Objective To investigate the effect of type 2 dengue virus(DENV-2)infection on autophagy in human umbilical vein endothelial cells(HUVECs)and the mechanism mediating the inhibitory effect of baicalin against DENV-2 infection.Methods Cultured HUVECs with DENV-2 infection were treated with different concentrations of baicalin,and the changes in autophagy of the cells were detected using transmission electron microscopy.Lyso Tracker Red staining was used to examine pH changes in the lysosomes of the cells,and the expressions of ATG5,beclin-1,LC3,P62,STX17,SNAP29,VAMP8,and PI3K/AKT signaling pathway-related proteins were detected by Western blotting.DENV-2 replication in the cells were evaluated using RT-qPCR.The differentially expressed proteins in DENV-2-infected HUVECs were identified by proteomics screening.Results Treatment with baicalin did not significantly affect the viability of cultured HUVECs.Proteomic studies suggested that the PI3K-AKT pathway played an important role in mediating cell injury induced by DENV-2 infection.The results of RT-qPCR demonstrated that baicalin dose-dependently inhibited DENV-2 replication in HUVECs and produced the strongest inhibitory effect at the concentration of 50 μg/mL.Transmission electron microscopy,Lyso Tracker Red staining,RT-qPCR,and Western blotting all showed significant inhibitory effect of baicalin on DENV-2-induced autophagy in HUVECs.DENV-2 infection of HUVECs caused increased cellular expressions of LC3 and P62 proteins,which were significantly lowered by treatment with LY294002(a PI3K inhibitor).Conclusion Baicalin inhibits DENV-2 replication in HUVECs and suppresses DENV-2-induced cell autophagy by inhibiting the PI3K/AKT signaling pathway.

[Objective]To analyse and summarize Professor CHEN Mingxian's clinical experience in the treatment of gastric cancer by"tonifying the spleen and stomach,removing stasis poison,clearing cancer poison".[Methods]Through clinical learning and medical case analysis,this paper elucidates Professor CHEN's understanding of the etiology and pathogenesis of gastric cancer,gives examples of the treatment of gastric cancer,and attaches two cases.[Results]Professor CHEN thinks weakness of the spleen and stomach is a prerequisite for the occurrence of gastric cancer,which runs through the occurrence and development of gastric cancer.Stasis coagulation is the key link for the transformation of precancerous lesions into gastric cancer,and the residual cancer poison is the main pathogenesis of the recurrence and metastasis of gastric cancer.In the treatment,the three methods of"tonifying the spleen and stomach,removing stasis poison and clearing cancer poison"are established,and the flexible fusion application with the syndrome reflects the concept of"always strengthening the body resistance,not forgetting to attack evil and combining prevention and treatment".Two cases of gastric cancer with emaciation after surgery were treated by tonifying the spleen to produce muscle and eliminate distention,removing blood stasis and detoxifying cancer,but each had its own emphasis on strengthening the body resistance and eliminating evil.After treatment,all the cases were stable,and no recurrence and metastasis were observed during follow-up period.[Conclusion]Professor CHEN treats gastric cancer by"tonifying the spleen and stomach,removing stasis poison and clearing cancer poison",which has good curative effect and certain clinical practical value.

