Objective To examine the effect and mechanism of Duhuo Jisheng Decoction on lumbar intervertebral disc degeneration.Methods In total,105 Sprague-Dawley(SD)rats were randomly assigned to seven groups:sham operation group,model group,Duhuo Jisheng Decoction 1,rapamycin group,rapamycin+Duhuo Jisheng Decoction group,MHY1485 group,and MHY1485+Duhuo Jisheng Decoction group.Except for the sham operation group,the other groups underwent annulus fibrosus puncture to establish a lumbar intervertebral disc degeneration(IDD)animal model.After successful model establishment,a six-week drug intervention was performed,followed by MRI evaluation of the degree of disc degeneration.Samples of the L5-6 intervertebral disc were collected for HE and Alcian blue-nuclear fast red staining,and the degeneration of the nucleus pulposus and changes in the extracellular matrix were observed using light microscopy.Simultaneously,the expression levels of the downstream proteins of the mTOR pathway,p70S6K and 4E-BP1,along with the autophagy-related genes Beclin-1 and LC3,were assessed at both the protein and mRNA levels.Autolysosomes in nucleus pulposus cells were visualized using transmission electron microscopy(TEM).Results ①MRI showed disc Pfirrmann grade I in the sham group,and disc Pfirrmann grade Ⅲ-Ⅳ in the remaining 6 groups in L5-6 segments.②The degree of lumbar disc degeneration with histomorphological observation:MHY1485 group>model group>MHY1485 group+Duhuo Jisheng Decoction group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group.Allan-nuclear red staining extracellular matrix proteoglycan content:sham group>rapamycin+Duhuo Jisheng Decoction group>rapamycin group>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485 group.③Relative expression of p-mTOR,p70S6K and 4E-BP1 downstream of mTOR signaling pathway:MHY1485 group>MHY1485 group+Duhuo Jisheng Decoction group>model group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group,each experimental group varied significantly from the model group(P<0.01).④Autophagy-related protein Beclin-1 and LC3 expression levels:rapamycin+Duhuo Jisheng Decoction group>Duhuo Jisheng Decoction group>rapamycin>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485>sham group,and the content of each group was significantly(P<0.01).⑤TEM observation of autophagy levels in the cells of each group showed the formation of autolysosomes in Duhuo Jisheng Decoction group,rapamycin group andrapamycin+Duhuo Jisheng Decoction group.Conclusion Duhuo Jisheng Decoction can slow down the degenerative process of lumbar intervertebral discs in rats,and its effect may be linked to the suppression of the mTOR signaling pathway and the promotion of autophagy in nucleus pulposus cells.

Objective To examine the effect and mechanism of Duhuo Jisheng Decoction on lumbar intervertebral disc degeneration.Methods In total,105 Sprague-Dawley(SD)rats were randomly assigned to seven groups:sham operation group,model group,Duhuo Jisheng Decoction 1,rapamycin group,rapamycin+Duhuo Jisheng Decoction group,MHY1485 group,and MHY1485+Duhuo Jisheng Decoction group.Except for the sham operation group,the other groups underwent annulus fibrosus puncture to establish a lumbar intervertebral disc degeneration(IDD)animal model.After successful model establishment,a six-week drug intervention was performed,followed by MRI evaluation of the degree of disc degeneration.Samples of the L5-6 intervertebral disc were collected for HE and Alcian blue-nuclear fast red staining,and the degeneration of the nucleus pulposus and changes in the extracellular matrix were observed using light microscopy.Simultaneously,the expression levels of the downstream proteins of the mTOR pathway,p70S6K and 4E-BP1,along with the autophagy-related genes Beclin-1 and LC3,were assessed at both the protein and mRNA levels.Autolysosomes in nucleus pulposus cells were visualized using transmission electron microscopy(TEM).Results ①MRI showed disc Pfirrmann grade I in the sham group,and disc Pfirrmann grade Ⅲ-Ⅳ in the remaining 6 groups in L5-6 segments.②The degree of lumbar disc degeneration with histomorphological observation:MHY1485 group>model group>MHY1485 group+Duhuo Jisheng Decoction group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group.Allan-nuclear red staining extracellular matrix proteoglycan content:sham group>rapamycin+Duhuo Jisheng Decoction group>rapamycin group>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485 group.③Relative expression of p-mTOR,p70S6K and 4E-BP1 downstream of mTOR signaling pathway:MHY1485 group>MHY1485 group+Duhuo Jisheng Decoction group>model group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group,each experimental group varied significantly from the model group(P<0.01).④Autophagy-related protein Beclin-1 and LC3 expression levels:rapamycin+Duhuo Jisheng Decoction group>Duhuo Jisheng Decoction group>rapamycin>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485>sham group,and the content of each group was significantly(P<0.01).⑤TEM observation of autophagy levels in the cells of each group showed the formation of autolysosomes in Duhuo Jisheng Decoction group,rapamycin group andrapamycin+Duhuo Jisheng Decoction group.Conclusion Duhuo Jisheng Decoction can slow down the degenerative process of lumbar intervertebral discs in rats,and its effect may be linked to the suppression of the mTOR signaling pathway and the promotion of autophagy in nucleus pulposus cells.

