Objective To investigate the effect of silent information regulator 1 (SIRT1)on cigarette smoke extract (CSE)-induced apoptosis of human alveolar epithelial cell (AECs).Methods The expression levels of SIRT1 protein were examined by Western Blotting in A549 cells that were treated with CSE at different concentrations.The impairment models of A549 cells induced by CSE were established.The concentration of CSE was 20.0％ and the treatment time of CSE was 24 hours.A549 cells were pretreated with 20 μmol/L resveratrol(Res)or 5 mmol/L nicotinamide (NAM) for 2 h before CSE treatment.The protein levels of SIRT1,Bax and Bcl2 were further explored by Western blotting.The proportion of apoptotic A549 cells was measured using MTT.Results The expression of SIRT1 was reduced after treatment with CSE in a dose-dependent manner(P＜0.05).Compared with the control group,the group of CSE treatment with 2.5％,5.0％,10.0％,20.0％and 40.0％ CSE for 24h showed that the expression of SIRT1 in A549 cells was decreased by 25.6％(P＜0.05),35.2％(P＜0.05),38.7％ (P＜0.05),57.9％ (P＜0.05)and 64.0％(all P＜0.05)separately; that A549 cell viability was decreased in a dose-dependent manner; and that A549 cell viability was decreased by 10.2％(t=2.035,P＜0.05),18.4％(t=4.269,P＜0.05),27.7％ (t=5.963,P＜0.05),59.0％(t=21.140,P＜0.05)and 88.1％(t=58.827,P＜0.05)separately.CSE plus 20 μmol/L Res pretreatment reversed the expression levels of SIRT1,Bax and Bcl2 in A549 cells (all P＜0.05)and reduced the apoptosis of A549 cells.The effects of CSE on inhibiting SIRT1 pathways were aggravated by NAM (an inhibitor of SIRT1) in the A549 cells (P＜ 0.05).Conclusions SIRT1 plays important role in regulating the apoptosis of human alveolar epithelial A549cell induced by CSE.SIRT1 may inhibit apoptosis by up-regulating Bcl2 expression and downregulating Bax expression,which has a protective effect on A549 cells apoptosis induced by CSE.

Objective To study the rule of distribution and evol vement of TCM syn dromes in HIV/AIDS patients. Methods Totally 177 HIV/AI DS patients were investigated with questionnaire in the epide miological study to collect the data of diagnosis, and the rou tine test of CD4+T lymphocyte count was carried out. Results The young and mid dle-aged females were the most involved group with an increasing incidence in G u angdong Province and with a strong relation of sex and infection route. The main symptoms of the 177 HIV/AIDS patients were fatigue, cough and loss of appetite, the secondary ones were night sweating, skin itching, headache, and hair loss, and the most common sign was skin rash. The leading TCM syndromes were qi and yi n deficiency and lung and kidney deficiency. The distribution of symptoms and si gns, syndromes and various indices was significantly different in gender, age an d infection route (P

Objective To detect the expression of tumor‐associated macrophages(TAM) in esophageal squamous cell carci‐noma ,and to study the clinical pathological characteristics of T AM in the esophageal squamous cell carcinoma .Methods Patients with esophageal squamous cell carcinoma who accepted operation were chosen as study subjects ,and tissue samples of esophagus were collected ,including 90 squamous cell carcinoma tissues ,20 paracancerous atypical hyperplasia and 20 normal mucosa tissues , and the expression of CD206 ,MCP‐1 were detected by immunohistochemisty .Results The positive expression of CD206 was sig‐nificantly increased in tissues of esophageal squamous cell carcinoma (P<0 .01) ,and it was positively correlated with clinical stage , invasion depth and lymph node metastasis in esophageal squamous cell carcinoma (P<0 .05) .The expression of MCP‐1 was signifi‐cantly increased in esophageal squamous carcinoma tissues (P<0 .05) ,and its positive expression was closely correlated with depth of invasion and lymph node metastasis in esophageal squamous cell carcinoma (P< 0 .05) .There was a positive relation between ATM infiltration quantity and the expression of MCP‐1(r=0 .617 ,P<0 .05) .Conclusion The positive expression of TAM was up‐regulated in esophageal squamous cell carcinoma tissues ,and its number was positively correlated with clinical stage ,invasion depth and lymph node metastasis .

