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Objective To study the pathological changes and expressions of NO and iNOS mRNA in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the lung injury.Methods A total of 140 male SD rats were randomly (random number) ivided into the room temperature (25 ℃) environment traumatic hemorrhagic shock group (room temperature group) and the dry heat traumatic hemorrhagic shock groups (dry heat group,temperature 40℃,humidity 10％),respectively,and each groups was further randomly divided into 7 subgroups:the control subgroup,post shock subgroups at 0,0.5,1,1.5,2and 3 h (n =10 in each subgroup).The rats of control subgroup were not treated,and rats of dry heat group were placed in dry heat environment for 60 min,then anesthetized,fixed,and insertion of intravenous indwelling needles and catherization of right carotid artery,jugular vein and the right femoral artery were performed.After stabilization for 10 min,2500 g iron wheel was used to be dropped from 30 m height and vertically hit the upper left femoral of SD rats in order to make comminuted fracture,wounds were quickly dressed after injury.Exsanguination from right femoral artery was kept until MAP maintained at (35 ± 5) mmHg,and resuscitation was carried out after continue monitoring for 60 min.After the establishment of traumatic hemorrhagic shock model in each environment,the rats were sacrificed at given intervals,and thoracotomy was performed to take broncho-alveolar lavage fluid (BALF) and lung tissue.Pathological changes of lung tissues were observed by using HE staining and NO concentration of lung tissue was detected by one-step method,and changes of the iNOS mRNA expressions were detected by using fluorescence quantitative PCR.Then t test,ANOVA and Pearson correlation analysis were used for the data analysis.Results The pathological change in dry heat group at each interval was more severe,and pulmonary histopathological injury score was higher,and the protein exudation was more profuse compared with the room temperature group.NO concentration in lung tissue homogenate of dry heat group was higher than that of room temperature group (t =2.472,P ＜ 0.05),and the difference in NO level between different intervals within the dry heat group was statistically significant (F =6.77,P ＜ 0.01).The NO concentration in dry heat group reached its maximum at 2 h (3.35 ± 0.23) μmol / g and the peak value emerged sooner than that in room temperature group.The difference was statistically significant in overall expression of iNOS mRNA between two groups analyzed with t test (t =3.619,P ＜ 0.01),and there was statistically significant difference between intervals within the dry heat group (F =12.34,P ＜0.01).The values of iNOS mRNA in the dry heat group were higher than those in the room temperature group at the same given intervals,and the peak value appears at 1.5 h in dry heat group,and the room temperature group it began to increase at 2 h.The concentration of NO and the expression of iNOS mRNA were positively correlated with each other in two groups (r =0.680,r =0.376).The expression of iNOS mRNA and lung histopathological injury score was positively correlated in two groups (r =0.846,r =0.899).Conclusions When traumatic hemorrhagic shock occurred in the dry heat desert environment,the lung injury was more severe and appeared sooner than that in the room temperature environment.NO and iNOS played important roles in the secondary lung injury in the wake of traumatic hemorrhagic shock in rats under the dry heat environmengt of desert.
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cryopreservation periodontal tissue. METHODS:Periodontal tissue was scraped from healthy human teeth and divided them into three equal parts:fresh group, harvesting periodontal ligament stem cel s from fresh tissue;5%dimethyl sulfoxide group, 5%dimethyl sulfoxide added into the cryopreservation system;10%dimethyl sulfoxide group, 10%dimethyl sulfoxide added into the cryopreservation system. One month later, periodontal ligament stem cel s were extracted from the cryopreserved periodontal ligament from the latter two groups. RESULTS AND CONCLUSION:In the time that cel s swam out of tissue mass and cel harvest amount, the 5%dimethyl sulfoxide group was inferior to the fresh group but better than the 10%dimethyl sulfoxide group (P0.05):colony formation rate of passage 1 periodontal ligament stem cel s, cel survival rate, proliferation ability of passage 3 periodontal ligament stem cel s, cel growth curve and surface marker expression of periodontal ligament stem cel s. The results suggest that the 5%dimethyl sulfoxide added cryopreserved system for periodontal ligament tissue cannot only shorten the time of periodontal ligament stem cel s amplification in vitro, ensure cel harvest and maintain basic cel ular biological characteristics, but also reduce the total amount of the dimethyl sulfoxide and the direct damage to the cel s caused by repeatedly frozen cel s, thereby providing a more secure guarantee for the future implementation of clinical transplantation therapy. So, the 5%dimethyl sulfoxide added cryopreserved system may be the new selection for donor tissue storage.
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Objective To investigate the role of microRNA-21 (miRNA-21) in promoting proliferation of Kazakh's esophageal squamous cell carcinoma (ESCC) cell line Eca109 and Kazakh's ESCC tissues, and its effects on the expression of programmed cell death 4(PDCD4)gene.Methods TheEca109cellsweredividedintomiRNA-21Mimicsgroup(transfersequence:5′-UCAACAUCAGUCUGAUAAGCUA-3′),miRNA-21inhibitorgroup(transfersequence:5′-UAGCUUAUCAGACUGAUGUUGA-3′), negative control group (the transfer random sequence)and normal control group (no transfection).The cells were seeded at 4.5 × 105/well in 6-well cell culture plates. According to RNA interference technology,thecells were transfected with LipofectamineTM 2000. After transfection, the cell proliferation was determined by cell number counting.The expression of miRNA-21 was detected by real-time quantitative polymerase chain reaction (qRT-PCR), and the expression of PDCD4 protein was measured by Western blot.Meanwhile, the expression of miRNA-21 and PDCD4 protein were performed in 18 pairs of Kazakh's ESCC and corresponding adjacent normal esophageal mucosa.ResultsAt 48 hour after transfection,compared with normal control group, the cell number increased 36％ (P=0.002) in miRNA-21 Mimics group, decreased 28％ (P=0.002) in miRNA-21 inhibitor group, and did not change significantly in negative control group (P=0.515). At 48 hour after transfection, the relative expression of miRNA-21 in miRNA-21 Mimics group, miRNA-21 inhibitor group, negative control group and normal controlgroup was 0.37±0.10, 9.17± 1.08, 0.74±0.23 and 1.04±0.34,respectively, and the relative protein expression of PDCD4 was 1.47 ± 0.11, 0.61±0.09, 0.89 ±0.12 and 0.79±0.02 accordingly.Compared with normal control group, the relative expression of miRNA-21 decreased (P=0.031) and the relative protein expression of PDCD4 up regulated (P=0.001) in miRNA-21 inhibitor group, the relative expression of miRNA-21 increased (P=0.001) and the relative protein expression of PDCD4 down regulated (P=0.030) in miRNA-21 Mimics group,and there was no significant difference in negative control group (P=0.272 and 0.541).In 16 pairs of Kazakh's ESCC tissues, the expression of miRNA-21 was significantly higher in tumor tissues (0.11 ±0.09) than that of corresponding adjacent esophageal normal tissues (0.03 ± 0.03, P=0.001), while the relative protein expression of PDCD4 was significantly lower (0.92 ± 0.39) than that of corresponding adjacent normal esophageal tissues (1.57 ± 0.80, P=0.004). And the higher expression of miRNA-21, the lower relative protein expression of PDCD4 (r=-0.538, P=0.046).ConclusionmiRNA-21 may promoted the cell proliferation through inhibiting PDCD4 expression,which involved in the pathogenesis of Kazakh's ESCC.
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Objective To explore the relationship between polymorphism of catechol-O-methyltransferase (COMT) gene valine (Val) 158 methionine (Met) (G to A transition)and the distribution in population and esophageal squamous cell carcinoma (ESCC) in Yili prefecture of Xinjiang.Methods A hospital based case-control study was adopted, a total of 622 subjects, which including 214 ESCC patients and 408 age, gender and ethnicity-matched normal control individuals.The polymorphism of COMT gene G to A transition was analyzed with PCR-restriction fragment length polymorphism approaches.Results The COMT genotype frequencies in 622 subjects in Yili prefecture were GG genotype accounted for 47.3%, GA type for 42.3% and AA type for 10.4%, G allele was 68.4% and A allele was 31.6%.There was no statistical difference in the COMT genotype and frequencies of allele distribution between ESCC group and control group.Furthermore, stratified analysis indicated that there was statistical difference between ESCC group and control group in subjects less than 60 years old.There was statistical difference in the allele distribution among Kazak,Uygur and Han ESCC groups.The COMT genotype and frequency of allele distribution among normal control groups of the three ethnic groups were statistically different.After corrected age and gender,there was no statistical difference in COMT Val158Met polymorphisms among Kazakh, Uygur and Han ethnic groups in both ESCC and control groups in Yili Prefecture of Xinjiang.Conclusion COMT gene Val158Met single nucleotide polymorphism may not be the genetic markers of ESCC risk in Yili Prefecture of Xinjiang.
