Abstract:Objective:To establish a human hepatocellular carcinoma cell line derived from portal vein tumor thrombus and study its biological characteristics.Methods: Specimens of liver cancer and its metastasis-portal vein tumor thrombi (PVTT) were freshly resected during operation.The tissues were primary cultured by collagenase Ⅰ digestion method and proliferated.The cell line derived from PVTT grew well.The biological characteristics of the cells were studied by light microscopy,electron microscopy,chromosome analysis,and transplantation experiment.Results: The cell line derived from portal vein tumor thrombus,named CSQT-1,has been cultivated for one year and sub-cultured for more than 100 passages in vitro.The cells either grew as compact colonies (in clusters) or as a monolayered sheet,with a doubling time of 48 h,and exhibited a typical malignant epithelial morphology.The median range of chromosome number was 87-90.Tumor nodes were observed under the skin of nude mice after subcutaneous xenograft transplantation for a month.The established cell line stably expressed CD133.Conclusion: We have successfully established a human hepatocellular carcinoma cell line from PVTT.

Abstract:Objective:To investigate the effect of normoxic acidosis on HIF-1α protein expression and activity in human liver cancer cell line HepG2,so as to assess the role of acidosis in tumor growth.Methods:HepG2 cells were cultured in vitro; the cell growth was assessed by MTT assay after exposure to normoxic acidosis culture media (AP,pH 6.5) for 20 h,and the results were compared with those of the control cells (SD,pH 7.2).Nuclear extractions were performed in the cultured cells,and HIF-1α protein level and HIF-1 DNA binding activities were examined by immunocytochemistry,Western blotting, and HIF-1α Transcription Factor Assay Kit (TransAMTM).The protein level of VEGF (vascular endothelial growth factor) was detected in whole cell extractions by Western blotting assay.Results:MTT assay demonstrated that the cell survival rate was (98.71±1.79)% after treatment with normoxic acidosis culture media (AP,pH 6.5).In comparison with SD group,we noted a slight decrease of cell viability in AP group (P>0.05).Western blotting analysis showed that the levels of HIF-1α and VEGF protein increased by (9.34±1.67) folds and (3.42±0.83) folds,respectively (P<0.01).Moreover,normoxic acidosis greatly enhanced HIF-1 DNA-binding activity compared to SD groups(P<0.01).Conclusion:Normoxic acidosis can evidently increase HIF-1α protein level and DNA-binding activity in HepG2 cells.Furthermore,the protein level of VEGF,an important target gene of HIF-1,is also increased.Therefore it is suggested that,in addition to hypoxia,acidosis may also play an important role in induction of HIF-1 in cancer.

Abstract:Objective:To investigate the correlation of serum adiponectin and hepatic adiponectin receptor(adipoR)expression with pathological changes of the liver in the rats with non-alcoholic fatty liver diseases(NAFLD).Methods: The NAFLD model was induced by an oral administration of high fat diet.The rats were sacrificed at 2,4,8,and 12 weeks.ELISA was used to measure the serum adiponectin and other biochemical parameters.The liver index was also examined.AdipoR mRNA expression in the liver were measured by RT-PCR.Liver slices were observed with Sudan Ⅲ staining,H-E staining and Masson staining for pathological changes.Results: The serum adiponectin in the model group were gradually decreased during the 2nd,4th,8th,and 12th week,and were all significantly lower than those in the control group at corresponding time points(P＜0.01).Serum adiponectin level was found negatively correlated with the liver index(r=-0.383,P=0.015),hepatic inflammation scale(r=-0.475,P=0.002),and hepatic fibrosis scale(r=-0.353,P=0.025).The hepatic adipoR1 mRNA expression in the model group was gradually increased (P＜0.01 from the 4th week) and adipoR2 mRNA expression was gradually decreased compared with the control group (P＜0.01 from the 2nd week). AdipoR2 mRNA expression was negative correlated with liver index(r=-0.431,P=0.006) and hepatic fibrosis scale(r=-0.353,P=0.025).Conclusion: The hepatic adipoR mRNA expression is abnormal in rats with NAFLD; the serum adiponectin level is decreased and negatively correlated with liver inflammation and fibrosis scale,indicating that the decreased serum adiponectin level,pathological hepatic adipoR expression,especially the decreased AdipoR2 expression in the liver may be related to the pathogenesis of NAFLD.

