Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250－700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.

Background Corneal blindness is one of the major blinding eye diseases in China.With the development and progress of tissue engineering technology,tissue-engineered corneas offers a new approach to the treatment of corneal diseases.To select and cultivate ideal seed cells is a foundation of construction of tissueengineered corneas.Objective This study was to evaluate the efficiency of stripe off the Descemet membrane with endothelium plus enzymic digestion in the isolation of corneal endothelial cells and analyze the bionomics of rabbit corneal endothelial cells (CECs) in vitro.Methods Descemet membrane was stripped from fresh cornea of New Zealand rabbit under the dissection microscope.Descemet membrane with endothelium was incubated in trypsin and EDTA solution at 37 ℃ and then purified for CECs subculture in vitro.The morphology of the cultured cells was observed under the inverted microscope and marked by CM-Dil dye solution.Then the shape of the cells was observed by hematoxylin and eosin staining and the cells were identified for the expression of neuron specific enolase (NSE) using immunochemistry.The viability of the cells were evaluated by trypan blue staining.The surface structure of the cells were examined under the scanning electron microscope.Intercellular zonula occludens-1 (ZO-1) was identified by immunofluorecsence staining.Results A large number of purified CECs were obtained from Descemet membrane with endothelium through enzymic digestion.Cultured cells grew well and formed monolayer 5-7 days later with the cobblestone stone-like arrangement.The survival rate of the cells was 95％.CECs presented with the red annular fluorescence for CM-Dil with the labeling rate ＞90％.NSE was positively expressed in the cytoplasm.Polygon CECs were seen by hematoxylin and eosin staining and showed the brown staining.Abundant microvilli on the cellular surface and interconnected foot process were seen under the scanning electron microscope.ZO-1 showed the green fluorescence.Conclusions The method of striping off the corneal Descemet membrane with endothelium plus enzymic digestion can obtain abundant CECs.Cultured cells have good biological properties.This study may offer a feasible application in the engineering of corneal transplant membrane.

Adult ; Atrioventricular Block ; complications ; Cardiomyopathies ; complications ; diagnosis ; Humans ; Male ; Sarcoidosis ; complications ; diagnosis

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

Purpose To investigate the influence on shear wave velocity (SWV) by different probe frequency, scanline angle and ROI depth. Materials and Methods The elasticity QA Phantoms was used in the study. First the SWV was measured by curve probe at depths of 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm and 8 cm. The scanline angle changed from 0°from midline, 1°-15°, 16°-30°and 31°-45°. Then the SWV was measured again by using line probe at depths of 1 cm, 2 cm, 3 cm and 4 cm. The measured data were analyzed statistically. Results The SWV showed difference by using the two probes (P<0.05). As depth increased, the measurement by curve probe changed from overestimate to underestimate, while the measurement by line probe always showed underestimate from standard value. When the curve probe was used, the angle and depth affected SWV interactively (F=85.87 and 35.96, P<0.01), and had interactive effects (F=17.63, P<0.01). When the scanline angle was within 15°, the angle had little inlfuence upon the measurement. When the scanline angle was 0 °, the measurement was the closest to the standard value at the depth of 4 cm, and the maximum deviation occurred at the depth of 5 cm. The inlfuence of depth upon measurement had signiifcant difference when the line probe was used (P<0.05). The measurement was the closest to the standard value at the depth of 1 cm, and the maximum deviation occurred at the depth of 4 cm. Conclusion Factors like probe frequency, scanline angle and ROI depth can affect the SWV. When the curve probe is used, the accurate measurement occurs at the depth of 4 cm when the scanline angle is within 15°. When the line probe is used, the accurate measurement occurs at the depth of 1 cm.

0.05).Seroquel showed high incidence rate of drowsiness,dizziness and weight gain(26.1%,23.9%,17.3%),chlorpromazine showed high incidence rate of extrapyramidal side effect,pyknocardia and erect collapse(60.9%,39.1%,32.6%).CONCLUSIONS:There was no significant difference of curative effect for the treat-ment of schizophrenia between seroquel-treated group and chlorpromazine-treated group,but seroque showed low side effect and high safety.

Background Interleukin (IL)-1a-889C/T,IL-1β-S11C/T,+ 3962C/T and IL-1Ra-2 are different morphology of single nucleotide of interleuki-1 (IL-1).The potential relationship of IL-1 polymorphisms with Beh(c)et disease has been studied in several populations and groups.However,study outcomes are controversial for a long duration.Objective This Meta-analysis was to determine whether the IL-1 gene polymorphisms is associated with the pathogenesis and development of Beh(c)et disease.Methods The literature of the assoiation between IL-1 polymorphisms and Beh(c)et disease was retrieved from Medline,EMBASE,Cochrane Library,Web of knowledge,Google Scholar,Wanfang and CNKI databases.All the published original articles (case-control studies) were reviewed with the deadline May 31,2013,and the languages were limited to English and Chinese.Included reports were evaluated based on Newcastle-Ottawa Scale (NOS) score.The potential influence of IL-1α-889TT gene,IL-1β-3962C,IL-1β-511T and IL-1Ra-2 polymorphisms on Beh(c)et disease were analyzed.RevMan5.0,the Cochrane collaboration software program,was used to prepare and complete this review.The effect size was assessed using fixed effect model in the index with lower heterogeneity (I2 ＜50％) or randomized effect model in the index with higher heterogeneity (I2＞50％).Results The literature search resulted a total of 370 cases-controlled studies,and 7 studies met the included criteria,with the NOS scores ≥ 8.The total patients were 499 and normal controls were 708.Meta-analysis was performed on several populations.Overall,the polymorphisms of IL-1β-3962C allele increased the susciptibility of Beh(c)et disease (OR =1.41,95％ CI:1.06-1.88,P =0.02),and IL-1α-889TT genotype reduced the risk of Beh(c)et disease (OR =0.61,95 ％ CI:0.40-0.92,P =0.02).However,There was no significant association between variants of IL-1β-511T allele (OR =0.84,95％ CI:0.58-1.23,P=0.38) or IL-1RA-2 allele (OR =1.25,95％ CI:0.50-3.14,P=0.63) and Beh(c)et disease susciptibility.Conclusions These results suggest that Beh(c)et disease is associated with the IL-1 gene polymorphisms at the locations α-889C/T and β-3962C/T.A larger sample size clinical data still are need to confirm this conclusion.

ObjectiveTo evaluate the potential value of acoustic radiation force impulse (ARFI)elastography in the characterization of solid liver tumors.MethodsForty-three patients with 56 liver tumors were evaluated with ARFI,which included 21 patients with hepatocellular carcinoma (HCC),8 patients with metastase,5 patients with cholangiocarcinoma(CCC),and 9 patients with hemangioma.The shear wave velocity of the tumor and background liver parenchyma were calculated,and results were compared with 30 healthy subjects.Statistical analysis was performed on the shear wave velocity for differentiation of normal liver,background liver parenchyma,and tumors.ResultsHCC and CCC had greater stiffness than metastase (P ＜0.05),there were no statistical differences between HCC and CCC (P = 0.179).Malignant liver tumors had significantly greater stiffness than hemangioma and normal liver (P = 0.000).34.5% (9/26) HCC and 33.3% (4/12) hemangioma appeared softer than the background liver.With a cut-off value of 1.5 m/s for the shear wave velocity,the sensitivity,specificity,positive predictive value and negative predictive value for malignancies were 79.5%,83.3%,94.5% and 52.6%,respectively.ConclusionsARFI elastography quantification is a promising noninvasive technique for assessing solid liver tumors.Use of ARFI elastography quantification may lead to new quantitative tissue characterization parameters for differentiating hemangioma and malignant liver tumors.

Objective To evaluate surgical treatment of lower limb ischemia due to combination of thromboangiitis obliterans (TAO) and arteriosclerosis obliterans (ASO). Methods Clinical data of six patients suffering from lower limb ischemia due to TAO and ASO were retrospectively analyzed. All patients had a history of smoking, and complained of rest pain. Three patients had foot ulcer and one had toe gangrene. Two patients reeeived aortie artery endartereetomy combined with FIFE graft bypass to deep femoral artery and below knee popliteal artery plus saphenous vein graft bypass to tibial artery. One patient received aortic thrombectomy and endarterectomy plus aortofemoral and femoropopliteal PTFE graft bypass. One patient with a history of left graft thrombosis after aorto-bifemoral arterial bypass using PTFE graft before admission underwent left PTFE graft thrombectomy via left groin and left deep femoral artery endarterectomy followed by femoropopliteal arterial bypass. One patient received left common iliac artery endarterectomy plus left profound femoral PTFE graft-left anterior tibial arterial bypass using in situ great saphenous vein graft. One patient received right to left femorofemoral arterial bypass using PTFE graft to left posterior tibial artety bypass using reversed great saphenous vein graft. Results One patient receiving aortoiliac thrombectomy and endarterectomy with aortofemoropopliteotibial arterial bypass suffered from graft thrombosis several hours after operation and emergency thrombectomy with distal posterior tibial arteriovenous fistulization was performed. The rest pain disappeared in 5 patients and partially relieved in one. All the grafts were patent on discharge. The foot ulcer reduced in 3 patients. All the 6 patients were followed up with a mean of 6. 5 months. 3 foot ulcers healed. One patient eventually received below knee amputation due to foot gangrene three months later. Conclusion Although it is difficult to treat combined limb ischemia of TAO and ASO, satisfactory results could be achieved when proper surgical procedure is adopted.

This study aimed to investigate the reconstruction of the thumb and finger extension function in patients with middle and lower trunk root avulsion injuries of the brachial plexus.From April 2010 to January 2015,we enrolled in this study 4 patients diagnosed with middle and lower trunk root avulsion injuries of the brachial plexus via imaging tests,electrophysiological examinations,and clinical confirmation.Muscular branches of the radial nerve,which innervate the supinator in the forearm,were transposed to the posterior interosseous nerve to reconstruct the thumb and finger extension function.Electrophysiological findings and muscle strength of the extensor pollicis longus and extensor digitorum communis,as well as the distance between the thumb tip and index finger tip,were monitored.All patients were followed up for 24 to 30 months,with an average of 27.5 months.Motor unit potentials (MUP) of the extensor digitorum communis appeared at an average of 3.8 months,while MUP of the extensor pollicis longus appeared at an average of 7 months.Compound muscle action potential (CMAP) appeared at an average of 9 months in the extensor digitorum communis,and 12 months in the extensor pollicis longus.Furthermore,the muscle strength of the extensor pollicis longus and extensor digitorum communis both reached grade Ⅲ at 21 months.Lastly,the average distance between the thumb tip and index finger tip was 8.8 cm at 21 months.In conclusion,for patients with middle and lower trunk injuries of the brachial plexus,transposition of the muscular branches of the radial nerve innervating the supinator to the posterior interosseous nerve for the reconstruction of thumb and finger extension function is practicable and feasible.