Carbonic anhydrase(CA) is one of a group of enzyme that contains zinc. They exist widely in the mammals. CA has at least 14 species of isoenzyme. Among them,CAIX is located at the downstream of VHL tumor suppressor gene,and it can be activated through HIF-1 pathway. CAIX primarily exists in the mucous membrane of gastrointestinal tract in normal human tissues and its expression is low. However,higher expression of CAIX can be detected in renal cell carcinoma,cervical carcinoma and lung cancer,it may be closely related with the occurrence and development of the tumor. In addition,we can predict the prognosis of tumor patients by analysis of the expression of CAIX,thus,it is considered that CAIX is a tumor specifi c antigen. Due to its highly specifi c expression in renal cell carcinoma,CAIX will become the potential target antigen of tumor vaccine as well as the target gene for the treatment of RCC. The application of CAIX shows high promise in RCC target therapy. Hence,it is defi nitely important for us to investigate the expression and regulation of the CAIX.

Objective To study the expression and enzyme activity of thioredoxin reductase 1 (TrxR1) in liver and peripheral blood of human and rats exposed to airborne arsenic through coal-burning as well as its role in liver injury of coal-burning-borne arsenic poisoning.Methods This study was divided into 2 parts.Part 1 was a population study:133 local residents exposed to airborne arsenic through coal-burning were selected as arsenic exposure groups including a non-patient group (25 cases),no obvious hepatopathy group (38 cases),mild (43 cases) and moderate to severe hepatopathy groups (27 cases) from areas affected by endemic arsenism in Guizhou Province.Thirty-four healthy residents from arsenic not affected areas were selected as controls.Peripheral blood samples were collected from all these people.The expression of TrxR1 mRNA was determined by real-time fluorescence quantitative PCR (qPCR),and enzyme activity of TrxR was tested by visible spectrophotometry.Part 2 was an animal experiment study:Thirty Wistar rats,weighing about 80-100 g,were divided into control group,drinking-waterborne arsenic poisoning group and coal-burning-borne arsenic poisoning group (including low,medium and high arsenic contaminated grain groups) by means of a table of random number according to body mass,6 rats in each group.The control group was fed with normal diet for 3 months; drinking-water-borne arsenic poisoning group and coal-burning-borne arsenic poisoning group were fed with 10 mg/kg As2O3 solution and different concentrations(25,50,100 mg/kg) of arsenic-containing feed,respectively,for 3 months.The expression of TrxR1 mRNA was determined by qPCR; protein expression level of TrxR1 in liver tissue was detected by immunohistochemistry,and enzyme activity of TrxR in serum and liver tissue was tested by visible spectrophotometry.Results The mRNA expressions of TrxR1 in peripheral blood were 1.599 8 (1.128 9-2.156 8),1.469 3 (1.146 1-1.976 3),1.203 6 (0.463 1-1.816 2) and 0.912 3(0.631 8-1.535 0),respectively,among non-patient group,no obvious hepatopathy group,mild and moderate to severe hepatopathy groups.Compared to the control group[1.649 7(1.161 1-2.380 2)],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The enzyme activity of TrxR in peripheral blood was (3.12 ± 0.76),(2.81 ± 0.84),(2.52 ± 0.73),(2.42 ± 0.76)U/ml,respectively,in those corresponding groups.Compared to the control group [(3.02 ± 0.70)U/ml],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The mRNA expressions of TrxR1 in peripheral blood were 1.05 ± 0.14,1.18 ± 0.18,1.04 ± 0.10 and 0.97 ± 0.13,respectively,among drinking-water-borne arsenic poisoning group,low,medium and high arsenic contaminated grain groups; all of which were lower than that in the control group (1.23 ± 0.15,all P ＜ 0.05) except that of the low arsenic contaminated grain group.The mRNA expressions of TrxR1 in liver tissue were 0.78± 0.10,0.83 ± 0.10,0.79 ± 0.09 and 0.77 ± 0.11,respectively; all of which were lower than that in the control group (0.94 ± 0.12,all P ＜ 0.05).The protein expression of TrxR1 in liver tissue was 310.33 ± 38.81,312.50 ± 23.36,305.67 ± 20.57 and 298.17 ± 23.52,respectively,among the arsenic poisoning groups; all of which were lower than that in the control group (348.50 ± 32.35,all P ＜ 0.05).The enzyme activity of TrxR in serum was (4.22 ± 0.73),(4.86 ± 0.63),(4.04 ± 0.57),(3.73 ± 0.64)U/ml,respectively; all of which were lower than that in the control group [(9.52 ± 1.08)U/ml,all P ＜ 0.05].The enzyme activity of TrxR in liver tissue was (14.82 ± 1.67),(18.76 ± 2.76),(14.90 ± 2.17),(11.55 ± 1.74) U/mg,respectively; all of which were lower than that in the control group [(23.71 ± 3.05)U/mg,all P ＜ 0.05].Conclusion Arsenic aggravates liver injury of coal-burning arsenic poisoning through down-regulating the expressions of TrxR1 mRNA and protein and reducing its enzyme activity as well.