Macrophages are important cells of the immune system. Tumor-associated macrophages are enriched macrophages near tumor cells or tissues. Their role is mainly to promote the construction of tumor inflammatory microenvironment and inhibit tumor immune response. Cell co-culture system is a symbiotic culture system formed by mimicking the internal environment of the body in vitro. The co-culture condition is relatively consistent with the environment in vivo, enabling better information exchange and material exchange between cells, which is a supplement to the monolayer cell culture and animal experiments. Tumor-associated macrophages and tumor cells co-exist in the tumor microenvironment. Thus, constructing a co-culture system for tumor-associated macrophages and tumor cells would be conducive to studying the antitumor effect of tumor-associated macrophages and developing new immunotherapy drugs. The co-culture system would provide a new direction for treating malignant tumors. This article mainly reviewed the co-culture patterns of macrophages and the antitumor effects of different phenotypes of macrophages, and highlighted the importance of using immunotherapy to treat malignant tumors in the tumor microenvironment.

Objective: To investigate the effects of sulforaphane (SFN) on proliferation , migration and invasion of human renal carcinoma cells and its mechanism. Methods: The cultured human renal carcinoma cells 786⁃O were divided into control group (0 μmol/L) and SFN group (5 , 10 , 20 μmol/L) . The activated proliferation of cells was detected by CCK⁃8 ; the effect of SFN on migration of 786⁃O cells was detected by scratch healing assay and Transwell cell migration assay; the effect of SFN on the invasion ability of 786⁃O cells was detected by Transwell cell invasion ability assay; Western blot and qRT⁃PCR were used to detect the effects of SFN on the expression of epithelial⁃mesenchymal transition (EMT) Ⅳrelated proteins and mRNA. The effect of SFN on the expression of NF⁃κB signaling pathway was detected by Western blot. Results: After SFN treatment for 24 , 48 and 72 h , the proliferation activity of 786⁃O cells decreased with the increase of SFN concentration ; compared with the control group , the cell migration and invasion ability of SFN⁃treated group were significantly reduced ; with the increase of SFN concentration , the mRNA and protein expression levels of E ⁃cadherin in 786⁃O cells increased , while the mRNA and protein expression levels of N ⁃cadherin and Vimentin decreased ; the levels of NF⁃κB signaling pathway related protein phosphorylated p65 and phosphorylated IκBα decreased with the increase of SFN concentration. Conclusion SFN may inhibit the proliferation , migration and invasion of human renal carcinoma cells by regulating the EMT process of renal carcinoma through inhibition of NF⁃κB signaling pathway.

Objective To investigate the clinical application value of pulp mummification therapy and root canal therapy in senile chronic pulpitis. Methods The clinical data of elderly patients with chronic pulpitis who were admitted to our department from January 2016 to January 2017 were analyzed retrospectively. According to the different treatment methods, 120 elderly patients were divided into two groups: the pulp mummification therapy group and the root canal group. The clinical efficacy of the two groups were observed and the patients were followed up for a period of six months. The prognosis of the patients was recorded and counted. Results There were 60 patients in the pulp mummification therapy group, of which 34 were markedly effective, 19 were effective and 7 were ineffective. There were 60 patients in the root canal treatment group, of which 33 were markedly effective, 17 effective and 10 ineffective. There was no significant difference between data of the two groups (P＞0. 05). Six months after treatment, the patients were followed up for half a year. There were 2 adverse reaction cases in 60 patients of the pulp mummification therapy group. The incidence of adverse reactions was 3. 33％. Among the 60 patients in the root canal treatment group, 1 patient developed adverse reactions, The incidence of adverse reactions was 1. 67％. There was no difference in the incidence of adverse reactions between the two groups (P＞0. 05). Conclusion Both pulp mummification therapy and root canal therapy have a good clinical effect on elderly patients with chronic pulpitis. However, the pulp mummification therapy is suitable for patients whose radicular pulp have not been infected and the root canal is suitable for patients whose radicular pulp have been infected. Therefore, appropriate treatment should be selected based on patient characteristics to improve the treatment effect during the process of treatment.

Objective To assess the impact of miR-429 on lung adenocarcinoma cell SPC-A1 growth inhibition.Methods Pre-miRTM miR-429 precursor was synthesized and transfected to the SPC-A1 cells by liposome; qRT-PCR assay was used to quantify the miR-429 expression levels; The proliferation of SPC-A1 cells was evaluated by Cell Counting Kit-8 (CCK8).The cell apoptosis was evaluated by Annexin V Assay; The cell cycles of each group were assayed by flow cytometry;Western-blot was used to analyze the expression of cylines.Results The expression level of miR-429 was highly induced after transfection (P ＜ 0.001) ;CCK-8 assay showed the cell proliferation activity of pre-miR-429 group was lower than that of blank and control group 48h and 72 h after transfection(P =0.0167,0.0383,P =0.0320,0.0465),whereas the apoptosis rate had no significant difference between pre-miR-429 and control 24h after transfection by Annexin V Assay(P ＞ 0.05) ; The flow cytometry at 48h after transfection showed that miR-429 decreased the percentage of cells in G1 phase,but increased in S phase,indicating the cell cycle arrest at S phase(P =0.0010,0.0010 ; P =0.0068,0.0133) ; however,the expression level of Cyclin E in pre-miR-429 group had no difference compared with control.Conclusion miR-429 could inhibit cell proliferation and promote cell cycle arrest of lung adenocarcinoma cell SPC-A1.miR-429 may play a potential tumor suppressor role in lung adenocarcinoma cell SPC-A1.

Objective To study the microbial strains,risk factors and resistance profiles of lower respiratory tract fungal infection in hospital of Zhoushan archipelago area.Methods A total of 204 patients who were hospitalized for lower respiratory tract infection were retrospectively analyzed from May 2008 to April 2011 in Zhoushan archipelago area,and collected 204 fungal strains isolated from confirmed lower respiratory tract fungal infection cases.Chi-square test and Logistic regression analysis were performed.Results Among the 204 fungal strains isolated from lower respiratory tract specimens,110 (53.8％) strains of Candidaalbicans,32 (15.7％) strains of Candida tropicalis,24 (11.8％) strains of Candida glabrata,12 (5.9％) strains of Candida krusei,14 (6.9％) strains of other Candida,and 12 (5.9％) strains of Aspergillus were detected.Logistic regression analysis showed that chronic obstructive pulmonary disease,bacterial pneumonia,long-term use of broadspectrum antibiotics and corticosteroids,endotracheal intubation or incision,old age,exposure in intensive care unit (ICU),and hospitalization ≥7 days were major risk factors (P=0.000,0.001,0.000,0.000,0.012,0.000,0.000,0.000).The resistance rates of isolated Candida against amphotericin B,5-flucytosine,voriconazole,itraconazole and fluconazole were 0,2.1％,4.2％,14.8％ and 22.9％,respectively.Conclusions Candida albicans is the major pathogen of lower respiratory tract fungal infection in hospital of Zhoushan archipelago area,and Candida is sensitive to amphotericin B,5-flucytosine and voriconazole.

Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.

Objective To investigate iodine nutrition and thyroid health status among residents in Zhoushan archipelago, and to analyse their relationship.Methods A total of 3 284 residents in Zhoushan archipelago were surveyed by questionnaire and their thyroids were examined by B-mode ultrasound.The levels of urinary iodine and thyroid function were detected.Results The median level of urinary iodine in 3 284 residents was 226.0 μg/L, being 320.7 μg/L in citizens, 188.9 μg/L in farmers, 122.2 μg/L in salt-makers, 193.6 μg/L in fishers, and 271.7 μg/L in buddhist.The prevalence of diffuse goiter, nodular goiter, colloid goiter, thyroid adenoma, thyroid carcinoma, hyperthyroidism, subclinical hyperthyroidism, hypothyroidism, subclinical hypothyroidism, and positve rate of TPOAb were 1.7% ,25.3% ,8.7% ,0.2% ,0.4% ,0.5% ,0.8% ,0.03%,1.0% ,and 9.5% repectively.The prevalence of thyroid diseases was increasing with aging, and higher in women than in men (P＜0.05).There was no significant relationship of the thyroid diseases with seafood, smoking,drinking, and tea (P＞0.05).Conclusions The citizens of Zhoushan archipelago have adequate iodine intake.It is pertinent to discuss Universal Salt Iodization.Excessive iodine intake may contribute to the high prevalence rate of thyroid diseases in Zhoushan.

OBJECTIVETo investigate the apoptosis of hepatocytes and the expression of apoptosis-regulating genes during the donor liver ischemia and reperfusion injury in rat orthotopic liver transplantation (OLT).

METHODSSeventy-two male SD rats were randomly divided into sham operation group and transplantation group. Using Ringer's lactate solution as the perfusing and preserving solution, the grafts were preserved for 4 h before orthotopic transplantation. At 1, 6 and 24 h after the reperfusion, the recipients were sacrificed, and the serum ALT and AST levels were measured; the changes of hepatocyte apoptosis was detected by TUNEL assay, and the protein expressions of the apoptosis-regulating genes were measured by flow cytometry.

RESULTSSerum ALT and AST levels were significantly higher in transplantation group than in the control group after reperfusion. In comparison with the control group, the rats in the transplantation group showed significantly increased apoptosis index in the livers, lowered Bcl-2 levels and increased FasL levels after the transplantation, especially at 6 h after liver reperfusion (P<0.01).

CONCLUSIONThe donor liver ischemia and reperfusion injury can promote hepatocyte apoptosis, which may be related with the high expression of Bcl-2 gene and low expression of FasL after reperfusion injury in rats with orthotopic liver transplantation.

Animals ; Apoptosis ; Fas Ligand Protein ; metabolism ; Gene Expression Regulation ; Hepatocytes ; cytology ; metabolism ; Liver ; metabolism ; Liver Transplantation ; adverse effects ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology