BACKGROUND:The dynamic changes of hypoxia inducible factor-1 alpha (HIF-α) and inducible nitric oxide synthase (iNOS) genes in the pulmonary artery wall during the process of hypoxic pulmonary hypertension (HPH) development need to be investigated.OBJECTIVE: This study was to observe the gene expressions of HIF-α and iNOS in the pulmonary artery wall at the different hypoxic time points, and to investigate their effects in HPH development.DESIGN: Controlled observation animal experiment.SETTING: Department of Respiration, Second Hospital Affiliated to Nanhua University.MATERIALS: Forty healthy male Wistar rats of clean grade, aged 6-8 weeks, with body mass of (220±10) g were involved in this study. They were randomized into control group (n =8) and hypoxia group (n =32). Four time points, i.e.3, 7, 14, and 21 days after hypoxia were set for the animals in the hypoxia group, 8 rats at each time point.METHODS: This study was carried out in the Institute of Oncology, Nanhua University between August 2004 and December 2005. Rats in the hypoixa group were treated according to the method reported by Li et al. Hypoxia treatment was omitted for the rats in the control group. At each time point, the rats were anesthetized, and then a micro-catheter was inserted into the right jugular vein and connected to a multichannel physiologic recorder, which was used for detecting the mean pulmonary arterial pressure (mPAP). The heart of each euthanized rat was taken out, and its right ventricle (RV), and left ventricle and septum (LV+S) were weighted. Right ventrical hypertrophy index (RVHI) reflected right ventricle hypertrophy degree. Right upper lung tissue of rat was harvested for haematoxylin & eosin (HE) staining and elastic fiber staining. Pathological image analysis software was used to determine pulmonary arterial wall area/total vascular area, lumina area/total vascular area, smooth muscle cell density in the media of pulmonary arteriole, and media thickness of pulmonary arteriole, which were used as remodeling indexes of pulmonary arteriole. HIF-1α and iNOS in the pulmonary arteriole were performed in situ hybridization and immunohistochemical detection. The mean absorbance of pulmonary arteriole wall was used as the relative content of mRNA expression and protein level.MAIN OUTCOME MEASURES: ①Changes in mPAP, RV hypertrophy degree and remodeling indexes of pulmonary arteriole of rats. ② Expressions of HIF-α and iNOS as well as their correlations with mPAP and remodeling indexes of pulmonary arteriole.RESULTS: All the 40 Wistar rats were involved in the final analysis. ①At hypoxia 7 days, mPAP was significantly higher than that of control group (P ＜ 0.05). mPAP reached to the high level at hypoxia 14 days, and then maintained at this level. ② At hypoxia 14 days, RVHI was higher than that of control group (P ＜ 0.05). ③ At hypoxia 7 days, pulmonary arteriole wall was thickened, and lumina became narrowed. There were significant differences in pulmonary arterial wall area/total vascular area, lumina area/total vascular area between hypoxia group and control group (P ＜ 0.05). At hypoxia 14 days, smooth muscle cell density in the intima-media of pulmonary arteriole, and intima-media thickness of pulmonary arteriole were significantly increased (P ＜ 0.05). At hypoxia 21 days, lumina was further narrowed, and obvious smooth muscle hyperplasy was found. HIF-1α and iNOS mRNA expression presented weak-positive in the control group; The relative content of HIF-1α mRNA did not significantly alter at hypoxia 3 and 7 days, but was obviously increased at hypoxia 14 days ,and after this, it maintained at high level. iNOS mRNA was markedly higher than that in the control group at hypoxia 3 days, reached its peak at hypoxia 7 days, was close to the level in the control group at hypoxia 14 days, and enhanced again at hypoxia 21 days, but it was still lower than that at hypoxia 3 days. ⑤HIF-α was mainly found in the intima and media, while iNOS was found in the whole layer of vessels. iNOS was weakly expressed in the intima and media of pulmonary vessels in the control group. No obvious difference in iNOS existed at hypoxia 3 days as compared with control group. iNOS was obviously expressed in media and intima at hypoxia 7 days. Vascular media was thickened at hypoxia 14 days, and the expression of iNOS was enhanced. iNOS was not found in the vascular adventitia in the control group, but was found in the vascular adventitia in the hypoxia group. ⑥mPAP was positively correlated with pulmonary vascular remodeling (r =0.976, P ＜ 0.01 ), and HIF-1α mRNA was positively correlated with iNOS protein(r =0.927, P ＜ 0.05).CONCLUSION: Both HIF-1α and iNOS exert effects in the process of HPH development of rats, and HIF-1α and iNOS gene expression may be mutually regulated.

The age of 18 is an important criterion in judicial trial, immigrant and competitive sports. Consequentially, the estimation of age 18 is a key issue in forensic practice and research. The extremitas sternalis claviculae, iliac crest, third molar, and the proximal limb of the limb bone were usually used as indictors of age 18. The results of the previous studies demonstrated that those indictors could be beneficial to the estimation of age 18. The X-ray, CT, ultrasound and MRI of different indictors were widely utilized for the estimation of age 18, particularly the thin-layer CT. But due to the non-radiation, MRI will be a trend for forensic age estimation in the future. Whilst in the previous studies, the descriptive analysis was applied for the estimation of age 18, but due to the low statistic efficiency, it is unsuitable for forensic age estimation, and the future studies should pay attention to the high efficiency statistical methods, for instance, the ROC curve or the data mining.

Objective To study the effects of percutaneous liver tumor injection combined with the clinical efficacy of transcatheter hepatic artery chemoembolization in the treatment of advanced liver cancer,and to provide ref-erence for clinical treatment.Methods 22 patients using percutaneous liver tumor injection combined with transcath-eter arterial chemoembolization for treatment were selected,with which 1 month follow-up after discharge.Situation of patients with percutaneous liver tumor injection and transcatheter hepatic artery chemoembolization was analyzed,and the changes of the patients in the following -up of survival time,tumor volume and clinical symptoms were also ana-lyzed.Results Among the patients of postoperative recheck after 6 weeks,6 cases were complete remission,there were partial remission in 8 cases,6 cases of stable,2 cases of progress.Follow up to 2013 December,the patients'sur-vival time was 17-82 months,the average survival time was (55.71 ±13.47)months.After treatment,4 cases of patients'tumor diameter reduced 1 -3cm,18 cases of tumor diameter reduced 3 -5cm,19 cases of liver area pain symptoms for more than half a year of remission,3 cases of liver area pain relief time less than half a year.During the follow -up period,12 patients died of multiple organ failure.Conclusion The development of percutaneous liver tumor injection combined with transcatheter hepatic arterial chemoembolization therapy can delay the development of the disease in patients with advanced HCC,and prolong the survival time.

Objective To analyze causative factors for and therapeutic effect of total glucosides of paeony (TGP) on palmoplantar pustulosis (PPP).Methods Ninety patients with PPP were recruited in this study.A questionnaire survey was carried out to collect clinical data from these patients,and a patch test to identify contact allergens for these patients.All the patients were randomly divided into two groups:observation group treated with TGP capsule 0.6 g thrice a day,and control group treated with tripterygium glycosides tablets 20 mg thrice a day.Both groups topically applied mometasone furoate cream once a day.The efficacy and side effects were evaluated 4 and 8 weeks after initiation of treatment.Results Of the 90 patients,the ratio of male to female was 1 ∶ 1.64,smokers amounted to 42.22％,and 23 (26.67％) reported a history of infection before the onset of PPP.As the patch test showed,potassium dichromate and nickel sulfate were the most common contact allergens in these patients.The response rate was significantly higher in the observation group than in the control group after 4 weeks of treatment (64.44％ vs.40.00％,x2 =5.388,P ＜ 0.05),but similar between the two groups after 8 weeks of treatment (88.89％ vs.80.00％,x2 =1.353,P ＞ 0.05).No significant difference was observed in the occurrence of adverse reactions between the two groups (17.78％ vs.11.11％,x2 =0.809,P ＞ 0.05).Conclusions Smoking may play a certain role in the development of PPP,and infection seems to be a primary inducing factor.TGP combined with mometasone furoate cream is effective for the treatment of PPP.

Objective To investigate the antioxidative effects of all-trans retinoic acid (ATRA) on the intestinal ischemia reperfusion.Methods Thirty-two male SD rats were randomly divided into the sham operation group,occlusion group,dimethyl sulfoxide (DMSO) group and the ATRA group according to the random number table.There were 8 rats in each group.Rat models of intestinal ischemia reperfusion were established by clamping the superior mesenteric arteries for 60 minutes,and then restore the blood flow for 120 minutes.Superior mesenteric arteries were only separated without clamping in the sham operation group.Rats in the ATRA group received ATRA pretreatment through intragastric infusion at the dosage of 15 μg/g for 5 days,and then ATRA pretreatment at 6 hours before operation.Rats in the DMSO group received intragastric infusion of DMSO at the same dosage.The concentration of ATRA at 5 hours before operation was detected by high performance liquid chromatography.The pathomorphological changes of the ileal mucosa were detected by hematoxylin-eosin staining,and the Chiu's scores on the ileal mucosa were evaluated.The serum content of diamine oxidase (DAO),tissue level of malonaldehyde (MDA) and activity of superoxide dismutase (SOD) were detected by colorimetry.Protein expression of manganese superoxide dismutase (MnSOD) in the ileal tissues was detected by Western blot.All data were analyzed using the analysis of variance or LSD-t test.Results The concentrations of ATRA in the ATRA group was (827 ±276) μg/L,which was significantly higher than (48 ± 12) μg/L of the sham operation group,(55 ± 15) μg/L of the occlusion group and (63 ± 20) μg/L of the DMSO group (t =11.242,11.138,11.013,P ＜ 0.05).The morphology of the ileal mucosa was normal in the sham operation group,while the ileal mucosa was severely damaged in the occlusion group and the DMSO group.The injury of the ileal mucosa in the ATRA group was slight.The Chiu's scores of the occlusion group,DMSO group and the ATRA group were 3.83 ±0.77,3.92 ± 0.87 and 2.42 ± 0.75,which were significantly higher than 0.37 ± 0.28 of the sham operation group (t =9.803,10.040,5.793,P ＜0.05).The Chiu's scores of the ATRA group was significantly lower than that of the occlusion group and the DMSO group (t =4.009,4.247,P ＜ 0.05).The DAO levels of the occlusion group,DMSO group and the ATRA group were (26.3 ±4.4)U/L,(25.1 ± 4.3)U/L and (20.8 ±3.8)U/L,which were significantly higher than (14.2 ± 1.9) U/L of the sham operation group (t =6.493,5.835,3.534,P ＜ 0.05).The level of DAO of the ATRA group was significantly lower than that of the occlusion group and the DMSO group (t =2.959,2.301,P ＜0.05).The levels of MDA of the occlusion group,DMSO group and the ATRA group were (16.9 ± 4.0) μmol/g,(16.0 ± 3.5) μmol/g and (11.3 ± 3.1) μmol/g,which were significantly higher than (5.4 ± 1.0) μmol/g of the sham operation group (t =7.397,6.821,3.821,P ＜ 0.05).The level of MDA of the ATRA group was signifcantly lower than that of the occlusion group and the DMSO group (t =3.575,3.000,P ＜ 0.05).The SOD activity of the occlusion group,DMSO group and the ATRA group were (108 ±22) U/mg,(98 ± 19) U/mg and (181 ± 38)U/mg,which were significantly lower than (243 ± 37)U/mg of the sham operation group (t =8.939,9.647,4.106,P ＜ 0.05).The SOD activity of the ATRA group was significantly higher than that of the occlusion group and the DMSO group (t =4.833,5.541,P ＜ 0.05).The relative protein expressions of the MnSOD of the occlusion group and the DMSO group were 0.36 ± 0.08 and 0.28 ± 0.07,which were significantly lower than 0.93 ± 0.13 of the sham operation group (t =8.972,10.101,P ＜ 0.05).The relative protein expression of the MnSOD of the ATRA group was 0.80 ± 0.19,which was significantly higher than that of the occlusion group and the DMSO group (t =6.948,8.077,P ＜ 0.05),while it was not significantly different from that of the sham operation group (t =2.024,P ＞ 0.05).Conclusion Through up-regulating the expression of MnSOD and improving the antioxidative capacity of tissue,ATRA pretreatment can attenuate intestinal ischemia and reperfusion injury.