Objective To observe the effect of electromyographic biofeedback combined with swallowing training on dysphagia after stroke.Methods Ninety-five patients with difficulty in swallowing after stroke were randomly divided into a treatment group (48 eases) and a control group (47 cases).The patients in the treatment group were provided electromyographic biofeedback and swallowing training; the patients in the control group received swallowing training only.The therapeutic effect was assessed with Kubota's drinking water test before treatment and 30d after treatment.Results Swallowing in both groups improved after treatment.The total recovery rate in the treatment group was 87.5％,and in the control group it was 68.0％,a significant difference.Conclusion Electromyographic biofeedback can improve the effectiveness of swallowing training after stroke.

Objective To observe the effect of electromyographic biofeedback therapy(EMGBFT)on dysphagia in stroke patients.Methods Fifty-three stroke patients with dysphagia were divided randomly into an EMGBFT group and a control group.The patients of EMGBFT group were given EMGBFT,electrical stimulation therapy (EST)and dysphagia training,while those in the control group were given EST and dysphagia training.All the patients were assessed with Kubota drinking test before treatment and 30 days after treatment.Results After treatment swallowing function of patients in both groups improved(P ＜0.05).The effective rate was 76.92％ in EMGBFT group and 55.56％ in control group,with statistically significant difference between the two groups(P ＜ 0.05).It showed that the EMGBFT group has significantly better outcome than the control group after treatment(P ＜ 0.05).Conclusions EMGBFT combined with regular rehabilitation therapy can improve patient's motor and swallowing function.

Objective To explore family functioning and related factors in anxiety disorder(AD)adoles cents.Methods 84 adolescent patients with AD and 124 controls without mental disorder matched by gender,age,and residential township were assessed with Family Adaptability and Cohesion Scale(FACES-Ⅱ),Family Assessment Device.Chinese version(FAD-CV)and demographic questionnaires.Results There were statistical differences between case and control groups in family economic conditions(P<0.05).There was lower family cohesion in AD group(64.82±9.63)than that in the control group(69.72±8.91);the scores of communication,emotional involvement and behavioral control were higher in AD group((2.34±0.45),(2.52±0.44),(2.46±0.32))than that in controls((2.12±0.34),(2.36±0.42),(2.24±0.24)),all P<0.05.Logistic regression analysis showed that better family economic condition(OR=0.784;95%CI:1.459-3.396)and higher cohesion of family(OR=0.969;95%CI:0.942～0.998)were protective factors for AD,while over-strict behavioral control (OR=2.181;95%CI:0.993～2.441)was a risk factor for AD.Conclusion There are different family functioning characters between AD and controls,and the intimacy relationship and suitable behavior control in family may improve the mental health of adolescents.

Objective To observe the effects of pulsed electromagnetic fields (PEMFs) on behavior and mechanical paw withdrawal thresholds (MPWTs) after acute skeletal muscle contusion (ASMC) in rats, and to in-vestigate the application of PEMFs in rats with ASMC during the early stage. Methods Forty-two Sprague-Dawley rats were randomly divided into a PEMF group (P group) , control group (C group) and blank control group ( BC group). ASMC models were set up in groups P and C, and no intervention was applied in the BC group. A PEMF was administered to animals in the P group immediately after the ASMC was inflicted. The behavior of the rats in each group was then observed. The MPWT of each rat was tested 2 days before and 0, 12, and 18 hours after the ASMC was inflicted). Results In the P and C groups, MPWT of the left hind paw at the 12th and 18th hour after ASMC was significantly lower than the baseline pain threshold 2 days before the ASMC. At 18 hours, the MPWT was signifi-cantly higher than at 12 hours in the P group. MPWT at 12 hours in the P group and at both 12 and 18 hours in the C group were significantly lower than in the BC group. MPWT in the P group at 18 hours was significantly higher than in the C group. Conclusions The behavior of rats treated with PEMF immediately after ASMC was improved, and their pain threshold was still elevated 18 hours later.

Objective To observe the effects of pulsed electromagnetic fields (PEMFs) on histological changes and myogenic differentiation factor D (MyoD) expression in rats with acute skeletal muscle contusion ( ASMC), and to explore the effects of PEMF therapy on rats with ASMC in its very early stages. Methods Forty-two rats were randomly divided into three groups : a treatment group, a control group and a blank control group. ASMC models were established with all the animals in the treatment and control groups. PEMF treatment was admin-istered to the treatment group immediately after the establishment of the ASMC model. Seven rats in each group were sacrificed at the 12th and 18th h after the models were set up. Their triceps surae muscles were sampled and treated with haematoxylin-eosin staining for study using immunofluorescence techniques and a fluorescence microscope. Re-suits In the control group at the 12th h and 18th h, HE staining showed pale cytoplasm and polymorphism in the cell nuclei ; in the treatment group these effects were significantly lighter, but in both groups it was more serious than in the blank control group. In the treatment and control groups, the fluorescence intensity of MyoD at the 18th h was higher than at the 12th h, and at each time point in both groups it was higher than in the blank control group. At the 18th h, fluorescence in the treatment group was stronger than in the control group. Conclusion MyoD expression in rats with ASMC is upregulated by thel8th h after early PEMF treatment. This might be one of the mechanisms ac-celerating the regeneration of skeletal muscles after trauma.

BACKGROUND:The metastatic potential of hepatocelular carcinoma cels is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cels on metastasis of hepatocelular carcinoma is of great significance for improving the lifetime of hepatocelular carcinoma patients. OBJECTIVE:To explore the biological effect of human bone marrow mesenchymal stem cels on hepatocelular carcinoma cels with different metastatic potentials. METHODS:Human bone marrow mesenchymal stem cels and hepatocelular carcinoma cel suspension with high and low metastatic potentials were respectively injected into the Transwel chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as wel as cel counting method was used to observe the changes in the invasion ability of hepatocelular carcinoma cels. The effects of human bone marrow mesenchymal stem cels on the proliferation of hepatocelular carcinoma cel suspension with high and low metastatic potentials were determined using cel counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cel death protein 5. RESULTS AND CONCLUSION:(1) The number of migrated hepatocelular carcinoma cels was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P> 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group (P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P< 0.05), but the expression of programmed cel death protein 5 showed no changes (P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cel death protein 5 was both increased dramaticaly (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cels reduce the invasion ability of hepatocelular carcinoma cels, and enhance their ability of proliferation.

Background and purpose: The increased of specific expression of prostate cancer antigen 3 (PCA3), as one of long non-coding RNA, has been observed in prostate cancer, indicating that PCA3 may contribute to the development of prostate cancer. To further study its roles in prostate cancer, we construct a lentivirus expression vector carrying the whole PCA3. Methods: PCA3 was amplified from prostate cancer cell line LNCaP by reverse transcriptase polymerase chain reaction (RT-PCR). After the sequence was proved to be correct, we recombined the pCDH-CMV-MCS-EF1-copGFP-PCA3. After transformation into E.coli cells, the candidate clones were identified by PCR amplifying, restricting enzyme digestion analysis and DNA sequencing, and the viral titer was determined. Results: Through the Blast analysis software, we compared the results of PCA3 sequence amplified by PCR with GeneBank sequence, ifnding that the homology is 99.8%. The lentivirus vector was constructed successfully, and the virus in the supernatant reached a titer of 2*108. Conclusion: The successful construction of the lentivirus vector encoding PCA3 not only lays the foundation for the further research into the effect of PCA3 gene on the prostate cancer but also provides a new therapy for advanced prostate cancer.