezhikang capsules, 1 g · kg-1), three coix seed extract groups respectively at high (20 g·kg-1), medium (10 g·kg-1) and low (5 g·kg-1) dosage. The normal group was given basic diet, the other 5 groups were given high fat diet to establish NAFLD rat model, the modeling time was 8 weeks, and the drug treatment started from the 5th week till the 8th week. The liver weight and fat weight and index were observed after the drug intervention. The serum total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), free fatty acid (FFA), fatty acid synthase (FAS), acetyl coenzyme A carboxylase (ACCa-se), C two acyl coenzyme A (MALONYL-CoA) , adenosine content of protein kinase ( AMPK) and adiponectin ( ADP) activated by glycosides were detected. Results: Com-pared with those in the normal group, the lipid metabolism related indices in the model group were significantly abnormal (P<0. 05), which suggested that the model was established successfully. After the drug intervention, the lipid metabolism related indicators in the positive group and the three drug groups were adjusted in varying degrees. Compared with those in the model group, TC, FFA and AMPK expression levels increased notably in the high dosage group and the positive group (P<0. 05), and the levels of ALT, AST, FAS, ACCa-se, MALONYL-CoA decreased at the same time (P<0. 05). Coix seed extract could decrease the body weight and liver wet weight in NAFLD rats and improve the related index significantly. Conclusion: The findings indicated that coix seed extract is highly effective in improving the pharmacological effect on NAFLD induced by high-fat diet, and the mechanism is achieved through ADP-AMPK-ACCase-malony-CoA-FFA axis.

ROS mediates regulation of intracellular Ca2 +induced by angiotensin II in primarily cultured medullary neurons / 中国病理生理杂志

AIM:To investigate the role of reactive oxygen species (ROS) in the regulation of intracellular Ca2+induced by angiotensin II ( Ang II) in the primarily cultured medullary neurons .METHODS:Primarily cultured me-dullary neurons were prepared from 14-day-old embryos of Sprague-Dawley rats in the study .The identification of medullary neurons was assessed by double-labeling immunofluorescence .To explore the role of ROS , mainly the superoxide ( O2 ·-) , the O2 ·-generation was measured using the fluorogenic probe dihydroethidium ( DHE) .To determine intracellular free cal-cium concentration ( [ Ca2+] i ) , the neurons were loaded with the Ca 2+-specific dye Fura-2/AM.The cell viability after adding Ang II was also examined using CCK-8 assay.RESULTS:Most of the cultured cells were medullary neurons , more than 80%of which were glutamate positive neurons .Ang II (5 μmol/L) increased the level of ROS within 10 min in the medullary neurons .Ang II at 5μmol/L induced a significant [ Ca2+] i increase in the medullary neurons , and the effect of Ang II occurred rapidly and reached a peak within 20 min after administration.The level of [Ca2+]i started to decline after washout .The Ca2+elevation induced by Ang II was significantly decreased by apocynin or TEMPOL .No significant differ-ence in the cell viability between control group and 5μmol/L Ang II treatment group was observed .CONCLUSION:ROS is involved in the regulation of [Ca2+]i induced by Ang II in the primarily cultured medullary neurons , suggesting a poten-tial intracellular signaling mechanism involved in the Ang II-mediated oxidant regulation of central neural control of blood pressure.

Comparison of Sirolimus and Everolimus Drug-eluting Stents for Treating the Patients With Non ST-elevation Acute Coronary Syndrome / 中国循环杂志

Objective: To compare the efifcacy and safety of sirolimus-eluting stent (SES) and everolimus-eluting stent (EES) for treating the patients with non-ST segment elevation acute coronary syndrome (NSTE-ACS). Methods: A total of 400 NSTE-ACS patients treated in Jining Medical College Hospital from 2013-09 to 2014-09 were studied. According to different stents, the patients were divided into 2 groups: SES group,n=220 and EES group,n=180. A prospective follow-up study was conducted for 1.5 years to compare the incidence rate of major adverse cardiovascular events (MACE). The patients were further stratiifed by GRACE scores as Low risk group (score140). MACE free survival was studied by Kaplan-Meier curve and analyzed by Long-rank test, predictive value of GRACE for 1.5 year MACE incidence rate was examined. Results: There were 355/400 (89%) patients completed (16.7 ± 5.7) months of follow-up study including 205 in SES group and 150 in EES group. MACE occurrence rates were similar between SES group and EES group (16.10% vs 18.0%), P>0.05. By GRACE score stratiifcation, MACE rates in High risk SES group were higher than High risk EES group (48.00%vs 16.00%),P0.05. ROC curve indicated that the predictive value of GRACE score for 1.5 year MACE incidence was for AUC=0.762, 95% CI (1.026-1.050),P<0.001. Conclusion: Implanting of EES would be more beneifcial for NSTE-ACS patients with high GRACE risk; GRACE score has the better predictive value for their long-term prognosis.

Extraction Technology for Antitumor Active Components in Fructus Ligustri Lucidi / 中国药师

Objective:To optimize the extraction process of antitumor active ingredients in fructus ligustri lucidi. Methods:Using fructus ligustri lucidi medicinal herb as the raw material, the extraction process of luteolin and pelargidenon in the herb was screened by D-optimal design. The extraction yield of different extraction processes was compared to obtain the optimal extraction conditions. Re-sults:The best extraction conditions were as follows:adding 12-fold 95% ethanol, extracting 2 times with 60 minutes for each time. Conclusion:The optimal extraction technology can provide significant reference for the further pharmacodynamic effect development of fructus ligustri lucidi.

Study on in vitro Antimicrobial Activity of Humulus Water Extract / 中国药师

Objective:To study the in vitro bacteriostasis of water extract of humulus. Methods:By using double dilution method and 96-well plates trace broth dilution method, the inhibitory test of water extract of humulus was conducted on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The minimum inhibitory concentration ( MIC ) , minimum bactericidal concentration ( MBC) and the diameter of inhibition zone were determined. Results: The drug showed different sensitive degree on the test strains. MIC and MBC for Escherichia coli were 31. 25 mg·ml-1 , that for Staphylococcus aureus were 3. 91 mg·ml-1 , and that for Pseudo-monas aeruginosa was 125 and 250 mg·ml-1 . Conclusion:With the water extract of humulus as the research object, the study on the in vitro bacteriostasis effect of humulus scandens is closer to the clinical application form, which provides reference for the further re-search on the related preparation of humulus scandens.

Construction of human Ewing sarcoma cell line with knockdown of inhibitor of differentiation 2 gene / 肿瘤研究与临床

Objective To construct short hairpin RNA (shRNA) expression plasmids against the inhibitor of differentiation 2 (Id2) gene and establish a suitable cell model for the role of Id2 in proliferation and differentiation of human Ewing sarcoma cell.Methods Three shRNA sequences targeting Id2 gene were designed and inserted into the pGPU6/GFP/Neo (-shRNA-Id2) expression vectors.The recombinant pGPU6/GFP/Neo-shRNA-Id2 plasmids were introduced into Ewing sarcoma RD-ES cells by liposome-mediated transfection.The knock-down efficiency of Id2 in infected RD-ES cells was verified by Western blot assay.The cell growth and cell cycle changes were evaluated by cell counting and flow cytometry between transfected cells and control cells.Results The Id2 expression decreased 54 ％ and 57 ％,respectively,in RD-ES cell line which were transfected with the shRNA-Id2-543 and shRNA-Id2-593 plasmids compared with the control group cells by Western blot analysis.The cell growth assay demonstrated that the cell number in transfected cells was significantly decreased during 6-7 d compared with the control group (P ＜ 0.05).The cells at the S-phase of cell cycle were increased [(36.60±1.53) ％ and (44.89E2.46) ％ vs (29.73±2.03) ％,P ＜ 0.05],and no significant changes at the G2 phase or even reduction in the transfected cells.Conclusions Id2 stable knock-down cell lines are successfully established.The reduced expression of Id2 is related with decreased cell growth and cell cycle arrest in the S-phase.Id2 maybe plays an important role in proliferation of Ewing sarcoma cell.