Objective To explore the sensitivity and specificity of conventional eytogeneties(CC),nested-reverse transcriptase polymerase chain reaction(nested-RT-PCR) and dual-color and dual-fusion fluorescence in situ hybridization(D-FISH) technique in monitoring the tumor load of chronic myeloid leukemia (CML) during treatment.Methods CC,nested-RT-PCR and interphase D-FISH were simultaneously carried out to detect the tumor load of 5 CML patients during treatment with interferon-Mpha(IFN-α).Results 24 specimens from 5 CML patients before and after IFN-α therapy were investigated and the results showed that 23 specimens were Ph+ with different positive ratios by CC.All specimens were bcl-abl mRNA (+) by RTPCR.The Ph-ber-abl+ specimen from case 2 after 75 months of post-treatment showed 4.5%bcr-abl+ cells by FISH.2 specimens from case 1 at 22 mortths,26 mortths of post-treatment.2 specimens from cage 5 after 12 months,16 menths of post-treatment and 2 specimens from case 4 after 6 menths,10 menths post-treatment with same Ph+ ratio respectively were investigated by FISH and showed 47.5% and 39.5%,74.0%and 60.5%,99.0% and 99.5% bcr-abl+ cells,respectively.Conclusion CC can be used as a basic tool to monitor the change of tumor load in CML during treatment. When CC can't evaluate precisely dynamic changes of tumor load and when tumor load in patient with treatment were too low to detect Ph bv CC while bcr-abl mRNA was still positive by RT-PCR,FISH Call be used to detect precisely tumor load and monitor dynamic change of the disease.More sensitive RT-PCR was used to monitor tumor load when it was negative to bcr-abl by FISH during treatment.

Objective To establish an otitis media with effusion model in rat,and provide a pure physical approach of animal model for researching the otitis media with effusion.Methods Twenty-four clean grade male SD rats were selected.Only the right ears were used for the model group,the contralateral ears were used as the control group.The animal model of otitis media with effusion was established by blocking eustachian tube with cork stopper through the right side mandible,the hearing was evaluated by ABR and tympanogram configurations,the pathologic changes of middle ear and eustachian tube mucous membrane were examined by light microscopy.Results Twenty out of 24 ears of rats' of hearing threshold increased from 34.25?5.45 dB to 57.63?5.46 dB (P

ObjectiveTo evaluate the prevalence of occult hepatitis B infection in blood donors and to explain the possible molecular mechanfism of occult hepatitis B infection.Methods Enzyme linked immunosorbent assay (ELISA) method was used for detection of hepatitis B virus (HBV)markers in serum samples of 594 donors which were collected from blood bank with HBsAg negative results.Nested-polymerase chain reaction (PCR) was performed to detect serum HBV DNA.In donors with occult HBV infection,the serum HBV markers were quantitatively detected by Abbott nested-PCR kit.The PCR products of S region were sequenced and sequence alignment was performed to analyze relevant virus mutations.Eleven HBsAg positive patients were randomly recruited as positive controls and S region was amplified and sequenced.The difference of S region sequences was compared between patients with occult HBV infection and HBsAg positive HBV infection.ResultsAmong 594 HBsAg negative donors,15 were diagnosed with occult HBV infection with the incidence of 2.5 ％.No correlation was found between results of serum HBV markers and occult HBV infection.Sequencing results of HBV S region were obtained from 10 cases,which revealed mutations of HBV.The amino acid mutations in the “a” determinant cluster were found in three patients,which were I126T,T140I and T140I,respectively.On the contrast,mutation in the “a” determinant cluster of T131 N was only found in one positive control.ConclusionsThe occult HBV infection exists in blood donors with negative results for HBsAg test.Genetic mutation may play a role in the occult HBV infection.

Objective Analysis the risk factors which maybe caused the cerebral hemorrhage after the liver transplantation, and then make out the counter nursing measures. Methods Selected 12 patients with cerebral hemorrhage after the liver transplantation as the experimental group, and then matching 48 patients without cerebral hemorrhage after the liver transplantation as the control group. Analysis the factor which related the cerebral hemorrhage. Results The factors which related the cerebral hemorrhage after the liver transplantation included in the Intensive Care Unite, heme level, Child level, APTT, albumin, blooding volume during the operation, operation time and the postoperative blood platelet level. The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage. Conclusion The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage, we should carry out counter nursing measures to prevent them.

Iliac arteriovenous ifstula (AVF) usually manifests in a wide range of symptoms similar to typical deep venous thrombosis (DVT), which often lead to delayed diagnosis or misdiagnosis. We reported a 51-year old woman who was performed lumbar discectomy and showed a progressive abdominal distention, dyspnea, and swollen left leg. She was initially diagnosed as deep vein thrombosis and the ifnal diagnosis was arteriovenous ifstula. hTe ifstula was successfully sealed by an endovascular covered stent. No further recurrence was found atfer a half year’s follow-up. hTis article summarized the experience regarding iliac arteriovenous ifstula misdiagnosed, and discussed the differential diagnosis between arteriovenous ifstula and pulmonary thromboembolism causedby deep vein thrombosis.