[Objective]To summarize the clinical experience of Professor LU Zheng,in the treatment of colon cancer by"strengthening spleen and clearing intestine and detoxifying".[Methods]Through outpatient follow-up study,case collection and collation,looking up the literature and TCM classics related to colon cancer,studying Professor LU Zheng's"DU Zheng Opinions",combining with clinical interpretation,this article discussed the etiology and pathogenesis of colon cancer,explained the treatment method of colon cancer as"strengthening spleen and clearing intestines and detoxifying"under the guidance of"four-layer general principle of toxin syndrome",summarized the key points of clinical syndrome differentiation and treatment and the rules of prescription and drug use,finally provided examples to illustrate it.[Results]Professor LU Zheng believed that the basic pathogenesis of colon cancer was the damage of spleen and stomach vitality and the accumulation of cancer toxin in the intestine,and further pointed out that the damage of spleen and stomach vitality was the premise of the occurrence and development of colon cancer,and the accumulation of cancer toxin in the intestine was the key to the occurrence,recurrence and metastasis of colon cancer.The clinical treatment should focus on restoring the vitality of spleen and stomach,clearing the endogenous evil factors such as dampness,heat,phlegm and stasis in the intestine and the residual cancer toxin,clearing the intestinal Qi mechanism,that was,taking"strengthening spleen and clearing intestines and detoxifying"as the guiding ideology,according to the specific symptoms of patients after operation,it was summarized into five syndromes:damp-heat toxin,phlegm and stasis toxin,stasis and blood toxin,Yang deficiency cold toxin,Yin deficiency heat toxin.According to the concept of supporting the essence and removing the toxin,the treatments were heat-clearing and detoxification,phlegm-removing and stasis-removing,stasis-removing and detoxification,warming the essence and removing the toxin,nourishing the Yin and detoxifying,thus to eliminate the postoperative residual toxin,completely eradicate new cancer toxin.After the treatment of"strengthening spleen,clearing intestine and detoxifying toxin",the symptoms were relived and quality of life was improved,and there was no tumor recurrence and metastasis during follow-up period.[Conclusion]By summarizing the clinical experience of Professor LU Zheng's"strengthening spleen and clearing intestine and detoxifying"in the treatment of colon cancer,the application of LU Zheng essences theory of toxifying syndrome in the treatment of cancer was demonstrated,and the reference was provided for the differentiation and treatment of colon cancer.Professor LU Zheng takes"strengthening spleen and clearing intestine and detoxifying"as the basic rule to treat colon cancer,which can provide a reference for the prevention and treatment of colon cancer with traditional Chinese medicine.

Objective To investigate the effect of type 2 dengue virus(DENV-2)infection on autophagy in human umbilical vein endothelial cells(HUVECs)and the mechanism mediating the inhibitory effect of baicalin against DENV-2 infection.Methods Cultured HUVECs with DENV-2 infection were treated with different concentrations of baicalin,and the changes in autophagy of the cells were detected using transmission electron microscopy.Lyso Tracker Red staining was used to examine pH changes in the lysosomes of the cells,and the expressions of ATG5,beclin-1,LC3,P62,STX17,SNAP29,VAMP8,and PI3K/AKT signaling pathway-related proteins were detected by Western blotting.DENV-2 replication in the cells were evaluated using RT-qPCR.The differentially expressed proteins in DENV-2-infected HUVECs were identified by proteomics screening.Results Treatment with baicalin did not significantly affect the viability of cultured HUVECs.Proteomic studies suggested that the PI3K-AKT pathway played an important role in mediating cell injury induced by DENV-2 infection.The results of RT-qPCR demonstrated that baicalin dose-dependently inhibited DENV-2 replication in HUVECs and produced the strongest inhibitory effect at the concentration of 50 μg/mL.Transmission electron microscopy,Lyso Tracker Red staining,RT-qPCR,and Western blotting all showed significant inhibitory effect of baicalin on DENV-2-induced autophagy in HUVECs.DENV-2 infection of HUVECs caused increased cellular expressions of LC3 and P62 proteins,which were significantly lowered by treatment with LY294002(a PI3K inhibitor).Conclusion Baicalin inhibits DENV-2 replication in HUVECs and suppresses DENV-2-induced cell autophagy by inhibiting the PI3K/AKT signaling pathway.

Reperfusion injury occurs after return of spontaneous circulation (ROSC) in patients with cardiac arrest (CA), which leads to multiple organ dysfunction, called post-cardiac arrest syndrome (PCAS). PCAS is closely related to the prognosis of CA patients, and is an independent risk factor of survival. Integrated traditional Chinese and Western medicine diagnosis and treatment is critical for improving prognosis of PCAS. In order to guide and standardize integrated traditional Chinese and Western medicine diagnosis and treatment in PCAS among clinicians, nurses and research personnel in China, the Emergency Medicine Professional Committee of the Chinese Society of Integrated Chinese and Western Medicine has established an expert group to determine 14 clinical issues related to the diagnosis and treatment of PCAS with integrated traditional Chinese and Western medicine through clinical survey. The working group formulates a search strategy for each clinical issue according to the PICO principle. Chinese and English literature were searched from CNKI, Wanfang, VIP, SinoMed, PubMed, Embase, and Cochrane Library. The grade of recommendations assessment, development and evaluation (GRADE) were used to form the level of evidence and recommendation. When the literature evidence was insufficient, the recommendations and level of recommendation were formed after expert discussion. Combined with the aspects of generalizability, suitability, and resource utilization, the expert consensus developed 28 recommendations around the 14 aspects of three stages of PCAS, including early circulation, respiratory support and reversible cause relief, mid-term neuroprotection, improvement of coagulation, prevention and treatment of infection, kidney and gastrointestinal protection and blood sugar control, post rehabilitation treatment, providing references for the integrated traditional Chinese and Western medicine of the diagnosis and treatment for PCAS.

Dysfunction of the Hippo pathway enables cells to evade contact inhibition and provides advantages for cancerous overgrowth. However, for a significant portion of human cancer, how Hippo signaling is perturbed remains unknown. To answer this question, we performed a genome-wide screening for genes that affect the Hippo pathway in Drosophila and cross-referenced the hit genes with human cancer genome. In our screen, Prosap was identified as a novel regulator of the Hippo pathway that potently affects tissue growth. Interestingly, a mammalian homolog of Prosap, SHANK2, is the most frequently amplified gene on 11q13, a major tumor amplicon in human cancer. Gene amplification profile in this 11q13 amplicon clearly indicates selective pressure for SHANK2 amplification. More importantly, across the human cancer genome, SHANK2 is the most frequently amplified gene that is not located within the Myc amplicon. Further studies in multiple human cell lines confirmed that SHANK2 overexpression causes deregulation of Hippo signaling through competitive binding for a LATS1 activator, and as a potential oncogene, SHANK2 promotes cellular transformation and tumor formation in vivo. In cancer cell lines with deregulated Hippo pathway, depletion of SHANK2 restores Hippo signaling and ceases cellular proliferation. Taken together, these results suggest that SHANK2 is an evolutionarily conserved Hippo pathway regulator, commonly amplified in human cancer and potently promotes cancer. Our study for the first time illustrated oncogenic function of SHANK2, one of the most frequently amplified gene in human cancer. Furthermore, given that in normal adult tissues, SHANK2's expression is largely restricted to the nervous system, SHANK2 may represent an interesting target for anticancer therapy.

Objective To investigate the clinical characteristics in nonalcoholic steatohepatitis (NASH),to provide the basis for the early clinical diagnosis.Methods 200 cases of non-alcoholic fatty liver disease(NAFLD) were divided into NAFL group(115 cases) and NASH group(85 cases).Age,gender,body mass index,blood pressure,clinical symptoms,accompany illnesses,biochemical and image index in the two groups were analyzed and compared retrospectively.Results Asthenia,accounted for 40％,which was the most common gastrointestinal symptom of NASH group.and abdominal distension,anorexia,nausea and vomiting,liver area pain and liver were all involved.There were no significant differences in mainly gastrointestinal symptoms between patients with NASH and NAFL group(P ＞ 0.05),but no symptoms incidence of the two group were higher (44.7％ vs 49.2％ ％ P ＞ 0.05) ;the incidence of obesity,hyperlipidemia,type 2 diabetes,hypertension in NASH group were significantly different compared with NAFL(45.9％ vs 20％,41.2％ vs 22.3％,28.2％ vs 15.6％ respectively P ＜ 0.05) ; In NASH group,BMI,fasting glucose(FPG),2hPPG,serum ferritin,hyaluronic acid,Ⅳ collageninsulin resistance index (HOWA-IR)increased significantly compared with NAFL[(28.68 ± 0.92)vs (22.21 ± 0.43),(9.63 ± 0.64)mmol/L vs (4.92 ± 0.78)mmol/L,(12.96 ±0.28) mmol/L vs (7.04 ±0.13) mmol/L,(243.56 ±7.95) ng/mL vs (140.03 ± 6.80)ng/mL,(130.26 ±9.i6)ng/mL vs (74.85 ±6.54)ng/mL,(130.56 ±8.16)ng/mL vs (72.68 ±7.24) ng/mL,(5.36±0.45) vs (2.63 ±0.12),respectively P＜0.05)].Conclusion Patients with NASH had no obvious gastroenterology symptoms.Obesity,type 2 diabetes,hypertension are more with NASH,and there may be multiple metabolic disorders.

