Background The pathogenesis of pterygium has been in the study.Relative molecular biology study showed that pterygium is a tumor-like lesion,and based on overseas literatures,human papillomavirus (HPV) is positively expressed in 100％ patients with pterygium in some region.However,if this result is suitable for Chinese patients is unclear.Objective This study was to identify the role of human HPV in the pathogenesis of pterygia in Wuxi area.Methods Forty-eight pterygium specimens including 7 recurrent pterygia and 41 primary pterygia were collected during the operation,and these patients were from Wuxi area.Two cervical carcinoma specimens and 2 conjunctiva specimens from normal donors were obtained as positive control and negative control respectively.The fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect HPV DNA of specimens.Results The amplified curves of HPV 6,11 of pterygium specimens,cervical carcinoma specimens and normal specimens were all below the positive quality control curve,but the amplified curves of HPV16,18 were above the quality control curve in cervical carcinoma specimens; while those of pterygium specimens and normal conjunctival specimens were all below the quality control curve.HPV16/18 was identified in 2 cervical carcinoma specimens,but no HPV6/11 was detected in 2 cervical carcinoma specimens.However,HPV DNA expression in primary and recurrent pterygias were absent.Conclusions According to these results,HPV is not the primary cause for the pathogenesis of pterygium in Wuxi region.

Objective To evaluate the effect of therapeutic touch on continuous positive airway pressure titration with obstructive sleep apnea hypopnea syndrome (OSAHS) patients.Methods Fortyeight patients diagnosed with OSAHS were randomly allocated to the therapeutic group and the control group with 24 patients in each group.Therapeutic touch was used in the therapeutic group before CPAP titration and the control group received usual care.Anxiety was measured post intervention with State-trait Anxiety Inventory(SAI) and Visual Analogue Scale(VAS).CPAP therapy satisfaction was also evaluated by VAS.Results Compared with the control group,those patients with therapeutic touch showed significant reductions in SAI scores,better sleep quality,and higher satisfaction with CPAP treatment.Conclusions Results suggest that 15-min of therapeutic touch pre CPAP titration night significantly reduced anxiety and improved sleep quality in patients with OSAHS.

Objective:To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene,and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumor-bearing mice.Methods:The specific siRNA of Survivin gene was designed and synthesized,and a recombinant adenovirus AdEGFP-siRNA was subsequently constructed.SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups:siRNA+radiotherapy and siRNA groups(intratumoral injection of AdEGFP-siRNA),siRNA(-)group(injected every other day with AdEGFP-siRNA[-],2?108 pfu/100 ?l per time,total 5 times),pure radiotherapy group and blank control groups(injected with the same volume of normal saline).On day 10,12,14,and 16,the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy.The tumor volumes were measured regularly.The expression of Survivin in tumor tissues was determined immunohistochemically.Results:Adenovirus AdEGFP-siRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene(EGFP)was successfully recombined.The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFP-siRNA,with the inhibition rate being 56.2%.The inhibition rate in AdEGFP-siRNA therapy + radiotherapy increased to 82.6%.Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells.Conclusion:The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer;meanwhile,it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.

Objective To investigate the clinical features of and gene mutations in a Chinese Han pedigree with piebaldism. Methods Clinical data were collected with informed consent from a pedigree with piebaldism, processed and documented. A clinical genetic analysis was conducted and pedigree chart was drawn. Genomic DNA was extracted from the peripheral blood of 14 patients and 40 unaffected individuals in the family as well as 50 unrelated human controls, and subjected to the amplification of 21 exons and flanking sequences of the KIT gene by PCR. Sequence analysis was performed by Mutation SurveyorTM. Results There were 73 members in the family, and of them, 14 were diagnosed with piebaldism according to typical clinical features. Piebaldism was inherited in an autosomal dominant pattern in this family. A heterozygous 4-base insertion mutation 1900insATGA in exon 13 of KIT gene was identified in all the 14 affected family members, which resulted in a frame-shift mutation at codon 634 and produced a premature translation termination codon. This mutation was undetected in either the unaffected family members or unrelated controls. Up to the time of this writing, this mutation had not been previously reported. Conclusion The novel mutation 1900insATGA in the KIT gene may be the cause of clinical phenotype of piebaldism in the family.

Objective To investigate the effects of ultrasound irradiating hydroxycamptothecin (HCPT)-loaded microbubbles on apoptosis in human hepatoma SMMC-7721 cells, and to clarify its probable mechanism. Methods The SMMC-7721 cells were randomly divided into six groups as follows: control group, blank liposome microbubbles plus ultrasound, HCPT-loaded microbubbles, HCPT, HCPT plus ultrasound,and HCPT-loaded microbubbles plus ultrasound. Apoptosis was ascertained by Annexin V-Fire/ PI and TUNEL methods. The expression of Bcl-2 and Bax protein was detected by immunocytochemistry staining technique. Results In the group of HCPT-loaded microbubbles plus ultrasound,the apoptosis rate at early term was found to reach (12.18 ± 1.38)% by Annexin V-Fitc/PI assay, brown-colored positive apoptotic cells were observed and the apoptosis rate at advanced term was found to reach (34.25 ± 1.83)% with TUNEL method. The expression of Bcl-2 was found to decrease,the expression of Bax increase,and the ratio of Bcl-2/bax significantly decrease by immunocytochemistry staining technique in the same group. Differences with other groups were significant. Conclusions The method of ultrasound irradiating HCPT-loaded microbubbles can notability induce human hepatoma SMMC-7721 cells apoptosis. The decrease in the ratio of Bcl-2 to Bax might be responsible for cells apoptosis.

To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

OBJECTIVETo observe the effect of Polygala tenuifolia Willd YZ-50 on the mRNA expression of brain-derived neurotrophic factor (BNDF) and its receptor TrkB in rats with chronic stress depression.es.

METHODSNormal male Wistar rats were divided in to control group, model group, desipramine (20 mg/kg) group, and low and high-dose (2.8 and 5.6 g/kg) YZ-50 groups. The total RNA was extracted from the rats with chronic stress depression, and the mRNA expression of BDNF and TrkB was detected by RT-PCR.

RESULTSCompared with the model group, YZ-50 at both low and high doses significantly increased the mRNA expression of BDNF and TrkB in the hippocampus of rats with chronic stress depression, and the effect was more obvious in the high-dose group (P<0.01).

CONCLUSIONYZ-50 can up-regulate the expression of BDNF and TrkB mRNA to promote the recovery of the neurons from chronic stress-induced damages and produces anti-depressant effect.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Depression ; drug therapy ; etiology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Polygala ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Receptor, trkB ; genetics ; metabolism ; Stress, Physiological ; physiology ; Up-Regulation ; drug effects

Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj（o）gren syndrome (SS) and 20 cases with non-Sj（o）gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.

BACKGROUNDKAI1 is a new identified metastasis-suppressor gene whose expression in many types of tumors has been reported. The aim of study is to investigate the role of KAI1 protein in development of lung cancer and its values in predicting the prognosis of lung cancer.

METHODSThe expressions of KAI1 protein were detected in benign pulmonary disease tissue, precancerous disease tissue, lung cancer tissue and metastatic lung cancer tissue in local lymph node using tissue microarray and immunohistochemical method. The relationship between expression of KAI1 protein and clinicopathological parameters of patients with lung cancer was analyzed by Chi-Square test and Fisher exact test.

RESULTSThe positive rate of KAI1 expression was 100.0% in 10 cases of benign pulmonary diseases, 66.7% in 12 cases of precancerous diseases, 24.7% in 89 cases of primary lung cancer and 0 in metastatic lung cancer tissue in local lymph node respectively. The KAI1 protein expression in primary lung cancer tissues had no remarkable relationship with age and gender of the patients and the location of cancer, but had significant relationship with the histological type and differentiated degree of tumor, P-TNM stages and lymph node metastatic status.

CONCLUSIONSThe abnormal expression of KAI1 protein may participate in malignant progression of lung cancer. Its downregulation may promote the invasion and metastasis of tumor cell. Detection of the expression of KAI1 protein may be helpful to predict the prognosis of lung cancer.