Abstract:Objective:To observe the expression of epidermal growth factor (EGF) and its receptor (EGFR) in the body fluids and renal tissues of rats with autosomal dominant polycystic kidney disease (ADPKD) and to explore their roles in the development of the disease.Methods:EGF levels in the blood and urine of Han:SPRD rats were determined by ELISA. Western blotting,in situ hybridization and immunohistochemistry were used to examine the expression of EGF/EGFR in the kidneys of ADPKD rats. Results:The concentration of EGF in the urine was higher than that in the blood. EGF level in the body fluids of 3 weeks old rats was higher than that of the 3 months old rats. The urine EGF level of cy/+ rats was significantly lower than that in the normal rats (\[263.45±48.53\] pg/ml vs \[321.09±74.57\] pg/ml,P<0.05). The renal distribution pattern of EGF remained unchanged in rats with ADPKD,but the expression level was down-regulated. EGFR in ADPKD rats was ectopically overexpressed on the apical and basolateral membrane of cystic cells. The order of EGFR and phosphorylated EGFR expression was: cy/cy rat> cy/+ rat> +/+rat.Conclusion:The abnormal EGF/EGFR might play a key role in the development and progression of the ADPKD.

Abstract:Objective:To investigate the characteristics of renal papillary calcareous deposits and their role in the formation of calcium oxalate kidney stone,and to explore the formation mechanism of renal calcareous deposits.Methods: A total of 21 patients with calcium oxalate kidney stones were included in the present study.The components of the calculi were detected by Fourier-transform infrared spectroscopy.The calcific plaques were observed and the renal papilla biopsy specimens were obtained during PCNL.The specimens were then subjected to alizarin bordeaux staining and light microscopic examination.The expression of osteopontin,BMP-2, and type Ⅱcollagen in the kidneys was examined by immunohistochemistry.Seven resected renal specimens from patients with non-urolithiasis served as control.Results: Renal papillary calcific plaques were found in all the 21 patients with renal calculi.Local calcareous deposits were found in the renal interstitium around tubular basement membrane and extended toward the mucous membrane in the renal papillas.Microscopically,once the calcareous deposits pierced into the collection system,tiny stones could be seen growing on the deposits.Immunohistochemistristry showed the renal tissues of renal stone patients and normal renal tissues both expressed osteopontin,but not BMP-2 or typeⅡ collagen.Conclusion: Renal papillary calcareous deposits may be one of the initiating nidi for kidney stone formation and they may not be an osteoblastic reaction resembling arteriosteogenesis.

Abstract:Objective:To isolate and screen active marine microorganisms from the East China Sea and to identify the phenotypes of the isolated strains.Methods: Thirty strains of bacteria isolated from the East China Sea were selected randomly,and the active strains were screened out by the anti-Escherichia coli model.The active strains were subjected to physiological and biochemical characteristics,salt-aggregation test,and 16S rDNA sequence analysis.Neighbor-joining trees were constructed by comparing the results of 16S rDNA sequences with sequences described in the BLAST server of the National Center for Biotechnology Information (NCBI); the strains were subsequently identified to genus level.Results: Strains F81612 and F201721 were screened out with the optimum salinities of 10% and 7.5%,respectively.Their morphology and biochemical characteristics were similar to those of Bacillus sp..16S rDNA sequence analysis showed that sequences of F81612 had a higher similarity to those of Bacillus subtilis; F201721 was similar to Bacillus amyloliquefaciens.Conclusion: Two Macrolactin S-producing strains have been screened out by the anti-Escherichia coli model,and they are identified as moderate halophilic Bacillus sp.

Abstract:Objective:To prepare rh-endostatin loaded poly(lactic-co-glycolic acid)(PLGA)microspheres and to evaluate their release behavior in vitro.Methods: Rh-endostatin PLGA microspheres were prepared by W/O/W process.The content and in vitro cumulative release was determined by a HPLC method.Results: The prepared microspheres were well-shaped, with a mean diameter of 122.7 μm.The drug loading and encapsulation efficiency were 1.28% and 38.65%, respectively.The rh-Endo solution (250 μg/ml) showed good stability after placed at 4℃ and 25℃ for 108 h.The cumulative in vitro release was up to 67.37% in 28 days.Conclusion: The rh-Endo can be encapsulated in microspheres to yield sustained release using biodegradable polymers PLGA as the carrier material.

Abstract:Objective:To observe the resection range of dog hemi-vertical larynx by semiconductor laser.Methods: Twelve dogs were randomly divided into three groups (A,B and C,n=4).Group A: the hemi-vertical larynx was cauterized forward from the anterior commissure to thyroid cartilage internal membrane,outwards including the right vocal cords.Group B: the hemi-vertical larynx was cauterized forward from anterior commissure to thyroid cartilage internal membrane and partial cartilage of anterior commissure,outwards including the right ventricular fold,vocal cord,thyroarytenoid muscle and partial lateral cricoarytenoid muscle,downwards including the lower edge of thyroid cartilage,and backwards including the partial arytenoid cartilage.Group C: the hemi-vertical larynx was cauterized forward including the anterior commissure and partial thyroid cartilage of anterior commissure,outwards including the right ventricular fold,vocal cord,thyroarytenoid muscle,lateral cricoarytenoid muscle and paraglottic space to thyroid cartilage,downwards including the partial cricothyroid membrane to the upside of cricoid cartilage,and backwards including total right arytenoid cartilage.The recovery of the laryngeal wounds,hoarseness,and complications were observed and evaluated by using digital camera and electrolaryngoscope immediately,1 week and 4 weeks after operation.The treatment outcome of the resection was evaluated.Results: The laser surgery was completed successfully in all the animals.Laryngoscope showed that the neonatal membrane covered the wound in group A and B one week later,without obvious complications.The neonatal membrane covered part of the wound in group C,accompanied by inflammatory reaction and cough when eating.Four weeks later the new membrane over the wound surface were smooth in all the three groups; adhesion appeared in the anterior commissure and new anterior commissure was formed.A new vocal cord was formed in group A; only a small gap was seen when the glottis was closed.In Group B,the new vocal cord was narrower and thinner; granulation growth was seen in local areas; and the glottis was closed incompletely.In Group C,mucosa was slightly raised over the original location of the vocal cord.Closure of the glottis was poor.Group A had no hoarseness,Group B had moderate,and Group C had severe hoarseness,but without cough when eating.Also in Group C the lamina of thyroid cartilage of the operative side was thinner than that of the contralateral side.Conclusion: When cauterizing dog partial hemi-vertical larynx using semiconductor laser,we can resect all the soft-tissue internal the thyroid cartilage,including the arytenoid and partial thyroid cartilage in the anterior commissure.The repair is satisfactory after operation,without obvious complications.

Abstract:Objective:To study the relationship between the lung compliance and the geographic factor in healthy Chinese adults,so as to provide scientific evidence for establishing the standard value of lung compliance for healthy Chinese adults.Methods and Results: We colleted the lung compliance data from 4 489 (2 726 males,1 763 females)healthy adult Chinese from 46 cities and counties across China.The relationship between lung compliance data and geographic factors (altitude\[X1\],annual sunshine duration\[X2\],annual mean air temperature\[X3\],annual mean relative humidity\[X4\],and annual precipitation \[X5\]) were assessed by correlative analysis and co-linearity diagnostics.Then we analyzed the relationship between the reference value and the geographical factors by using the method Principal Component Analysis.The regression equation was:＝0.079-0.000 001 66X1-0.000 032 3X2-0.002 21X3+0.001 47X4-0.000 017 4X5±0.021.Conclusion: If geographical factors of an area are obtained,the reference value range of healthy adult lung compliance of this area can be calculated by using the regression equation.