The instrument is com posed of such components as SCM,pressure sensor,electric pump,magnetic valve and sleeve-type gasbag.It can control the hemostatic pressure,hemostatic time and slack time automatically.So the ischemic necrosis can be avoided thoroughly thr ough it.With the small volume,light weight,low power consumption and convenient operation,the instrument is fit for emergency treatment of mass casualties.

Objective To study the pathogenesis of osteoporosis in patients with liver cirrhosis following hepatitis B virus(HBV) infection(HBV-liver cirrhosis).Methods Bone mineral density(BMD) was measured by NM-300 sigle-energy X-ray acsorptiometry system for 61 patients with HBV-liver cirrhosis and 30 age-matched healthy controls.Serum levels of calcium regulatory hormones(CRH),1,25-dihydroxycholecalciferol [1,25(OH)_2D_3],parathyroid hormone(PTH),calcitonin(CT),osteocalcin(BGP),interleukin-1?(IL-1?),interleukin-6(IL-6) and tumor-necrosis factor-?(TNF-?),and urine level of crosslaps were measured in these patients and controls.Patients with HBV-liver cirrhosis were divided into two groups,osteoporotic and non-osteoporotic,based on their BMD in the ulna and radius,for comparison.Results BMD of the ulna and radius in patients with HBV-liver cirrhosis were significantly lower than those in controls.Serum levels of 1,25(OH)_2D_3 and BGP in patients with HBV-liver cirrhosis were significantly lower than those in controls,and much lower in osteoporotic group than that in non-osteoporotic one.While urine level of crosslaps increased significantly in patients with HBV-liver cirrhosis than in controls, much more increased in the those with osteoporosis than those without osteoporosis.Serum level of 1,25(OH)_2D_3 correlated with serum levels of BGP and BMD of the ulna and radius.Urine level of crosslaps correlated inversely with BMD of the ulan and radius in osteoporotic patients,but not in non-osteoporotic ones.Serum levels of IL-1?,IL-6 and(TNF-?) and urine level of crosslaps increased significantly in patients with HBV-liver cirrhosis than in controls,as well as increased significantly in those with osteoporosis than in those without osteoporosis.Serum levels of IL-1?,IL-6 and TNF-? and urine level of crosslaps correlated inversely with BMD of the ulna,particularly more significantly in those with osteoporosis than those without osteoporosis.Conclusions Two factors,less osteogenesis and more osteolysis,existed in patients with HBV-liver cirrhosis.Serum level of 1,25(OH)_2D_3 and serum levels of IL-1?,IL-6 and(TNF-?) played an important role in weakened bone formation and in increased bone resorption,respectively.Adequate supplementation of vitamin D_3 and reducing serum levels of IL-1?,IL-6 and TNF-? may be very important for prevention and treatment of hepatic bone disease.

Objective To investigate the diagnostic values of color ultrasound to the stenosis of extracranial carotid arteries.Methods Color ultrasound examination and digital subtraction angiography(DSA)were performed in 145 patients with ischemic cerebravascular disease.The sensitivity and specificity of color ultrasound for the stenosis of extracranial carotid arteries were analyzed according to the golden standard of DSA results.Results The sensitivity and specificity of color ultrasound for stenosis of carotid bifurcation,subclavicular artery and vertebral artery were 96.8%,80.6%,60.0% and 98.7%,33.3%,89.0%.The accuracy of color ultrasound for stenosis of carotid bifurcation,subclavicular artery and vertebral artery were 93.0%,92.8% and 82.1%.Conclusion Color ultrasound examination are effective for the diagnosis of carotid bifurcation sterosis,and less effective for the diagnosis of the subclavicular and vertebral artery stenosis because of the poor sensitivity.