Objective To evaluate the anti-tumor effects of chemotherapeutic drugs on human gastric cancer cells. Methods From April 2006 to February 2007, 84 patients with gastric cancer underwent surgical resection at General Hospital of Jinan Military Command. The single-cell suspension of these gastric tumors was prepared. The gastric cancer cells were cultured with hydroxycamptothecin, cisplatin, adriamycin, 5-fluorouracil and mitomycin for 48 hours, and changes in activity of the gastric cancer cells were studied via MTT assay. The expression of survivin and PTEN was detected by immunohistochemistry. Data were analyzed by chi-square test, rank sum test and Fisher exact test. Results The anti-tumor effects of different chemotherapeutic drugs were different, and the poorly-differentiated gastric cancer cells were more sensitive to the cytotoxic effects of chemotherapeutic drugs than the well-differentiated gastric cancer cells. The expression levels of survivin in the signet ring cell carcinoma, mucinous adenocarcinoma and other poorly-differentiated adenocarcinoma were significantly higher than those in papillary carcinoma and tubular carcinoma (χ~2 = 10.625, P <0.05), while the expression of PTEN was inverse to that of survivin (χ~2 = 6.060, P < 0.05). The expression of survivin was related to the resistance of the gastric cancer cells to 5-fluorouracil and adriamycin (χ~2 = 6.609, 6.350, P < 0.05). Conclusions In vitro chemosensitivity assay is helpful in selecting the chemotherapeutic regimen for specific types of gastric cancer. Survivin may contribute to the chemotherapy-resistance of certain types of gastric cancer cells, and its expression is related to that of PTEN.

Objective To investigate the expression of Wnt-5a gene in primary hepatocellular carcinoma (HCC) and to expose its role and clinical significance in the development of HCC.Methods Real time quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed in 26 fresh HCC samples and the corresponding para-carcinoma tissues to detect mRNA expression of Wnt-5a gene.Wnt-5a protein was detected with immunohistochemical method in paraffin embedding tissues of 85 cases of HCCs and the corresponding para-carcinoma tissues,and 15 cases of hepatic cirrhosis.Results RT-PCR analysis showed that Wnt-5a mRNA (0.102 127 ±0.158 620) in the HCC tissues was more than that (0.020 106 ±0.022 075) in the para-carcinoma tissues (P＜0.05).The positive expression rate of Wnt-5a protein in HCC,para-carcinoma,and hepatic cirrhosis tissues were 21.2％ (18/85),81.26％ (69/85),and 86.7％ (13/15),respectively.The positive rate of Wnt-5a was significantly lower in the HCC than in the para-carcinoma and hepatic cirrhosis tissues (P ＜ 0.01).The expression of Wnt-5a was significantly associated with lower tumor node metastasis (TNM) stages and small alpha fetoproteins (AFP) content of blood serum (P ＜0.05).Conclusions The high expression of Wnt-5a mRNA was found in the gene transcription of HCC,while Wnt-5a protein was absent or low in HCC.It was suggested that the roles of Wnt-5a was interfered at the protein level rather than the transcriptional level in the HCC.

OBJECTIVETo observe the pathological changes in rabbits with spinal cord injury induced by decompression sickness (DCS), and to investigate the role of tumor necrosis factor-alpha (TNF-α) in spinal cord injury induced by DCS.

METHODSRabbits were randomly divided into normal control group, DCS group, and safe decompression group. The rabbit model of DCS was established. Light microscopy, real-time PCR, and immunohistochemical method were used to observe the pathomorphological changes in the thoracolumbar spinal cord and the mRNA and protein expression of TNF-α, respectively. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to observe the apoptosis in the spinal cord.

RESULTSIn the DCS group, cavities formed in the white matter of spinal cord and gliosis occurred around necrotic areas. Moreover, the mRNA and protein expression of TNF-α was significantly higher in the DCS group than in the normal control group and the safe decompression group (P<0.01). The results of TUNEL showed that the number of positive apoptotic cells was significantly larger in the DCS group than in the normal control group and the safe decompression group (P<0.05).

CONCLUSIONApoptosis plays an important role in spinal cord injury induced by DCS. In the early stage of DCS, the massive release of TNF-α initiates apoptosis and contributes to the pathological changes in spinal cord injury induced by DCS.