Abstract:Objective:To observe the L-Arg(L-arginine) transport,the nitric oxide (NO) production,and NO synthase (NOS) activity in platelets,investigate the significance of the L-Arg-NO system in the pathogenesis of microvascular angina(MVA),and to study the reversing effects of intravenous L-Arg infusion on L-Arg transport.Methods:The 3H-L-Arg transport,NO production,and NOS activity in platelets were examined in 15 patients with MVA and 15 healthy controls.The 15 patients were given intravenous L-Arg infusion (20 g/d) after basic physical examination and were examined again 10 days later.Results:The L-Arg transport in platelets of MVA patients was obviously lower than that in the normal group; the maximum transport velocity (Vmax) decreased by 34.4% compared with the normal group (P<0.01); and the Michaelis constant (Km) increased by 21.4% (P<0.05).The production of NO-2 and the activity of NOS in platelets were decreased by 47.1% (P<0.05) and 25.4% (P<0.05) compared with the normal group,respectively.Intravenous L-Arg infusion reversed the above changes in MVA patients; it increased the Vmax by 11.9% (P<0.01) and decreased Km by 18% (P<0.05); it also increased production of NO-2 by 1.33 folds (P<0.05) and NOS activity by 1.2 folds (P<0.05).Especially,the attack of angina and patient ECG were greatly improved after intravenous L-Arg infusion.Conclusion:L-Arg-NO pathway is impaired in MVA patients,which might be responsible for the endothelium-dependent vascular relaxation in MVA patients.Intravenous L-Arg infusion may benefit the impaired function of L-Arg-NO transport in patients with MVA.

Abstract:Objective:To observe the influence of anisodamine on relaxation function of isolated Wistar and spontaneously hypertensive rat(SHR) aortic rings,and to investigate the underlying mechanism.Methods: The SHR aorta rings were irrigated with anisodamine and the effect of anisodamine on relaxation of isolated aortic rings was observed in normal and spontaneous hypertension rats before and after pretreatment with phenylephrine (PE) or L-nitroarginine methylester(L-NAME).Results: The accumulated concentration of anisodamine showed significantly different relaxing effects on PE-precontracted aortic ring with or without endothelium (P<0.05) in normal rats.Anisodamine showed obvious relaxing effect on PE-precontracted aortic ring with and without endothelium,and the relaxing effect was in a dose-dependent manner,with the maximal relaxation being (78.6±6.9)% and (65.76±11.39)%,respectively (P>0.05).Pretreatment with non-selective NOS inhibitor L-NAME significantly inhibited the relaxing effect of anisodamine on the aortic ring with endothelium (P<0.05).L-NAME partially blocked the relaxing effect of anisodamine on SHR rats,with the maximal relaxation being 61% and 11.9% (P<0.01) before and after blocking,respectively.Conclusion: Anisodamine can enlarge isolated aorta of rats through endothelial and smooth muscle.

Abstract:Objective:To study the differences of activated brain areas during brain error monitoring in schizophrenia patients.Methods: The latencies and amplitudes of error related negativity (ERN) were obtained in 16 patients with first episode schizophrenia and 25 normal controls by high-density event related potential (ERP) technique,and the activated areas were analyzed by low resolution electromagnetic tomography (LORETA).Results: The correct response rate was significantly lower in the schizophrenia participant group than in the control group (P<0.01),and the reaction periods of correct responses and incorrect responses were significantly longer in the schizophrenia group than in the control group (P<0.05,P<0.01).The ERN latencies in the schizophrenic patients were significantly longer on Cz and Pz electrodes compared with those in the control subjects (P<0.05),and ERN amplitudes were significantly smaller than those in the controls (P<0.05).The activation of insula,superior temporal gyrus,middle temporal gyrus,and inferior parietal lobule were obviously lower in the schizophrenic participants than in the controls.Conclusion: Some brain area,such as insular,may contribute to the dysfunction of error monitoring in schizophrenia patients.

