Objective To evaluate the molluscicidal effect of black plastic mulching in mountainous areas of Yunnan Prov-ince. Methods In Leqiu Village,Nanjian County,Yunnan Province,the terraced fields with Oncomelania hupensis snails were divided into A,B,C,D,E groups,and of which,A,B,C,D groups,as the experimental groups,were administered with sim-ple black plastic mulching for 30 days,60 days,90 days and 120 days respectively;Group E,as the control group,was adminis-tered with 50%niclosamide ethanolamine salt wettable powder spraying. Results In Group C and D(simple black film mulching for 90 days and 120 days),no snails were found after the administration,the average density of living snails decreased by 100%, and the cost of one year was similar to that of the control group. Conclusion The molluscicidal effect of black plastic mulching is good in mountainous areas.

Objective:To study the interaction of the inflammatory cytokines expression between CD4+ T cells and primary human umbilical vein endothelial cells (HUVECs) infected by dengue virus (DENV-2).Methods:PBMC was extracted from white blood cells by density gradient centrifugation,CD4+ T cells were sorted by immunomagnetic beads.The expression of CD3 and CD4 molecules on the surface of cells was detected by flow cytometry to identify the purity of CD4+ T cells.First,HUVECs were pretreated by specific-S1 P1 receptor agonist CYM-5442 for 24 h,second,infected by DENV-2 on the titer of 103 TCID50,then co-culturing with CD4+ T cells,The relative expression of NS1 partial sequence and IL-6,L-8 mRNA of HUVECs,and IL-4,IL-17,TNF-α,IFN-γ of CD4+ T cells detected by Real-time RT-PCR.IL-6 and IL-8 secreted in cultured supematant analyzed by ELISA.Results:The purity of CD4+ T cells was (98.02±0.32) ％.The expression of NS1 gradually increased to 24 h (3.03±0.26,P＜0.001),decreased after reaching the peak.The relative expression of NS1 in the group of co-cultured with CD4+ T cells was lower than other groups.After infection,the expression of IL-6 and IL-8 were up-regulated,and the expression of IL-6 and IL-8 at each time point was significantly increased after co-culturing with CD4+ T (P＜0.01).IL-6 of CYM-5442 pretreatment group,in 24 h (28.91±2.34,P＜0.05),36 h(19.36±0.1,P＜ 0.05) and 72 h(13.84±0.82,P＜0.05) was significantly decreased,the expression of IL-8 also decreased significantly.The mRNA expression of IL-4,IL-17,TNF-α and IFN-γ in CD4+ T cells was significantly increased after co-culturing with HUVECs.After the treatment with CYM-5442 group,the expression was decreased.Conclusion:DENV-2 could infect the primary HUVECs,and the expression of NS1 was inhibited after co-culturing with CD4+ T cells.CD4+ T cells can not only enhance the activation of HUVECs infected by DENV-2,but also can be activated by the infected HUVECs infected with DENV-2.

Objective To explore the effects of different period of insulin-like growth factor-1(IGF-1)on B cell lymphoma/leukeamia-2 gene(BCL-2)and BCL-2 associated X protein(BAX)of myocard in mice with viral myocarditis.Methods Male Balb/c mice were randomly divided into control group,coxsackie viral B3(CVB3)infection group(infection group),IGF-1-treated group in earlier stage(0-6 d)and acute stage(7-13 d),injected IGF-1 of exogenous in abdominal cavity [30 ?g/(kg?d)].Animals were killed at the 14 day.The immunohistochemical studies and medical imagine analysis system were performed to evaluate the expression of BCL-2 and BAX in myocardial tissue.Results In infection group,the expression of BCL-2 reduced and BAX increased than those of control and treatment group(Pa

AIM:To observe the effects of adipose differentiation-related protein ( adipophilin) on the expres-sion of inflammatory factors in RAW264.7 macrophage and to clarify the related mechanism.METHODS:The cell models with high expression and low expression of adipophilin were constructed by transfecting PA317 packaging cells with stable high or low expression adipophilin retroviral vectors into the RAW264.7 cells.The concentrations of IL-6, MCP-1 and TNF-αin the cell culture medium were detected by ELISA.The protein levels of AP-1, p-AP-1, ERK1/2 and p-ERK1/2 were measured by Western blot.The protein levels of adipophilin, p-ERK1/2 and p-AP-1 and the releases of the inflamma-tory factors in the RAW264.7 cells treated with or without ERK1/2 inhibitor PD98059 or AP-1 inhibitor curcumin were de-termined.RESULTS:The RAW264.7 cells with high expression of adipophilin had higher levels of IL-6, MCP-1 and TNF-α, and higher protein levels of p-AP-1 and p-ERK1/2 than those in the cells with low expression of adipophilin. ERK1/2 inhibitor had no significant effect on the expression of adipophilin, but the protein expression of ERK1/2 and AP-1 was significantly inhibited (P<0.05).The administration of AP-1 inhibitor curcumin had no significant effect on the protein expression of adipophilin and ERK1/2, but the protein expression of AP-1 was significantly inhibited (P<0.05). At the same time, the releases of inflammatory factors IL-6, MCP-1 and TNF-αwere significantly decreased.CONCLU-SION:Adipophilin may regulate the expression of inflammatory factors through ERK1/2-AP-1 pathway in RAW264.7 mac-rophages.

AIM:To study the effects and mechanisms of ethanol on chloride channels in poorly differentiated nasopharyngeal carcinoma CNE-2Z cells.METHODS:The effect of ethanol on the cell growth was analyzed by MTT as-say.The technique of whole-cell patch-clamp was used to detect the chloride current .The characteristics of the chloride current were analyzed by using the chloride channel blockers .The siRNA technique was used to analyze the molecular basis of the ethanol-sensitive chloride channels .RESULTS: Under isotonic conditions , the background current was weak and stable.Ethanol at concentrations of 0.17~170 mmol/L activated a chloride current in a concentration-dependent manner (an inverted U-shape), with a maximum effect at the concentration of 17 mmol/L.The currents showed obviously outward rectification and were susceptible to extracellular hypertonicity and the chloride channel blocker , 5-nitro-2-(3-phenylpropyl-amino) benzoic acid ( NPPB) .ClC-3 siRNA obviously decreased the currents activated by ethanol .CONCLUSION:Ex-tracellular ethanol induces chloride currents through activating the ClC-3 chloride channels .

Objective To investigate the molecular mechanism of retinoic acid in targeted treatment of craniopharyngioma by detecting the expression of RARαand PPARβ/δin craniopharyngioma cells and analyzing the correlation between the expression and effect of retinoic acid. Methods The expression of RARα and PPARβ/δ in craniopharyngioma cells from 31 patients cultured in vitro was quantified by reverse transcription-PCR. The inhibition rates of RA on craniopharyngioma with different expression of RARα and PPARβ/δ were detected by using MTT assay, and the correlation between the expression of RARα and PPARβ/δand the effect of RA was analyzed. Results 1. The RT-PCR results showed that the expression levels of PPARβ/δand RARα mRNA were different. Craniopharyngioma cells from 31 patients in primary culture were divided into three groups according the expression levels of nuclear receptor: PPARβ/δ>RARα group, RARα>PPARβ/δ group and RARα>>PPARβ/δ group. 2.MTT results showed that the inhibition rate of RARα>>PPARβ/δgroup was significantly higher than the other groups under same drug, the differences had statistical significance ( <0.01) . Conclusions The expression of PPARβ/δ, RARα can be used to evaluate the effect of RA in treatment of craniopharyngioma. The craniopharyngioma with low-expression of PPARβ/δ is more sensitive to RA. Targeting higher RARα or targeting lower PPARβ/δ is beneficial to the treatment of craniopharyngiomas.

Objective To explore the effects of HAVCR2 siRNA on apoptosis and cell cycle in the radiation-caused genomic instable liver cells.Methods RNAi was used to inhibit HAVCR2 gene transcription.Cellular apoptosis and cell cycle distribution were detected by flow cytometry (FCM).Expression of p53 gene was assayed by real time fluorescence quantitative PCR.Results HAVCR2 gene was effectively inhibited by RNAi (t =19.21,P ＜ 0.05).After siRNA transferring,cell apoptosis (t =3.65,P ＜ 0.05) and p53 gene expression (t =4.82,P ＜ 0.05) were decreased,and G2-phase arrest was induced(t =-3.41,P ＜ 0.05).Conclusion HAVCR2 siRNA can decrease the generation of apoptosis in the genomic instable liver cells and blocks the cells at G2 phase.

Objective To perform digital 3D measurement of anatomic parameters associated with vertebral units fixed by ribs and pedicle fixation of T4-T10,providing evidence for clinical application.Methods Spiral CT scan of T4-T10 vertebrae was conducted in 15 normal adults without spinal disorder.The data were imported into computer to establish units and disc models of thoracic and rib vertebrae T4-T10 using Mimics 16.0 software and Ansys 11.0 finite element software.The following data were measured in the vertebral pedicle-rib unit fixation group (group A) by software:transverse diameter of vertebral pedicle-rib unit,length of the outside pedicle screw,inclination angle of pedicle screw,and maximum and minimum inclination angles of pedicle screw;the following data were also measured in the pedicle fixation group (group B) by software:pedicle transverse diameter,length of pedicle screw,inclination angle of pedicle screw,and maximum and minimum inclination angles of pedicle screw.The corresponding parameters were compared between groups A and B.Results In both groups,the transverse diameters and screw lengths gradually increased with the increase in the vertebral sequence while the inclination angle of pedicle screw,and maximum and minimum inclination angles of pedicle screw decreased with the increase in the vertebral sequence.All the parameters in groups A were significantly larger than the corresponding ones in group B (P ＜ 0.05).Conclusions Of the spinal segment of T4-T10,the rib vertebral unit fixation can provide greater safe screw angles and screw diameters for clinical surgery.This is of vital significance for reducing the surgical complexity and improving screw prehension.

Objective To analyze the differences of the clinical and neuropathological features among the common Charcot-Marie-Tooth disease (CMT) subtypes.Methods There were 81 CMT patients confirmed by genetic testing from 2005 to 2015 in Department of Neurology,Peking University First Hospital,including 31 cases of CMT1A (38.3％),19 cases of CMTX1 (23.5％),16 cases of CMT2A2 (19.8％) and 15 cases of 9 rare types of CMT (1.2％-4.9％).We compared the onset age,duration,muscles weakness of legs,frequency of pes cavus,and main pathological changes of the sural nerve biopsy in 48 cases of the common CMT subtypes.Results The mean age of the onset was (12.00 ± 6.77) years in CMT1A patients,(11.81 ±4.65) years in CMTX1 patients and (5.00 ±2.68) years in CMT2A2 patients (Brown-Forsythe test,P =0.001).The duration was (12.00 ± 6.75) years in CMT1A patients,(8.50 ± 4.75) years in CMTX1 patients and (5.00 ± 2.73) years in CMT2A2 patients (Brown-Forsythe test,P =0.001).The muscle force of the dorsi flexors was Ⅳ (0,Ⅴ) in CMT1A patients,Ⅲ + (0,Ⅳ) in CMTX1 patients and 0 (0,Ⅳ) in CMT2A2 patients (H =11.359,P =0.020).The pes cavus appeared in 15/23 cases of CMT1A,10/16 cases of CMTX1 and 1/9 cases of CMT2A2 (Fisher test,P=0.017).The leukoencephalopathy appeared only in 3 cases of CMTX1 and the visual loss appeared only in 3 cases of CMT2A2.The onion-bulb formations of myelinated fibers appeared in 23/23 cases of CMT1 A,5/16 cases of CMTX1 and 2/9 cases of CMT2A2(Fisher test,P =0.000).The axonal regeneration appeared in 16/23 cases of CMT1A,16/16 cases of CMTX1 and 9/9 cases of CMT2A2 (x2 =7.666,P =0.016).There were significant differences among the three common CMT subtypes in the above parameters.Conclusions CMT1A,CMT2A2 and CMTX1 are the most common subtypes of CMT in the present study.For the clinical diagnosis,more attention should be paid to the onset of the disease,duration,muscles weakness,pes cavus,cerebral symptoms and visual loss.The present frequency of onion-bulb and the axonal regeneration of myelinated fibers help the different pathological diagnosis among them.

Objective To report the clinical and peripheral neuropathological findings in two patients with autosomal recessive Charcot-Marie-Tooth disease 2K(AR-CMT2K).Methods Case one was a nine year-old girl.She had distal weakness of lower limbs for six years, with calf atrophy and contracture of Achilles tendon for three years.Case two was an eight year-old boy.He had distal weakness of lower limbs with contracture of Achilles tendon and calf muscle atrophy for three years, and proximal weakness of low limbs for two years.The motor nerve conduction velocities in median nerves were 48.1 m/s in case one and 47.6 m/s in case two.The compound motor action potential amplitude of median nerves decreased by 46% in case one and 69% in case two.Sural nerve biopsies and gene targeted next-generation sequencing were performed in both patients.Results Density of myelinated fibers was 8 407/mm2 in case one and 7 714/mm2 in case two.The ratio of myelinated fibers with diameter over 8 μm was 2.6% in case one and 0 in case two.Both patients had small regenerating cluster of myelinated fibers.Thin myelinated fibers appeared in case one.In case two, atypical onion bulb formations with focal folded myelin appeared, and electromicroscopy revealed mitochondrial aggregate in axons.Compound heterozygous mutations of ganglioside-induced differentiation associated protein 1 gene were detected in both patients, including c.767A>G(p.H256R) and c.466G>A (p.A156T) in case one and c.767A>G and 845G>A(p.R282H) in case two.Conclusions Contracture of Achilles tendon may appear in early childhood of AR-CMT2K patients.The main pathological changes in sural nerve are loss of large myelinated fibers, mitochondrial aggregate in axons and myelin abnormalities.