Objective To evaluate the gene amplification and protein expression of HER-2 in gastric carcinoma with or without neuroendocrine differentiation (NED), and to explore the difference in HER-2 gene profile between these two neo?plasms. Methods Cases of gastric carcinoma with NED (n=70) and cases of gastric carcinoma without NED (n=150) were retrospectively reviewed. Gene amplification and protein expression of HER-2 genes in gastric carcinoma with or without NED were detected by combination of immunohistochemical method (IHC) and double color silver dye for in situ hybridiza?tion (DISH). Prognosis of gastric cancer patients with NED were predicted using Kaplan-Meiers survival analysis. Results Expression rates of HER-2 in gastric cancer with and without NED are 20.00%and 21.33%respectively. Amplification of HER-2 in gastric cancer with and without NED are 8.57%and 14.67%respectively. Gene amplification and protein expres?sion of HER-2 between gastric cancer with NED and without NED showed no statistical difference. Chromosome 17 multi-body positively correlated with gene HER-2 amplification in Gastric carcinoma with NED. Postoperative survival period in patients of gastric carcinoma with NED and HER-2 amplification was shorter than that in patients of gastric carcinoma with?out NED but with HER-2 amplification. Gastric carcinoma with or without NED, HER-2 gene amplification, lymph node me?tastasis and operation method obviously correlate with prognosis of gastric carcinoma patients (P<0.05). Conclusion The gastric cancer with NED is a special type of gastric cancer, there was no difference of gene amplification and protein expres?sion of HER-2 gene between gastric carcinoma with NED and without NED. Poor prognosis would be expected in gastric can?cer patients with NED and HER-2 amplification.

Objective To explore the seed protein extraction method for Amomum villosum Lour., so as to lay the foundation for the study of differential proteomics of Amomum villosum Lour. during seed dormancy and germination. Methods We used the four kinds of commonly-used protein extraction methods for the experiment, including Tris-HCl extraction method, trichloroacetic acid (TCA) /acetone precipitation method, direct lysis buffer method and improved Tris-saturated phenol method. The obtained proteins were quantified, and then were analyzed by sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) . Results The protein yield was the highest when using direct lysis buffer method, which was mainly composed of small molecule protein. The protein yield was the lowest using Tris/HCl method, characterized by the diffused stripes in SDS-PAGE spectra. The protein yield was moderate using TCA/acetone method and improved Tris-saturated phenol method, and the obtained protein by the TCA/acetone method had less stripes in SDS-PAGE spectra than that by Tris-saturated phenol method, and was characterized by macromolecular protein. Conclusion The combination of direct lysis buffer extraction method and improved Tris-saturated phenol method may provide reference for seed protein extraction method of two-dimensional electrophoresis for Amomum villosum Lour..

Objective:To investigate the effect of miR-218 on the proliferation and invasion of breast cancer cells.Methods: The expression of miR-218 in breast cancer tissues and breast cancer cell lines was detected by qPCR.The relationship between the expression of miR-218 and the clinicopathological parameters of breast cancer were analyzed.Double luciferase assay was used to detect the relationship between miR-218 and SNX4.MTT assay and invasion assay were used to detect the proliferation and invasion of breast cancer cells after overexpression of miR-218.MTX assay and invasion assay were used to detect the recovery level of SNX4 on the proliferation and invasion of breast cancer cells.The effect of miR-218 on the tumorigenesis of breast cancer cell lines was examined by tumorigenesis in nude mice.Results: The expression level of miR-218 in breast cancer tissue and MCF-7 cell line was higher.The expression of miR-218 was associated with pathological stage of breast cancer and lymph node metastasis.SNX4 may be the target of miR-218.Overexpression of miR-21 could inhibit the proliferation and invasion of breast cancer cells.Overexpression of SNX4 could reverse the inhibitory effect of miR-218 on breast cancer cells.Overexpression of miR-218 could inhibit the breast cancer cell line in nude mice tumorigenic ability.Conclusion: miR-218 can up-regulate the expression of miR-218 in breast cancer,and miR-218 can regulate the expression of SNX4 in breast cancer cell proliferation and invasion.

Circular RNAs (circRNAs) are a type of endogenous non-coding RNAs discovered recently, which are produced by models such as circularization driven by intron pairing and are regulated by related regulatory factors. An important biological function of circRNAs is to act as a molecular sponge to make the level of microRNAs significantly change in a short period of time. A number of studies have shown that circRNAs are closely associated with ischemic stroke. Therefore, a better understanding of the function of circRNAs and its regulatory mechanism will help to explore the expression of circRNAs in early ischemic stroke and its molecular mechanism in the occurrence and development of ischemic stroke, so as to establish an early warning model to realize the early detection, early diagnosis and early treatment of ischemic stroke. This article reviews the definition, biological characteristics, and expression changes and roles of circRNAs after ischemic stroke.

Objective To evaluate the effect of isoflurane preconditioning on inflammatory responses during spinal cord injury (SCI ) in rats .Methods Sixty adult male Sprague-Dawley rats ,weighing 250-300 g , were randomly divided into 3 groups ( n= 20 each ) using a random number table :sham operation group (S group) , SCI group , and isoflurane preconditioning group (I group ) . The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg .SCI was produced by a weight-drop contusion at the T10 level .The rats inhaled 2% isoflurane for 2 h ,and the model was established at 24 h after the end of isoflurane inhalation in I group . Neurological function was assessed and scored by using the the Basso , Beattie , Bresnahan (BBB ) Locomotor Rating Scale on 7 days after SCI .Five rats in each group were then chosen and spinal cord specimens were obtained and cut into sections which were stained with haematoxylin and eosin for determination of the viable neuron count .Fifteen rats in each group were sacrificed and the spinal cord was removed for detection of nuclear factor kappaB (NF-κB ) and interleukin-1β (IL-1β) expression (by Western blot ) .Results Compared with S group ,BBB score and the number of viable neurons were significantly decreased ,and the expression of NF-κB and IL-1βprotein was up-regulated in SCI group ( P<0.05) .Compared with SCI group ,BBB score and the number of viable neurons were significantly increased ,and the expression of NF-κB and IL-1βprotein was down-regulated in group I ( P<0.05 ) .Conclusion The mechanism by which isoflurane preconditioning protects the spinal cord is related to inhibition of inflammatory responses in rats .

Background The pathogenesis of diabetic retinopathy (DR) might be related with oxidative stress and its mechanism has not been fully elucidated.Grape seed proanthocyanidin extracts (GSPE),a powerful antioxidant,plays roles in some systemic diseases.However,the effect and mechanism of GSPE in DR has not been illuminated clearly.Objective This study was to investigate whether GSPE has a protective effect on the retinas of diabetic subjects and explore its mechanism.Methods Thirty SPF adult Wistar rats were divided into the control group,diabetic group and diabetes + GSPE group according to the randomized number table.Diabetic models were induced by intraperitoneal injection of 100 mg/kg strebtozotocin (STZ) prepared with citrate buffer,and only the equal amount of citrate buffer was used in the same way in the control group.GSPE solution was intragastrally used 250 mg/kg daily in the rats of the diabetes+GSPE group from injective day through 8 weeks,and distilled water was used in the same way in the rats of the control and diabetic groups.The rats were sacrificed in the eighth week after injection and the retinas were isolated.The morphology of the retina were examined by hematoxylin and eosin stain.Retinal homogenatewas prepared for the assay of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) activity and malonaldehyde (MDA) content.The expressions and location of nuclear erythroid related factor 2 (Nrf2) was determined by immunochemistry,and the expressions of Nrf2 and heme oxygenase-1 (HO-1) were quantitatively analyzed by Western blot.The apoptosis of retinal cells was detected by TUNEL.Results In the eighth week after modeling,the blood glucose levels were significantly higher and the body weight was lower in the rats of the diabetic group and diabetes+GSPE group compared with the control group (all at P＜0.01).Retinal structure was normal in the rats of the control group.However,loose tissue,irregular arrangement of cells and thickness decrease of retinal fiber layer,retinal ganglion cell layer and inner plexiform layer were exhibited in the rats of the diabetic group,while the morphology abnormality was slight in the rats of the diabetes+GSPE group.The SOD and GSH-Px activities and MDA content were significantly different among the 3 groups (F =11.010,P =0.001 ; F =12.072,P =0.000 ; F =25.224,P=0.000),and activities of SOD and GSH-Px in the retinas were significantly lower,and the MDA level was higher in the rats of diabetic group than those of the diabetes+GSPE group and the control group (all at P＜0.01).The relative expressing levels of Nrf2 were (165.5±29.4) ％ and (134.8 ±7.8) ％ in the diabetes+ GSPE group and diabetic group,with a significant difference between them (t=2.450,P=0.044).Compared with the diabetic group,the expressing level of the HO-1 was sigficantly increased ([170.2±22.0)％ versus [125.3±9.2] ％,t =2.360,P =0.002).TUNEL showed that the retinal apoptotic cells of diabetic rats were mainly located in the retinal fiber layer,RGCs layer,inner and outer nuclear layer,and the number of apoptotic cells was less in the diabetes+GSPE group compared with the diabetic group under the fluorescence microscope.Conclusions GSPE can play a protective role on diabetic rats by activating Nrf2 pathway and therefore improving retinal oxidative stress and decreasing apoptosis.

Objective Studying the status quo and constraints for rural healthcare service in grassroot rural healthcare units,for policy recommendations.Methods Using data from the fourth healthcare service investigation,by means of quantitative interview and qualitative interview,for an investigative interview of 348 township hospitals and 251 village clinics in 31 provinces in China.Results Deployment percentage of primary heahhcare services at township hospitals level is 49.1% (28.0/57),and that for village clinics is 60.6%(5.4/9).Conclusions The key to upgrading rural healthcare service system at grassroots level is to deploy better diagnostic equipments,upgrade the diagnostic competence of grassroots healthcare personnel and build a continuous service system for primary healthcare service.