Abstract:Objective:To compare the clinical and imaging characteristics between central nervous system vasculitis(CNSV)and mitochondrial encephalomyopathy (ME),so as to analyze the differential diagnosis of the two disorders.Methods: Clinical data on seven patients with CNSV and five with ME were retrospectively analyzed.The clinical manifestations,laboratory parameters,imaging features and histological characteristics were compared to screen for the evidence of their differential diagnosis.Results: The MRI results of both CNSV patients and ME patients (MELAS type) showed a multi-lesion pattern.The symptoms of CNSV patients included headache,limbs weakness,and erythrocyte sedimentation rate (ESR) increase.The symptoms of MELAS patients included epilepsy and increased serum lactic acid.The electroencephalographic manifestations of both diseases were abnormal:CNSV patients mainly had diffused lesions accompanied with limited alterations; ME patients had evidence of epileptic discharge,which was consistent with the clinical symptoms.Conclusion: Clinical manifestations of CNSV and ME patients are more valuable than imaging findings in the diagnosis of the two diseases.CNSV is characterized by vascular disorders and inflammatory reactions; ME is characterized by abnormal energy metabolism and severe damage of gray matter.The final diagnosis should depend on laboratory and histological examinations.

Abstract:Objective:To evaluate the clinical characteristics of Chinese patients with distal hereditary motor neuropathy (dHMN).Methods: The clinical data on patients with dHMN collected from literatures published in China and from the patients treated in our hospital were retrospectively analyzed.Results: The male to female ratio of patients was 1.321. The age of onset was from 13 to 60 years old,with a mean of (38.2±10.4) years old. The incidences of onset from the lower limbs,the upper limbs, and the four limbs were 84.7%,5.6%, and 9.7%,respectively. The incidences of decreased tendon reflexes,lost tendon reflexes in the upper limbs,decreased tendon reflexes and lost tendon reflexes in the upper limbs were 30.4%,65.3%,13.0%, and 87.0%,respectively. The incidences of grade 3-4,grade 2,and grade 0-1 muscle strength of the upper limbs,grade 3-4,grade 2, and grade 0-1 muscle strength of the lower limbs were 55.4%,37.5%,7.1%,50.0%,37.5%, and 12.5%,respectively. No sensory disturbance was found in the patients. Electromyography(EMG)of 26 patients showed neurogenic damages; the motor and sensory nerve conduction velocities were all normal. Eight patients showed neurogenic amyotrophy in muscular biopsy.Conclusion: The onset of dHMN is mainly at middle-age and mainly in the lower limbs. The major manifestations include weakness and atrophy of distal limb muscles,decreased or lost tendon reflexes without sensory loss. Electrophysiology and pathology examinations play an important role in the diagnosis of dHMN.

Abstract:Objective:To evaluate the safety and effectiveness of transcatheter closure of secundum atrial septal defects (ASD) in patients aged over 40 years old.Methods: A total of 115 patients with secundum ASD,aged 40 or over,who were treated by transcatheter closure in our hospital from Oct.1998 to Jul.2005,were enrolled in this study,including 106 with single ASD,eight with multiple ASD and one with complicated patent foramen ovale.All the 115 patients received ASD closure with the AGA Amplatzer device or ASD occluder made in China.The patients received electrocardiogram (ECG),X-ray and transthoracic echocardiography (TEE) before and three days after operation.Follow-up were done 6 months and 3 years after operation.Results: The transcatheter procedure was successfully done in 114 (99.13%) of the 115 patients.The one failed case was transferred to the thoracic surgery department for ASD repair.The complication rate was (3.5%).Classification of heart function (NYHA) improved from 2.11±0.78 to 1.34±0.56 after operation (P<0.001).ECG showed that the right bundle branch block (RBBB) disappeared or improved (complete RBBB to incomplete RBBB).X-ray showed a decrease of cordis/thorax ratio (CTR).TTE displayed that the right venticle (RV) volume was significantly decreased (P<0.001) and ejection fraction was significantly increased (P<0.001).There were three cases of transseptal residual leakage.Conclusion: Transcatheter closure of secundum ASD in patients aged over 40 is safe and effective when indications are strictly followed.