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Objective To estimate the effect of microRNA (miRNA) let-7 expression on human esophageal squamous cell carcinoma(ESCC) and the relationship between let-7 level and clinicopathological parameters. Methods ESCC cell line (Eca109) was transfected with let-7 or its inhibitor by RNAi and cell transfection techniques. Normal cultured Eca109 cell was served as negative control. The proliferation of Eca109 cell was detected by MTT. The expression of let-7 in Eca109 cells and 45 paired ESCC tissues and corresponding para-cancerous tissues were measured using real-time quantitative polymerase chain reaction (qRT-PCR). The relationship between let-7 level and clinicopathological parameters in patients with ESCC was analyzed. Results The A value of let-7 in Eca109 cells transfected with let-7 was lower than negative control (P=0.005), while it was higher in Eca109 cells transfected inhibitor than that in negative control 72 hours after transfection. In comparison with negative control, the expression of let-7 in Eca109 cells transfected with let-7 was increased 33% (1.33 vs 1.00,P=0. 039) and it was decreased 50% in Eca109 cells transfected with inhibitor (0.50 vs 1.00,P=0. 014). The ratio of let-7 expression in ESCC tissue and para-cancerous tissue was 0.66 ± 0.47 with significant differece (P= 0.001). Moreover, The level of let-7 expression in Han patients with ESCC was lower than Kazakh patients with ESCC (0.48±0.43 vs 0. 88±0.51,P=0. 019). The level of let-7 expression in poorly differentiated ESCC tissue was lower than well differentiated ESCC tissue (0.42±0.30 vs 0.84±0.38,P=0. 015). The level of let-7 expression in patients with lymph node metastasis was lower than those without lymph node metastasis (0.50±0.35vs 0. 80±0.52,P=0. 032) . Conclusion It is demonstrated that let-7 can inhibit the carcinogenesis and development of ESCC. The level of let-7 expression is associated with cell differentiation,lymph node metastasis and nationalities.
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Objective To explore Annexin A2 expression in human esophageal squamous cell carcinoma (ESCC) and investigate the correlation of Annexin A2 expression with invasion and metastasis of human ESCC. Methods From 2000 to 2008, specimens of Xinjiang medical University First Affiliated Hospital were collected. Pathologically confirmed ESCC surgical specimens were set as experimental group, and the corresponding tumor adjacent tissues located more than 5 cm far from ESCC center were set as control group. 22 fresh and 175 paraffin-embeded ESCC specimens with corresponding adjacent tissues were randomly collected as study samples. With qRT-PCR, Western-blot and immunohistochemistry, the expression of Annexin A2 were detected at the mRNA and protein level. The correlation between Annexin A2 expression and clinicopathological parameters was analyzed. Results In 22 pairs of fresh ESCC and corresponding tumor adjacent tissues, the expression of Annexin A2 at mRNA level was significantly higher in tumor adjacent tissues (0. 06 ± 0. 06) than that in ESCC (0. 02 ±0. 02) (P＜0.05 ). Annexin A2 expression at protein level was also significantly higher in tumor adjacent tissues (0.95±0. 42) than ESCC (0.81±0. 36) (P＜0.05). In 175 paraffin-embeded ESCC specimens and corresponding adjacent tissues, the positive rate of Annexin A2 protein expression was 82. 3% (144/175) of the ESCC samples, which was lower than corresponding tumor adjacent tissues 92. 0% (161/175)(P＜0. 05). In addition, Annexin A2 expression was correlated with lymphoid node metastasis (P＜0.05) and pathological differentiation in patients with ESCC (P＜0.05). However, there was no apparent correlation with gross type (P＞0. 05). Conclusion The low expression of Annexin A2 in ESCC maybe played a potential role in the carcinogenesis, invasion and metastasis.