Animals ; Apoptosis ; Decompression Sickness ; metabolism ; pathology ; Disease Models, Animal ; In Situ Nick-End Labeling ; RNA, Messenger ; Rabbits ; Spinal Cord ; pathology ; Spinal Cord Injuries ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the effects of chemotherapeutic drugs on ER-α expression and methylation in breast cancer cells.Methods Human breast cancer cells MCF-7(ER+,Luminal A) were induced by paclitaxel(PTX) and epirubicin(EPI) for more than 6 months,with an incremental dose,respectively.The expression and methylation status of ER-α in MCF-7 cells were detected before and after drug treatment.miRNAs with consistent expression changes in MCF-7 cells after two drugs' treatment were screened by microarray,and verified by quantitative PCR (qPCR).Targets of the most significantly down-regulated miRNA were analyzed by bioinformatics.miRNA inhibitor was transfected into MCF-7 cells,miRNA mimic was transfected into MCF-7/PTX and MCF-7/EPI cells,then ER-α and DNA methyltransferase 1 (DNMT1) expression were detected by Western blot,and ER-α methylation was detected by quantitative methylation-specific PCR (qMSP).Results PTX resistant MCF-7/PTX cell line and EPI resistant MCF-7/EPI cell line were established.Both drug treatments caused a decrease in ER-α protein expression and an increase in methylation levels,with up-regulation of DNMT1 and his tone deacetylase 1 (HDAC 1) expression.miRNAs with consistent expression changes in MCF-7 cells after drug treatments were screened and verified by qPCR,the most significant down-regulation among which was miR-148b.Bioinformatics analysis,and further confirmed by luciferase reporter gene assay (Luciferas) that DNMT1 was a direct target of miR-148b.miR-148b inhibitor induced decreased expression of ER-α and increased methylation level in MCF-7 cells,accompanied by increased expression of DNMT1;whereas miR-148b mimic caused an increased expression of ER-α and decreased methylation level in MCF-7/PTX and MCF-7/EPI cells,with a decreased expression of DNMT1.Conclusion Chemotherapeutic drugs (represented by PTX and EPI) induce aberrant miRNA expression in breast cancer MCF-7 cells,and down-regulate miR-148b further to attenuate the inhibition of DNMT1 expression,which promote,hypermethylation and down-regulation of ER-α.

Objective To study the sero-epidemiology of pertussis immunity and related factors among the community-based populations in Tianjin.Methods Antibodies to pertussis toxin immunoglobulin and IgG (PT-IgG) levels among the community-based populations were examined,using ELISA over the three consecutive years.Subjects that were older than 4 years of age,with concentration of anti-PT-IgG higher than 40 IU/ml were recognized as having recent Bordetella (B.) pertussis infection.Results Of the 1 825 study subjects,the average positive rate of anti-PT-IgG was 10.96％.The highest rate appeared as 24.37％-13.61％ among the 0-3 year-olds (P＜0.001).The positive rate was 8.84％,and the estimated incidence of recent infection became 10 852 per 100 000 among those age 4-83 year-olds.Peak of the estimated incidence rate of recent infection was 18 986 per 100 000 in the age group of 51-83 year-olds (P=0.001),but increasing linearly along with the increase of age (r=0.976,P＜0.001).There were statistically significant differences seen in the antibody positive rate of 3-year period of study (P=0.001),and appeared linear correlation with the reported annual incidence (r =0.992,P＜ 0.001).There were statistically significant differences in the antibody positive rates of the 3 monitoring areas (P=0.034),also showing a linear correlation with the reported annual incidence (r=0.996,P＜0.001).Conclusion Results from this study indicated that B.pertussis infection had been common,particularly in adults living in the communities in Tianjin,calling for the current pertussis immunization strategy to be improved in order to control the pertussis reemerge in China.

