BACKGROUND:Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia,excessive release of inflammatory factors and redox imbalance.However,the specific mechanism is not fully understood. OBJECTIVE:To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS:Passage 3 BV2 microglia were divided into three groups:control group was cultured for 24 hours(without any treatment).Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours.Lipopolysaccharide+nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours.After the processing,cell proliferation was detected by CCK-8 assay.The level of intracellular reactive oxygen species was detected by fluorescent probe.The mRNA expression levels of nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by qRT-PCR.The protein expression levels of nuclear factor κB p65,p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of lipopolysaccharide group was decreased(P<0.001).Compared with the lipopolysaccharide group,the cell proliferation activity of lipopolysaccharide+nobiletin group was increased(P<0.001).(2)Compared with the control group,the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide+nobiletin group(P<0.01).(3)Compared with the control group,the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group(P<0.001,P<0.01).Compared with the lipopolysaccharide group,mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide+nobiletin group(P<0.01,P<0.05).(4)Compared with the control group,the protein expression levels of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β in were increased the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the expression of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β was decreased in the lipopolysaccharide+nobiletin group(P<0.001).(5)These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway.

BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment. OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism. METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1β group,sinomenine+interleukin-1β group,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1β group,and sinomenine+zinc protoporphyrin+interleukin-1β group.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected. RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an"S"curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the"platform stage"where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80 μmol/L(P>0.05).Interleukin-1β significantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

BACKGROUND:Intervertebral disc degeneration is caused by damage and degeneration of the nucleus pulposus and annulus fibrosus tissues inside the intervertebral disc,resulting in structural and functional changes of the intervertebral disc.However,there is yet no effective drug treatment for intervertebral disc degeneration. OBJECTIVE:To investigate the inhibitory effect of syringin on intervertebral disc degeneration. METHODS:A total of 10 male Sprague-Dawley rats were selected,and the coccygeal intervertebral disc(Co4/Co5)of each rat was set as model group,Co5/Co6 intervertebral disc as syringin group,and Co6/Co7 intervertebral disc as control group.The control group did not receive any treatment.In the model group and syringin group,a miniature puncture needle was used to puncture the annulus fibrosus to establish an intervertebral disc degeneration model.Immediately after modeling,2.5 μL of normal saline and syringin solution(5 μmol/L)were given in the model and syringin groups,respectively.Four weeks after injection,the samples were taken.The degree of intervertebral disc degeneration in rats was observed by hematoxylin-eosin and safranine O-fast green staining.The expressions of type Ⅱ collagen,aggrecan and matrix metalloproteinases 3 and 13 in intervertebral disc tissue were analyzed by immunohistochemical staining. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that in the model group,the height of intervertebral disc decreased,the cartilage endplate became thinner and cracked,the fibrous ring structure was disordered and cracked,and the nucleus pulposus disappeared;in the syringin group,the height of intervertebral disc was normal or slightly lower than that in the control group,the degree of cartilage endplate degeneration was lighter than that in the model group,the fiber circle permutation was relatively regular with no cracks,and the nucleus pulposus was partially shrunk.Safranine O-fast green staining showed that in the model group,the cartilage endplate of the intervertebral disc was defective and the calcified layer of cartilage became thinner,showing obvious degeneration.The structure and morphology of intervertebral disc cartilage endplate in the syringin group recovered to some extent.Immunohistochemical staining showed that,compared with the control group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage were decreased in the model group(P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 increased(P<0.000 1).Compared with the model group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage tissue were increased in the syringin group(P<0.001,P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 decreased(P<0.001,P<0.000 1).These results showed that syringin could improve the structure and function of intervertebral disc by inhibiting the expression of matrix metalloproteinases 3 and 13 and increasing the expression of type Ⅱ collagen and aggrecan,thus preventing and slowing down the procession of intervertebral disc degeneration.

Animals ; Carrier Proteins/metabolism* ; Histidine/metabolism* ; Humans ; Methylation ; Methyltransferases/metabolism* ; Neoplasm Proteins/metabolism* ; Neoplasms/metabolism* ; Protein Processing, Post-Translational

Objective To compare the effects of different anesthetics on the recovery of neurologi-cal function after intracranial aneurysm embolization. Methods One hundred patients of both sexes with aneurysmal subarachnoid hemorrhage, aged more than 18 yr, with body mass index of 18. 5-24. 0 kg∕m2 , of American Society of Anesthesiologists physical status Ⅱ or Ⅲ and WFNS grade Ⅰ-Ⅳ, with the thick-ness of subarachnoid hemorrhage more than 4 cm, were divided into 2 groups (n= 50 each) using a random number table: propofol group (group P) and sevoflurane group (group S). After anesthesia induction, group P received intravenous infusion of propofol 100-300 μg·kg-1 ·min-1 , while the end-tidal sevoflu-rane concentration was maintained at 1. 4％-3. 5％ in group S. Immediately before induction (T0 ), imme-diately after the end of induction (T1 ), immediately after successful embolization of aneurysm (T2 ) and at 1, 2, 3 and 5 days after surgery (T3-6 ), central venous blood samples were collected for determination of plasma neuron-specific enolase and S100β protein concentrations by enzyme-linked immunosorbent assay. The development of postoperative cerebral vasospasm and delayed ischemic neurological deficit was recorded. The patients were followed up, and the Glasgow outcome score and occurrence of newly developed cerebral infarction were recorded within 6 months after surgery. Results There was no significant difference in the concentrations of plasma neuron-specific enolase and S100β protein at each time point, incidence of postop-erative cerebral vasospasm and delayed ischemic neurological deficit, or Glasgow outcome score and inci-dence of newly developed cerebral infarction within 6 months after surgery between two groups (P>0. 05). Conclusion Propofol and sevoflurane exert no effect on the recovery of neurological function after intracra-nial aneurysm embolization.

Research teams work together to solve the technical bottleneck in the Panax notoginseng industry.These research directions mainly focus on breeding of new variety,alleviating the continuous cropping obstacles,building standard system and so on.We used methods such as biotechnology,information technology and so on,to realize digitization,network and intellectualization in the process of selection of producing areas,field management,harvest storage,solved key issues of the precise selection of producing areas,variety breeding,soil improvement,fertilization management,and realized the precision and optimization of ginseng cultivation.We built the composite technology system of pollution-free notoginseng.This technology system mainly contained the breeding of new cultivar,the composite technology of soil improvement,the cultivation system of pollution-free notoginseng and quality traceability system.A new disease-resistant cultivar of notoginseng was bred,which offered feasibility for the notoginseng continuous cropping.We first presented composite improvement technology of notoginseng continuous cropping soil.This improvement technology has been used in the field.This technology system of soil restoration and improvement guaranteed notoginseng continuous cropping.A first standard system of pollution-free notoginseng was issued.It established standard of high-quality notoginseng materials.Finally,quality traceability system of notoginseng was finished,which safeguarded the controllable quality.This composite technology system offers references for the cultivation of other Chinese herbal medicines.

Objective To evaluate clinical efficacy of tigecycline alone or in combination with other antimicrobials in treating infection caused by carbapenem-resistant Acinetobacter baumannii (CRAB).Methods Patients with hos-pital-acquired pneumonia (HAP)and/or bloodstream infection in the intensive care unit of a hospital between Janu-ary 2013 and June 2014 were selected,efficacy of tigecycline treatment was analyzed.Results Of 25 patients with CRAB infection,21 were with HAP,2 with bloodstream infection,and 2 with both HAP and bloodstream infec-tion.13 cases were multidrug-resistant Acinetobacter baumannii (MDRAB), 10 cases were extensively drug-resistant Acinetobacter baumannii (XDRAB).The susceptibility rate of 25 CRAB isolates to tigecycline was 84%. After treated with tigecycline,white blood cell count(WBC),C-reactive protein (CRP),and procalcitonin (PCT) all significantly decreased (all P<0.01).The clinical effective rate,bacterial clearance rate,and 30-day mortality were 68.00% (17/25),66.67%(14/21),and 28.00% (7/25)respectively;effective rate of 21 cases of HAP was 76.19% (16/21),1 case of bloodstream infection was effective,2 cases of HAP combined bloodstream infection died.Conclusion Tigecycline is effective in the treatment of HAP caused by CRAB,but the therapeutic effect on bloodstream remains uncertain,further research is needed.

AIM:To investigate the regulation mechanism for insufficient KChIP 2 expression induces Ito,f downregulation and arrhythmogene-sis in cardiac hypertrophy .METHODS:Bidirectional manipulations of MG 53 expression were performed by adenoviral overexpression of MG53 or knockdown of MG53 with RNA interference in neonatal rat ventricular myocytes with or without PE stimulation .Ito,f was re-corded with patch clamp in whole-cell mode 48 h after adenoviral transfection .Then the WT or MG53 knockout ( MG53 -/-) mouse model of left ventricular hypertrophy induced by transverse aortic constriction ( TAC) were used to detect the susceptibility to ventricu-lar arrhythmia.RESULTS: Here, we show muscle-specific MG53 regulates KChIP2 expression and Ito,f densities, where they are downregulated in hearts from MG53 knockout mice and MG53 knockdown rat cardiomyocytes , but upregulated in MG53 overexpressed cells.MG53 expression is decreased in phenylephrine ( PE)-induced cardiomyocyte hypertrophy and restoration of MG 53 rescues PE-induced downregulation of KChIP2 and Ito,f.Furthermore, MG53 is decreased in a mouse model of hypertrophy induced by transverse aortic constriction and ablation of MG 53 increases the susceptibility to ventricular arrhythmia by exaggerating Ito,f remodeling.CON-CLUSION:These findings establish MG53 as a novel regulator of Ito,f and its central role in arrhythmogenesis in hypertrophy .

OBJECTIVETo investigate the distribution patterns of mosquito and mosquito-borne viruses in Dehong prefecture, Yunnan province, China.

METHODSMosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July, 2007 and 2010. Mosquito were cell cultured for viral isolation, and positive isolates were identified using RT-PCR and sequence analysis.

RESULTSA total of 43 634 mosquito comprised of 29 species representing six genera were collected. Culex tritaeniorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeniorhynchus, identified as genotype I Japanese encephalitis virus (JEV). One strain was identified from Cx. tritaeniorhynchus, as Getah virus (GETV). Two strains isolated from Cx. tritaeniorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus (CppDNV).

CONCLUSIONCx. tritaeniorhynchus had been the major species of mosquito and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. Genotype I JEV, GETV and CppDNV were the vectors causing transmission of mosquito-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

Alphavirus ; isolation & purification ; Animals ; Arboviruses ; classification ; isolation & purification ; China ; Culicidae ; virology ; Disease Vectors ; classification ; Encephalitis Virus, Japanese ; isolation & purification

Objective To evaluate the treatment effects of the olecranon comminuted fracture by the angle stability prin-ciple combined with support under the articular surface. Methods From November 2008 to June 2012, 13 patients (7 male and 6 female, aged from 20 to 77 years) suffered from olecranon comminuted fracture were treated with Synthes anatomy locking com-pression plate and screws combined with Kirschner wires supporting under the articular surface through the posterior median ap-proach. Fractures occurred in the left elbow in 9 and in the right elbow in 4. According to the Mayo classification, 11 cases were classified as Mayo type IIB and 2 as IIIB. According to the Schatzker-Schmeling classification, 2 fractures were type A2, 9 type C, and 2 type D. One patient with type C fracture also had type I coronoid fracture according to Regan and Morrey classification. The Mayo elbow performance index (MEPI) and the shortened disabilities of the arm shoulder and hand (Quick-DASH) were employed to evaluate the functions. All patients took the satisfaction survey and X-ray during the follow-up. Results All patients were fol-lowed-up for 8 to 41 months. The mean ROM of the elbow joint was 112° (range, 65°-140° ), and the mean rotation angle of the forearm was 170° (range, 150°-180°). The mean score for the MEPI was 96 (range, 85-100), excellent 12, good 1. The mean score for the Quick-DASH was 6.2 (range, 0-16.7). In the satisfaction survey, 8 patients were very satisfied, 4 satisfied, 1 common level. All patients took the X-ray during the follow-up and all of them had achieved fracture union completely with the mean time period as 12.7 weeks (range, 11-24 weeks). No cubitus varus, valgus, and instability were found in all patients. No complication, such as infection, ulnar nerve injuries and etc. was found. Five patients complained about mild discomfort caused by internal fixation at 3 months after surgery. Three of them were diagnosed as joint stiffness because of ROM within 100° . One case in 3 elbow stiffness patients developed osteoarthritis after eight months, but no pain during joint activity. Internal fixation was removed in one case;an-other young patient took the operation for removal of internal fixation and release of medial ligament;the third patient refused oper-ation. Conclusion Angle stability principle (Synthes anatomy locking compression plate and screws) combined with supporting under the articular surface (Kirschner wires) can achieve stable fixation in treating olecranon comminuted fracture. It may realize the early exercise of the elbow joint with excellent clinical results. Therefore, it may be an optional choice in clinical practice.