Abstract:Objective:To evaluate the effect and safety of low-dose naloxone combined with sufentanil and ropivacaine for postoperative patient-controlled epidural analgesia (PCEA) in elderly patients undergoing total hip replacement. Methods: A total of 60 elderly patients (ASAⅠ-Ⅱ) undergoing total hip replacement were randomly assigned to two equal groups: Group C was given 0.5 μg/ml sufentanil in 0.15% ropivacaine; Group N was given the same solution with 0.09 μg/(kg·ml) naloxone. The 2 groups were followed up in a double-blinded manner: the visual analogue scale (VAS) for pain score was recorded at 2, 6, 12, and 24 h to evaluate the analgesia effect. Meanwhile, the incidences of postoperative nausea and vomiting (PONV), pruritus, sedation, respiratory depression, and hypotension were also recorded.Results: The VAS pain score in Group N was significantly lower than that in Group C at 6, 12 and 24 h (P<0.01). The incidence of PONV in Group N was significantly lower than that in Group C (P<0.05); the incidences of other opioid-induced side-effects were not significantly different between the 2 groups. During the course of analgesia, the vital signs of 2 groups were stable; no patients had respiratory depression or hypotension.Conclusion: Low-dose epidural naloxone can enhance the analgesic effect of sufentanil. With timely postoperative monitoring, low-dose naloxone combined with sufentanil and ropivacaine is safe and effective for postoperative patient-controlled epidural analgesia in elderly patients undergoing total hip replacement.

Abstract:Objective:To clone human PKGⅠα gene and construct a recombinant adenovirus vector containing wild-type PKGⅠα.Methods: RT-PCR was used to amplify the full-length PKGⅠα gene from human pulmonary arterial smooth muscle.After T/A cloning, PKGⅠα cDNA was cloned into shuttle plasmid pAdTrack-CMV to construct pAdTrack-PKGⅠα.The plasmid was linearized by PmeⅠ and transformed into BJ5183 E.coli, where the plasmid was recombined with pAdEasy-1 by homologous recombination.The recombinants were then transfected into Ad293 cells by Lipofectamine2000 for packaging the adenovirus; the recombinant adenovirus was traced by monitoring GFP expression under fluorescence microscope to determine the titer.Results: PKGⅠα was successfully amplified from human pulmonary arterial smooth muscle by RT-RCR.After 3 cycles of amplification, the titer of adenovirus containing wild-type PKGⅠα reached the indicated level.Conclusion: We have successfully constructed PKGⅠα gene and constructed the PKGⅠα recombinant adenovirus, which provides a foundation for the study of PKGⅠα function and its role in hypoxia pulmonary vessel remodeling.

Abstract:Objective:To investigate the feasibility and outcome of partial hepatectomy and fenestration combined with renal cyst decortication for treatment of autosomal dominant polycystic liver disease (APLD) associated with autosomal dominant polycystic kidney disease (APKD).Methods: From July 2007 to Oct.2007,three patients with APLD associated with APKD were treated by partial hepatectomy and fenestration combined with renal cyst decortication in our hospital.Their preoperative symptoms,operation procedure,operation time,complications and the clinical outcomes were retrospectively analyzed.Results: The 3 patients were successfully treated and discharged.The operation time periods of the 3 patients were 475,402,and 430 minutes.Pleural effusion and ascites occurred in all the 3 patients and disappeared after puncture and drainage.Follow-up was conducted 5,6,and 9 mouths later; the symptoms disappeared in all patients and the renal function became normal.The blood pressure of 2 patients recovered to normal level after operation.Conclusion: Partial hepatectomy and fenestration combined with renal cyst decortication is a safe and acceptable procedure for treatment APLD associated with APKD.The complication rate is comparatively more and the long-term outcomes need to be evaluated further.