Objective:To analyze the clinical characteristics of Chlamydia psittaci pneumonia in the elderly, and factor related to the disease severity. Methods:Clinical data of 32 elderly patients with Chlamydia pneumoniae pneumonia admitted in the First People′s Hospital of Xiaoshan District from January 2019 to January 2021 were retrospectively analyzed. The patients were diagnosed by the second generation sequencing using bronchoalveolar lavage fluid (BALF) samples. There were 17 patients in moderate group and 15 patients in severe group; the liver function, muscle enzymes, imaging and lymphocyte subsets of the two groups were compared. Results:There were no significant differences in the age, gender and basic diseases, bird contact history, flaccid fever and dry cough symptoms between two groups, while there were significant differences in mental symptoms and gastrointestinal symptoms between the two groups ( P=0.032, 0.018). There were significant differences in ALT , AST, LDH , CK , CK-MB, CRP, BNP , troponin-T and PCT between the two groups (all P<0.001). The increase of leukocytes was not significant in both groups. The moderate group was dominated by single lobe involvement, while the severe group was dominated by multi lobe involvement and pleural effusion ( P=0.043, 0.015, 0.023). The total lymphocytes, T lymphocytes, CD4 + T cells, CD8 + T cells, CD4 +/CD8 + ratio, NK cells, B cells, CD4 +CD8 + double positive T cells decreased in both groups, while those in the severe group decrease more markedly ( P<0.05). CD4 -CD8 - double negative T cells were higher in the severe group than those in the moderate group ( P<0.001). CD4 +/CD8 + ratio and CD4 -CD8 - double negative T cells were correlated with severity index PSI and CURB-65 ( P<0.05). Conclusions:The liver function, muscle enzyme, lymphocyte immune function in patients with Chlamydia psittaci pneumonia are impaired, which were more markedly in severe patients. The multileaf infiltration and increased procalcitonin may indicate the severe pneumonia. CD4 +/CD8 + ratio and CD4 -CD8 - double negative T cells are correlated with the severity of pneumonia.

OBJECTIVETo investigate the changes in expression of tumor necrosis factor-alpha (TNF-α) and glial fibrillary acidic protein (GFAP) in rabbits with decompression disease (DCS), and to investigate the functioning mechanism.

METHODSA total of 21 healthy adult rabbits were randomly divided into 3 groups: normal control group, DCS group, and safe relief group, with 7 rabbits in each group. A rabbit DCS model was established by quick decompression. The changes in pathological morphology and mRNA and protein expression of TNF-α and GFAP in the brain and spinal cord of rabbits with DCS were determined by light microscopy, real-time PCR, and immunohistochemistry, respectively.

RESULTSCavity formation was observed in the white matter of spinal cord in DCS group. The mRNA and protein expression of TNF-α and GFAP was significantly higher in the DCS group than in the normal control group and safe relief group (P < 0.01), while no significant differences were observed in the brain (P > 0.05).

CONCLUSIONSpinal cord is the main part of central nervous system injury in DCS. Activation of TNF-α and GFAP genes accompanied by increase in their protein expression can be observed at the early stage of DCS. The astrocytes and TNF-α play important roles in the process of spinal cord injury in DCS.

Animals ; Brain ; metabolism ; Decompression Sickness ; metabolism ; Disease Models, Animal ; Glial Fibrillary Acidic Protein ; metabolism ; Male ; Rabbits ; Spinal Cord ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo evaluate the application of mismatch repair (MMR) genes proteins expression to screen for Lynch syndrome in colorectal cancer patients.

METHODSOne hundred consecutive colorectal cancers cases collected from 2012 to 2013 were tested immunohistochemically for the protein expression of MLH1, MSH2, MSH6 and PMS2, and also by the ARMS method for the mutation status of BRAF genes in those cases lacking protein expression for MLH1.

RESULTSThe result of MMR immunocytochemistry showed that nine of 100 cases lacked MMR protein expression, including three cases each that were MLH1-/PMS2- and MSH2-/MSH6- respectively, two cases were MLH6- and one case was PMS2-; overall, the majority of these cases lacked protein expression of MLH1 and MSH2. The BRAF genes mutation test showed one case of mutation, indicating that the patient might have MLH1 gene methylation as a result of the mutation of BRAF genes, and that was a sporadic case. The age of onset for patients lacking MMR protein expression was lower than patients with sporadic colorectal cancer (P = 0.011). Colorectal cancers associated with the lack of MMR protein expression mostly occurred in the right colon (P = 0.001), and histomorphologically were often accompanied by mucinous adenocarcinoma (P = 0.010) and tumor lymphocytic infiltration.

CONCLUSIONImmunohistochemical staining for MMR proteins in patients with colorectal cancer, accompanied by testing for BRAF genes mutation, may be an effective approach to screen for Lynch syndrome.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Colorectal Neoplasms, Hereditary Nonpolyposis ; diagnosis ; genetics ; DNA Mismatch Repair ; Humans ; Immunohistochemistry ; MutL Protein Homolog 1 ; Mutation ; Nuclear Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism