Objective To investigate the serum proteomic patterns in lung cancer by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) techniques and build diagnostic models in order to evaluate their clinical significance by biomarkers screening in lung cancer.Methods SELDI-TOF-MS and CM-10 protein chip were used to detect the serum proteomic patterns of 38 lung squamous cell carcinoma and 30 lung adenocarcinomas,including the comparation in 34 geminate patient serums before and after surgery.Nagative control setted as a group of 50 normal tissues.BioMarker Wizard and BioMarker pattern system software were used in combination to analyze the data and to develop diagnostic models.Results Two protein peaks from a total of 167 were chosen as a biomarker pattern in the training set.Two protein peaks pattern (mass/charge ratio [m/z] 6 010,5 330) was observed in the model that could be used as to distinguish lung cancer from non-cancerous diseases.It yielded a sensitivity of 98 ％ and a specificity of 96 ％.There were 9 different protein peaks (P ＜ 0.05) between pre-surgery and post-surgery.There were 8 different peaks (P ＜ 0.05) between metastasis and non-metastasis.Conclusion SELDI techniques can be employed as diagnosis in lung cancer patients with relatively high sensitivity and specificity,and also could be used as an effective tool for the screening of lung cancer.

microRNA with 22 nucleotides in length are a growing family of small non-coding RNA that function as post-transcriptional regulators of target genes in eukaryote.Recent study indicates that microRNA are closely related to the oncogenesis additional to its biological function in the regulation of cell proliferation,differentiation and death.This review proposes the progresses on the study of(microRNA,) including the biological character,biogenesis and function,and its relationship with cancer.

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.

Objective To establish acute peritonitis induced by monosodium urate (MSU) of in mice and observe the significance of mitochondrial deoxyribonucleic acid (mtDNA) expression in the inflammatory processes.Methods The mouse models of acute peritonitis were made by intraperitoneal injection of MSU.Sixty-four male C57BL16 mice were randomly divided into the MSU group which were treated with 0.2 ml of 15 mg/ml MSU solution by i.p.injection and the control group which were treated with 0.2 ml of PBS.Respectively four mice from MSU group and four mice from control group were killed 2 hours, 4 hours, 6 hours, 8 hours 12 hour, 16 hours, 20 hours and 24 hours later and whole blood, peritoneal lavage and peritoneum were collected respectively.Four the mice from the MSU group and four mice from the control group were killed and whole blood, peritoneal lavage and peritoneum were collected.Immunoflourescence study of peritoneum tissues was performed.The levels of interleukin (IL)-1β, IL-18 in plasma and peritoneal lavage were examined by enzyme linked immunosorbent assay (ELISA).DNA was extracted from blood and peritoneal lavage, and mtDNA level was detected by using real-time polymerase chain reaction (PCR).The data was analyszied by multivariate analysis of variance.Results As compared with those killed at other time points from the MSU groups and the control group, the levels of IL-1β [(27.0±2.0) pg/ml vs (26.8±2.1) pg/ml], IL-18 [(673±454) pg/ml vs(752±495) pg/ml] in plasma and peritoneal lavage were increased progressively in those which were killed after i.p.injection of 2 hours and 4 hours from in the MSU group (F=22.778, P＜0.05;F=6.660, P＜0.05).The mtDNA in plasma and peritoneal lavage of the mice began to be expressed 4 hours after i.p.injection 4 hours from in the MSU group.The peak level was detected in those i.p.injected MSU 6 hours later [(9.85±4.59)×106 copies, (7.81±3.43)×106 copies].Then 8 hours later the mtDNA began to slowly decreased.At these three time points, the mtDNA were all increased progressively than those at the other time points of the MSN group or at all time points of the control group (F=6.719, P＜0.05;F=11.181, P＜0.05).By immunoflourescence study, there were neutrophil extracellular traps (NETs) were formed 12 hours later in the MSU group and aggregated NETs were found 24 hours later.Conclusion In the inflammatory processes of acute peritonitis induced by MSU of in mice, with the expression of mtDNA increasing, the inflammation is relieved, and aggregated NETs are formed in the end.Expression of mtDNA may be one for the protective factors of the inflammation induced by MSU.

Objective To analyze the efficacy and hemodynamic changes in patients with complex symptomatic middle cerebral artery stenosis after Solitaire AB stent implantation.Methods From June 2013 to June 2016,7 consecutive patients with complex symptomatic middle cerebral artery stenosis treated with Solitaire stents at the Department of Neurology,the Third People's Hospital of Hubei Province were enrolled retrospectively.The stenosis rate ≥70% was confirmed by digital subtraction angiography (DSA).Of the 7 patients,5 were male and 2 were female,with an average age of 70±8 years.The general data of the patients were documented,including clinical symptoms,modified Rankin scale(mRS)score,stenosis degree,vascular pathways LMA classification and Mori typing for stenosis,and postoperative residual stenosis or restenosis.The hemodynamic parameters at day 1,7,and 3 months before and after procedure were compared.Results All patients were successfully implanted with the Solitaire AB stents.(1) Of the 7 patients,the Mori type A of stenosis lesions was in 4 cases,B in 2 cases,and C in 1 case.The LMA typing of vascular approaches were Type Ⅲ in 3 cases,type Ⅱ in 4 cases.The preoperative stenosis rate was 80±7%,the residual stenosis rate was 24±13%,and there were no operation-related complications.No ischemic events occurred during the follow-up period.(2) Before stent implantation and at 1 d,7 d,and 3 months after procedure,the peak systolic velocity (PSV) were 297±41,144±34,145±27,and 143±40 cm/s,respectively,the end diastolic velocity (EDV) were 159±22,68±16,68±14,and 66±17 cm/s,respectively,and the pulse index (PI) were 0.67±0.06,0.80±0.08,0.80±0.07,and 0.82±0.06,respectively.PSV,EDV,and PI at 1 d,7 d,and 3 months were improved compared with before procedure (all P<0.05),There were no significant differences at all time points in the pairwise comparison PSV and EDV and PI at all time points after the procedure (all P>0.05).Conclusions Initial analysis showed that no procedure-related complications were observed in the treatment of vascular tortuosity and complex middle cerebral artery stenosis with the Solitaire stents.The short-term curative effect is exact;However,it still needs to be validated further.

Objective:To investigate the association of FAT atypical cadherin 1 (FAT1) with clinicopathological parameters and prognosis in esophageal squamous cell carcinoma (ESCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 124 patients with ESCC who were admitted to Shanxi Cancer Hospital from January 2011 to December 2015 were collected. There were 85 males and 39 females, aged from 40 to 72 years, with a median age of 60 years. The ESCC tissues surgically removed and adjacent tissues specimens were collected to prepare tissue microarray for immunohistochemical staining. The 5 cases of ESCC tissues and adjacent tissues were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Observation indicators: (1) the expression of FAT1 protein in ESCC and adjacent tissues; (2) the expression of FAT1 RNA in ESCC and adjacent tissues; (3) the expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters; (4) follow-up and survival. Follow-up using outpatient examination and telephone interview was conducted to detect survival of patients up to February 13, 2019. The survival time was from surgical date to tumor-related death or endpoint of follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the non parameter rank sum test. The Kaplan-Meier method was used to calculate survival time, and Log-rank test was used for survival analysis. Results:(1) The expression of FAT1 protein in ESCC and adjacent tissues: of 124 specimens, the 107 cases of ESCC tissues and 93 cases of adjacent tissues were finally obtained because of exfoliative tissues. There were 76 cases of ESCC tissues and corresponding adjacent tissues matched. Results of immuno-histochemical staining showed that FAT1 protein was expressed in both ESCC and adjacent tissues and was brown after staining. FAT1 was located in cytomembrane, with high expression of FAT1 as ≥75 and low expression as <75. The relative expression levels of FAT1 protein in ESCC and adjacent tissues were 68±42 and 77±37, showing a significant difference between ESCC and adjacent tissues ( t=2.380, P<0.05). (2) The expression of FAT1 RNA in ESCC and adjacent tissues: results of qRT-PCR showed that the relative expression levels of FAT1 RNA in 5 cases of ESCC and adjacent tissues were 1.6±0.4 and 2.5±0.3, with a significant difference between them ( t=3.560, P<0.05). (3) The expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters: of the 107 ESCC patients, 58 cases had high expression of FAT1. There were 42 and 16 cases with high expression of FAT1 in 65 non-drinking patients and 42 drinking patients, respectively, showing a significant difference between them ( χ2=7.229, P<0.05). (4) Follow-up and survival: 96 of 107 ESCC patients were followed up for 38.0?94.9 months, with a median follow-up time of 45.9 months. Survival analysis showed that the survival time of patients with high FAT1 expression was 24 months, versus 22 months of patients with low FAT1 expression, indicating no significant difference between them ( χ2=1.773, P>0.05). Results of subgroup analysis showed that the survival time was 24 months and 21 months of female patients with high and low FAT1 expression, 23 months and 22 months of non-smoking patients with high FAT1 expression and low FAT1 expression, 23 months and 21 months of non-drinking patients with high FAT1 expression and low FAT1 expression, respectively, showing significant differences between them ( χ2=8.769, 12.827, 10.724, P<0.05). Conclusions:The expression of FAT1 in ESCC tissues is low. Female, non-smoking and non-drinking ESCC patients with high FAT1 expression have good survival.

Objective To investigate the expressions of Th17 lymphocytes and interleukin-17 (IL-17) in peripheral blood of patients with non-small cell lung cancer (NSCLC) and its clinical significance. Methods Sixty patients with primary and untreated NSCLC were enrolled and designed as experimental group, at the same time, 60 healthy volunteers were collected as control group. Flow cytometry (FCM) was used to detect the level of Th17 lymphocytes. Enzyme linked immunosorbent assay (ELISA) was used for detecting the level of IL-17. The relationship between the expression levels of Th17 and IL-17 in peripheral blood and clinicopathological features was compared between the two groups. Results The peripheral blood levels of Th17 lymphocytes and IL-17 in the experimental group [(1.7±1.2) ％, (8.3±2.5) pg/ml] were higher than those in the control group [(0.9 ±0.6) ％, (5.4 ±1.2) pg/ml] (P< 0.05). The peripheral blood expression of Th17 lymphocytes and IL-17 in patients with smoking history [(1.8±1.2) ％, (8.8±3.7) pg/ml] were higher than those in patients without smoking history [(1.6±1.2)％, (8.0±2.2) pg/ml], and the peripheral blood expression of Th17 lymphocytes and IL-17 were higher in patients with squamous-cell carcinoma [(1.8 ±1.2) ％, (9.4 ±4.7) pg/ml] than those in patients with adenocarcinoma [(1.6±1.1) ％, (7.3±3.9) pg/ml], furthermore, they were also higher in patients with stage Ⅲ-Ⅳ than those in patients with stage Ⅰ-Ⅱ (P < 0.05). Conclusion Th17 lymphocytes and IL-17 play certain roles in the occurrence and progression of NSCLC.

Objective:To explore the practical effect of multi-disciplinary treatment learning (MDT-L) combined with PBL+CBL teaching on the clinical clerkship of neurosurgery.Methods:In the study, 30 neurosurgery clinical interns accepted by Wuhan Third Hospital from January 2019 to December 2019 were set as the control group, and 30 neurosurgery clinical interns from January 2020 to December 2020 were set as the research group. The control group implemented conventional teaching, and the research group used MDT-L combined with PBL+CBL teaching. After the clerkship, the two groups' clinical clerkship results, changes of comprehensive abilities before and after clerkship, professional quality after clerkship, and satisfaction with teaching methods were compared. SPSS 26.0 was used for t test and chi-square test. Results:After the clerkship, the theoretical knowledge and operational skills assessment scores of the research group were significantly higher than those of the control group ( P<0.05). There was no statistically significant difference in the scores of the two groups in learning interest, analytical ability, communication skills, innovation ability and file management ability before the clerkship ( P>0.05). While, after the clerkship, the comprehensive ability scores of the above dimensions of the two groups increased, and the comprehensive ability scores of the study group were all higher than those of the control group ( P<0.05). After the clerkship, the scores of professional ability, problem analysis and solving ability, mastery of diagnosis and treatment procedures, scoring of medical documents writing and total scoring of professional quality in the study group were all higher than those in the control group ( P<0.05). The satisfaction scores of the study group on the attractiveness, effectiveness and practicality of the teaching method were higher than those of the control group ( P<0.05). Conclusion:The application of MDT-L combined with PBL+CBL teaching in neurosurgery clinical clerkship can not only improve the performance of internship, but also enhance the comprehensive ability of interns and improve their professional quality and satisfaction.

Objective:To investigate the expression of miRNA-34b (miR-34b) in non-small cell lung cancer (NSCLC) tissues and its effect on proliferation and invasion of human NSCLC A549 cells in vitro.Methods:The specimens of cancer tissues and paracancerous normal epithelial tissues (more than 5 cm from the edge of the tumor) were collected from 40 NSCLC patients in Shanxi Province Cancer Hospital from June 2015 to March 2017. A549 cells were transfected with miR-34b mimics (experimental group) and irrelevant sequences (negative control group), respectively. The expression of miR-34b in tissues and each group of A549 cells was detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The proliferation activity of A549 cells in the experimental group and the negative control group was detected by methyl thiazolyl tetrazolium (MTT) assay, and the invasion ability of A549 cells in the two groups was detected by Transwell assay.Results:The relative expression of miR-34b in NSCLC tissues was lower than that in paracancerous normal epithelial tissues (0.52±0.06 vs. 1.05±0.17), and the difference was statistically significant ( P < 0.001). The relative expression of miR-34b in A549 cells of the experimental group was higher than that in the negative control group, and the difference was statistically significant ( P < 0.05). MTT assay showed that the cell proliferation ability (absorbance value) of A549 cells in the experimental group was lower than that in the negative control group after cultured for 24 and 48 hours (both P < 0.01). Transwell assay showed that the number of invaded A549 cells in the experimental group was less than that in the negative control group [(49.53±5.03) cells vs. (121.00±12.06) cells, P < 0.01]. Conclusions:The expression of miR-34b is low in NSCLC tissues, and the up-regulation of miR-34b expression can inhibit the proliferation and invasion of NSCLC A549 cells.