Abstract:Glioma is a common tumor of the central nervous system.Tumor cells and tumor microvascular endothelial cells play a very important role in the development and progression of gliomas.Nogo protein is a kind of inhibitory protein in myelin sheaths of the central nervous system and an inhibitor of axonal regeneration in the central nervous system.This paper reviews the relationship between glioma cells and glioma microvascular endothelial cells.

Abstract:Histone deacetylases inhibitors (HDACIs) are a group of compound regulating gene expression at transcriptional level.HDACIs can induce protein hyperacetylation and subsequently lead to chromatin remodeling,cell-cycle arrest,cell differentiation and apoptosis,and transcriptional activation and repression.HDACIs demonstrate promising antitumor activity and are presently tested in clinical studies for solid and hematologic malignancies.Cumulating evidence in animal models of immune disorders also suggests immunosuppressive properties of HDACIs,which provides us with new ideas for treating immune disorders and rejection after transplantation.This paper reviews the recent progress of immunosuppressive function induced by HDACIs from the following aspects:immune diseases,T lymphocytes,inflammatory cytokines and dendritic cells.

Abstract:Psychological stress refers to the process of psychological,physiological function changes induced by perceived threat of the original stress through cognition and evaluation.Psychological stress is almost always accompanied by emotional reactions,so previous studies on influence of psychological stress on the cognitive function were carried out under certain emotional state.Recent studies have shown that stress-induced glucocorticoid increase and catecholamine and glucocorticoid increase caused by stress-induced emotional reactions have different effects on cognitive functions,which is reviewed in this article.

Abstract:Objective:To construct human single-chain variable fragment (ScFv) antibody library,and screen out antibodies against lung adenocarcinoma from the library.Methods: The total RNA was isolated from tumor adjacent lymph nodes of the lung adenocarcinoma patients and was used to amplify VH and VL genes by RT-PCR.VH and VL genes was joined with a DNA linker by SOE-PCR to form the ScFv.The gel purified ScFv gene repertoires were cloned into the phage vector pCANTAB5E to construct the primary phage library.Panning against lung adenocarcinoma cell line A549 was performed for four rounds and the phage library was identified.Results: A recombinant phage antibody library was successfully constructed.The fourth phage harvest yielded 115 times as much as that of the first one.During affinity screening, the antibody was enriched with the increase of panning rounds.Positive reactions to A549 were detected in 7 of 10 randomly selected clones,with a positive rate of 70%.Conclusion: A human phage-display antibody library has been successfully constructed.The selected ScFv fragment can specifically bind to human lung adenocarcinoma cell line A549.

Abstract:

Abstract:目的:观察结直肠三硝基苯磺酸(TNBS)致炎条件下,改变大鼠体内雌激素和孕激素水平对背根神经节(DRG)中P2X3和P2X2 mRNA表达的影响。方法:SD大鼠分为4组:假手术组、去卵巢组、去卵巢加雌激素替代组、去卵巢加雌孕激素替代组,每组手术完成后饲养5周,再行TNBS致炎,5~7 d后用于实验。实时定量RT-PCR法检测各组P2X3 和P2X2 mRNA的表达。结果:去卵巢组大鼠DRG中P2X3 mRNA表达明显低于假手术组,差异有统计学意义(1.08±0.35 vs 1.66±0.66,P<0.05,n=4),雌激素替代组有明显上升(2.06±0.58),明显高于去卵巢组(P<0.01,n=5),而雌孕激素同时替代组明显低于单独添加雌激素组(2.06±0.58 vs 0.87±0.28,P<0.01,n=5)。各组大鼠DRG中P2X2 mRNA表达无统计学差异。结论:结直肠TNBS致炎条件下,雌激素可促进大鼠DRG中P2X3 mRNA表达,但对P2X2 mRNA无明显影响;孕激素的作用可能与雌激素正好相反。炎性条件下,雌孕激素可能通过影响P2X3受体的表达发挥调节炎性痛作用。

Abstract:目的:探讨T细胞共刺激分子CD28在类风湿关节炎(RA)患者外周血和关节液中的表达和意义,同时比较T淋巴细胞亚群CD4、CD8在类风湿关节炎患者外周血和关节液中的表达和意义。方法:选取RA患者45例,平均年龄(47.57±16.43)岁,平均病程(5.84±7.19)年,男性9例,女性36例。取新鲜抗凝外周血细胞(PBMC)应用流式细胞仪进行检测。其中15例有膝关节液者同时抽取关节液单个核细胞(SFMC)行同样检测。与RA年龄和性别匹配的正常体检者54例作为对照。检测CD4、CD8、CD28、CD19(B淋巴细胞)的百分比以及CD4+和CD8+细胞分别表达CD28+的百分比。结果:类风湿关节炎外周血CD4+百分率较正常对照明显升高(P<0.01),但较关节液均明显降低(P<0.05)；外周血CD8+百分率与正常对照无显著差别,而关节液细胞中CD8+百分率较外周血显著升高(P<0.01)。CD28+百分比在关节滑液细胞中较血细胞中有明显增高(P<0.01)。CD4+CD28+/CD4+百分比在类风湿关节炎患者血细胞、关节液细胞与正常对照组均无差别。CD8+CD28+/CD8+百分比在类风湿关节炎关节滑液中较血细胞中有明显增高,二者相差非常显著(P<0.01)。结论:类风湿关节炎患者外周血和关节液中T淋巴细胞亚型和T细胞共刺激分子CD28的表达异常,可能是参与RA发病和病情发展的重要机制之一。

Abstract:目的:评价盐酸二甲双胍缓释片(美哒灵)治疗2型糖尿病的疗效及安全性。方法:2型糖尿病患者120例,随机分为4组：二甲双胍普通片1 500 mg,每次500 mg,每日3次组;二甲双胍缓释片1 500 mg,每日1次组;二甲双胍缓释片1 500 mg,500 mg早1次,1 000 mg晚1次组;二甲双胍缓释片2 000 mg,每日1次组。治疗12周后,观察各组从基线到终点糖化血红蛋白(HbA1c)、空腹血糖(FPG)、总胆固醇(TC)、三酰甘油(TG)的变化以及胃肠道不良反应。结果:113例完成了研究。12周后,所有治疗组的HbA1c均明显下降。二甲双胍缓释片1 500 mg组从基线到终点的平均变化与普通片组之间无明显差异,而2 000 mg组HbA1c下降更为显著(P=0.001)。所有治疗组空腹血糖均明显下降,缓释片组与普通片组的空腹血糖变化接近(P=0.076)。各组总不良反应事件发生率相同,相比普通片组,缓释片组恶心发生率更低(P=0.05)。结论:每日1~2次的二甲双胍缓释片治疗2型糖尿病疗效及安全性与二甲双胍普通片相当,依从性优于二甲双胍普通片。

Abstract:目的:探讨产前超声诊断胎儿小脑延髓池扩张价值及预后。方法:对 2006年6月至2008年1月间来我院进行产前超声诊断检查发现小脑延髓池扩张(>1.0 cm)的胎儿72例进行分类并随访。结果:单纯小脑延髓池增宽60例,小脑延髓池增宽合并其他结构异常6例,Dandy-walker畸形及变异型6例。结论:对于产前超声诊断的小脑延髓池扩张,应准确分类,严密随访,慎重诊断。单纯小脑延髓池扩张并不合并其他系统畸形应定期复查。

Abstract:目的:评价快速上浮脱险后潜艇艇员肾动脉血流动力学的变化。方法:应用多普勒超声对模拟3～30 m快速上浮脱险试验的艇员进行肾动脉血流动力学的监测。结果:3、10、30 m训练深度,双肾动脉阻力指数(RI)、搏动指数(PI)均在正常范围,并随着训练深度的加大变化无统计学意义。结论:浅深度快速上浮脱险对肾动脉血流动力学无显著影响,本次试验的训练方法及保障是安全有效的。

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