Effect of RNAi-mediated IGF1R gene silencing on growth, migration, and invasion of hepatocellular carcinoma cells / 中国病理生理杂志

AIM:To investigate the effect of RNA interference (RNAi)-mediated insulin-like growth factor 1 receptor ( IGF1R) gene silencing on the growth , migration, and invasion of hepatocellular carcinoma cells .METHODS:The most effective siRNA targeting IGF1R gene was designed and screened .After lentiviral expression vector pLVX-shR-NA2-IGF1R carrying the most effective siRNA sequence was constructed , it was transfected into 293T cells and packed into pLVX-shRNA2-IGF1R lentivirus.Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R lentivirus to screen the positive clone Huh7 cells and Hep3B cells with the lentivirus .These Huh7 cells and Hep3B cells were cultured to ana-lyze the mRNA level of IGF1R, cell proliferation, cell cycle, cell apoptosis, cell migration/invasion, and the protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL.RESULTS:The mRNA expression of IGF1R in Huh7 cells and Hep3B cells with pLVX-shRNA2-IGF1R lentivirus was significantly reduced .The proliferation of these cells was remarkably inhibited , and the number in G 1 phase was increased significantly .The percentages of apop-totic cells were increased markedly , and the number of cell migration/invasion was decreased markedly .The protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL were decreased significantly compared with the blank control group and negative control group .CONCLUSION:The RNAi-mediated IGF1R gene silencing sig-nificantly suppresses the growth and the malignant biological characteristics of Huh 7 cells and Hep3B cells, which may be involved in the reduced protein levels of the above genes induced by down -regulation of IGF1R expression.

Inhibitory Effect of siRNA Expression Vector Inhibiting IGF2 Gene on the Proliferation of Hepatoma Cell Line Huh-7 / 中国药房

OBJECTIVE:To investigate the inhibitory effect of siRNA expression vector inhibiting human insulin-like growth factor 2(IGF2)gene on the proliferation of hepatoma cell line Huh-7. METHODS:siRNA expression vector pGL3-hAFP-hTERT-siRNA3(“siRNA3”)which inhibited IGF2 gene by dual promoter regulation of recombinant human alpha-foetoprotein(hAFP)and human telomerase reverse transcriptase(hTERT)was transfected into the Huh-7 cell and normal hepatocyte L-02,and then a nega-tive control group(vector pGL3-hAFP-hTERT)and a blank control group were set up. IGF2 mRNA expression was detected by re-al-time fluorescent quantitative polymerase chain reaction 48 h after transfection into the cells in all groups;the activity of the cells by the microplate reader 0,24,48 and 72 h thereafter;and the cell cycle and apoptosis by the flow cytometer 48 h thereafter,and the changes in the protein levels of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in the cell were detected by Western blot. RESULTS:Compared with the negative control group and blank control group,IGF2 mRNA expression in the Huh-7 cell transfected with siRNA3 was obviously weaker;at 48 and 72 h after transfection,the activity of Huh-7 cell signigicantly reduced, Huh-7 cells at G1 phase obviously increased and those at S phase markedly decreased;the occurrence of early,late and total apopto-sis in Huh-7 cells apparently increased,and the protein expression of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in cells significantly weakened,with statistically significance(P0.05). CONCLUSIONS:siRNA which inhibited IGF2 gene by dual promoter regulation of recombinant hAFP and hTERT can specially inhibit IGF2 gene expression and the prolifer-ation of Huh-7 cells,which may be involved with down-regulated protein expression of cell proliferation-associated gene PCNA, cell cycle control-associated genes Cyclin E2,Cyclin D2 and Cdc2 and apoptosis regulation-associated gene Bcl-2 as a result of down-regulated IGF2 mRNA expression and protein expres-sion.

The practice and effects of pre-job comprehensive training of medical postgraduates / 中华医学教育探索杂志

Objective To improve the teaching methods of graduate students, and provide the theoretical basis for other teaching hospitals to extend the preclinical training mode of clinical medicine professional degree students. Methods 90 clinical medical postgraduate students of Grade 2012 were divided into four groups according to their specialized field and 54 hours of pre-job clinical intensive training were conducted at the same time. The training included four modules lectured by teachers with physician-in-charge above title, such as communication between doctors and patients and medical history collection, physical examination, medical record writing as well as theory of knowl-edge. Before and after the implementation of intensive training, these four skills and knowledge of the students were tested and assessed by professional teachers. Relevant data were paired t test, ANOVA and non-parametric tests. Results The test results of patient-doctor communication and patient history collection, physical examination, complete medical records and theoretical examination results of each group after training were improved to be higher than before (P<0.05). And the effect of patient-doctor communication and patient history collection was the most significant. The doctor-patient communica-tion and history acquisition performance of different groups of graduate students had statistically sig-nificant difference (P=0.001). Conclusion Strengthening the preclinical comprehensive training for medical postgraduate can make the students master the basic clinical skills better and more effectively and the corresponding training methods are worthy of popularization to the other teaching hospitals.

Hepatitis B virus X protein promotes insulin-like growth factor II gene expression by inducing hypomethylation of the P3 promoter in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the involvement of hepatitis B X protein (HBx) in promoter 3 (P3)-driven mRNA overexpression of the insulin-like growth factor II gene (IGF-II) and investigate the underlying epigenetic mechanism.</p><p><b>METHODS</b>Levels of P3 and HBx mRNA and status of P3 methylation were analyzed in human hepatocellular carcinoma (HCC) samples, with and without hepatitis B virus (HBV) infection, using quantitative reverse transcription-PCR and bisulfite sequencing. In addition, the levels of P3 mRNA and P3 methylation were examined in HepG2 cells stably overexpressing HBx (HepG2-HBx). Finally, P3 promoter-luciferase constructs were cotransfected into HepG2 cells along with an HBx-expressing plasmid, and the effects of HBx on transcriptional activity and methylation of P3 were analyzed. Statistical analyses of the data were conducted by chi square test, Fisher's exact test, Student's t-test, Marn-Whitney U test, and Pearson's correlation coefficient test.</p><p><b>RESULTS</b>The HBV-positive HCC specimens had significantly higher levels of P3 mRNA than the HBV-negative HCC specimens (-9.59 ± 3.22 vs. -12.97 ± 3.08 delta CT; P=0.006) but significantly lower levels of P3 methylation (mean values for the 17 CpG sites (36.9% ± 15.5% vs. 52.1% ± 19.1%; P=0.025). The P3 transcript abundance was positively correlated with the level of HBx expression and negatively correlated with the level of P3 methylation. The epigenetic results from experiments with the HepG2-HBx cells were similar. Transfection of HBx significantly decreased P3 methylation level and increased its activity.</p><p><b>CONCLUSION</b>HBx expression may promote IGF-II expression by inducing hypomethylation of its P3 promoter in hepatocellular carcinoma.</p>

Primary abdominal aortoduodenal fistula: a case report / 南方医科大学学报

Abdominal aortoenteric fistula (AAEF) is a rare but life-threatening complication of abdominal aortic aneurysms, and is also an uncommon cause of gastrointestinal hemorrhage. We report a case of primary abdominal aortoduodenal fistula in a 74-year-old male patient, whose main clinical manifestations were abdominal pain, hematochezia, and hematemesis. Colonoscopy, abdominal enhanced computed tomography (CT) and three-dimensional reconstruction angiography confirmed the diagnosis. The patient recovered well after emergent surgical management.

Survey of tele-consultation on Internet for care of myocardial infarction carried out by the chest pain center / 中华急诊医学杂志

Objective To study the efficiency of tele-consultation on Internet with transmitting realtime 12-lead ECG carried out by the Chest Pain Center evaluated by the length of time required for the emergency percutaneous coronary intervention (PCI) in patients with ST segment elevation myocardial infarction (STEMI).Methods A total of 435 STEMI patients treated by emergency PCI were divided into the group A (n =98,admitted in 2010),group B (n =114,admitted in 2011) and group C (n =223,admitted in 2012).Data were collected before (2010) and after establishment of the Chest Pain Center (2011 to 2012) including the length of time elapsed from onset of symptoms to the first medical contact (FMC),the length of time required from FMC to the intra-aortic balloon inflated (FMC-2B) and the length of time required from entering the gate of hospital to the intra-aortic balloon inflated (D-2B).Measure data were described with non-normal median and interquartile intervals.Comparisons were made among groups with rank sum test.Results The median time of D2B of three groups were 107,78 and 59 mins in groups A,B and C,respectively.The differences in D2B among three groups were significant (P =0.000).The time of the D2B was shortened significantly because of the patients transferred to the hospital with a variety of ways (P =0.008).However,the length of D2B time was not significantly changed (P =0.846) when patients came to the hospital all on themselves.The median times from symptom onset to FMC in the group A,group B and group C were 112,62 and 78 mins.and the differences among three groups were not statistically significant (P =0.368).The median times of FMC2B in three groups were 287.0,313.5 and 421.8 mins,respectively,and there were no significant differences (P =0.135).Conclusions The establishment of the Chest Pain Center and Internet of things can effectively shorten the duration of D2B in STEMI patients.However,the reduction of time length from the symptom onset to reperfusion must rely on the coordination between communities and health care system.

A meta-analysis of the effect and safety of angiotensin Ⅱ receptor blockers in treatment of portal hypertension in cirrhotic patients / 中华内科杂志

ObjectiveTo evaluate the efficacy and safety of the angiotensin Ⅱ receptor blockers (ARB) in reducing portal hypertension ( PHT) in patients with cirrhosis.Methods PubMed,EMBASE,Web of Science,The Cochrane Central Register of Controlled Trials,Chinese BiomedicalDatabase,ChineseJournals Full-text Database and WanFang Digital Journal Full-text database were searched.Statistical analysis was performed by meta-analysis using RevMan4.2 software.ResultsAmong 8 randomized controlled trials ( RCT) including 282 patients met the inclusion criteria,4 trials were analyzed to compare the ARB with the placebo or no treatment and the other 4 trials were analyzed to compare the ARB with propranolol.Meta-analysis results were as follows.(1) The ARB resulted in more significant hepatic venous pressure gradient ( HVPG) reduction as compared with the placebo or no treatment [ WMD =1.87 mm Hg (1 mm Hg =0.133 kPa),95％CI ( 0.86-2.87 )mmHg,P =0.00003 ].Andthe ARB were similar to propranolol in reducing HVPG [ WMD =0.92 mm Hg,95％ CI ( - 0.41-2.26)mm Hg,P =0.17 ].(2)The ARB led to more significant reduction in mean arterial pressure than the placebo or no treatment [ WMD =8.89 mm Hg,95％ CI( 7.16-10.62)mm Hg,P ＜ 0.00001 ],but they were similar to propranolol had no significant difference.And the ARB had no significant effect on the heart rate of the patients,which was similar to no treatment group ( P ＞ 0.05 ).Whereas,propranolol could greatly decrease heart rate of the patients ( WMD =- 21.25,95％ CI - 25.83-16.68,P ＜ 0.000 01 ).( 3 ) No significant differences were found in serum bilirubin and creatinine levels between the ARB and the placebo or no treatment groups ( P ＞0.05).The rate of nonspecific adverse events was higher in the ARB groups than in the placebo or no treatment groups ( P =0.03 ),but it showed there was no difference between the ARB and propranolol groups (P =0.72).ConclusionThe ARB is effective in reducing portal hypertension in patients with cirrhosis,which is similar to propranolol.Their effects on mean arterial pressure is similar to propranolol without significant effects on hear rate,liver functionand kidney function,and with less nonspecific adverse events.The ARB could become a new choice for the treatment of portal hypertension.

Effect of HBx on transcriptional activity of IGF-II gene P4 promoter / 中国病理生理杂志

AIM: To construct HBx eukaryotic expression vector pEYFP-C1-X and eukaryotic expression vector pGL3-P4 driven by P4 promoter of human IGF-II gene and to investigate the effect of HBx on the transcription activity of IGF-II gene P4 promoter. METHODS: HBx gene and P4 promoters were cloned into pEYFP-C1 and pGL3-basic vectors respectively by gene recombination techniques to construct recombinant plasmids pEYFP-C1-X and pGL3-P4. HepG2 cells were transfected with pEYFP-C1-X and the resistant cell clones were selected by G418. Then methylated pGL3-P4 was transiently transfected into the above cell clones, and the transcription activity of P4 promoter was determined by dual-luciferase reporter assay system. RESULTS: (1) Aim fragments HBx gene and P4 promoter that were cloned were 465 bp and 1 246 bp, respectively and the DNA sequences were accordant with GenBank data confirmed by restricted enzyme digestion and sequencing. (2) HepG2-EYFP-X cells that expressed HBx protein were obtained. (3) Luciferase activity of methylated P4 promoter in the HepG2-EYFP-X was more than that of control cell HepG2-EYFP (P<0.01). CONCLUSION: HBx may enhance the transcription activity of the P4 promoter.

Hyperproliferation of human colon cancer cells in co-cultures of rat regenerating hepatocytes / 国际外科学杂志

Objective To investigate the stimulated proliferation of colon cancer cells in co-cultures of regenerating hepatocytes. Methods Regenerating hepatocytes(24 hours after partial hepatectomy)were obtained by collagenase perfusion of models of rats undergoing 70% liver resection. To determine whether the ratio of human colon cell line SW480 cells to hepatocytes in co-cultures has influence on their interaction,these cells were cultured in ratios of 1: 101:1, or 10: 1. Proliferation capacity was assessed by the percentage of 3 H-TdR incorporation. Expression of epidermal growth factor receptor(EGFR), insulin-like growth factor1 receptor(IGF-1R)and hepatocyte growth factor receptor(c-met) were analyzed by western blot. Results For co-cultured SW480 and hepatocytes in the ratios of 1: 1 and 1: 10, an increase of disintegrations per minute(dpm) occurred after 72 hours' culture, and lasted at 120 hours' culture(P < 0.05). No difference was found between the group with ratio of 10:1 and control group. Protein levels of EGFR and IGF-1R, but not c-met, were significantly increased between culture of 24 hours and 120 hours; however, no change of these receptors was found in the ratio of 10: 1. Conclusions These results imply that co-culturing human colon cancer cells with regenerating hepatocytes leads to increased expression of EGFR and IGF-1R. We conclude that this effect is probably dependent on paracrine stimulation, by which numerous signals from the hepatocytes contribute to the hyperproliferative state of colon cancer cells via up-regulating the responding receptors.

Clinical study on ultrasound-guided or X-ray-guided endoscopic biliary drainage / 中华消化杂志

Objective To compare the successful ratio, efficacy and complications between ultrasound-guided and X-ray-guided endoscopic biliary drainage (EBD). Methods EBD was performed in 62 patients under ultrasound guidance and 54 patients under X-ray guidance. Serum bilirubin, the bile duct diameter and the changes of clinical symptoms were compared before and after the procedure. Results Tube placement was successfully achieved in 54 of 62 patients under ultrasound guidance and 51 of 54 patients under X-ray guidance. The serum direct bilirubin and the common bile duct diameter in patients with ultrasound guidance before and one week after procedure were (205.41±115.27) μmol/L vs. (106.47±82.16) μmol/L and (12.6±7.1) mm vs. (8.5±3.1) mm, respectively, with significant difference (all P values<0.05). Whereas they were (211.14±106.25) μmol/L vs. (110.89±59.47) μmol/L and (13.1±7.0) mm vs. (8.8± 3.2) mm, respectively, in patients with X-ray guidance (P<0.05). No complications such as abdominal pain, fever and elevated amylase were found in patients with ultrasound guidance, while 3 patients (5.9%) with X-ray guidence had above complications. Conclusions X-ray is a most effective method in guidance of EBD. However, ultrasound guidence, which may avoid unfavorable factors such as X-ray radiation and allergic contrast agent, has some advantages including real-time display, mobile convenience and emergency bedside application. It can instead of X-ray in performance of endoscopic nasobiliary drainage and endoscopic retrograde biliary drainage in patients with bile duct stone and mild narrow ducl caused by tumors.

Analysis of emergency medical rescue system of general hospital during spring festival with snow disaster / 中华急诊医学杂志

Objective To explore the strategy of emergency medical rescue of the massive crowd in general hospital during spring festival with snow disaster. Method The clinical data of 20 966 emergency cases were analyzed retrospectively from 22 Jan,2008 to 6 Feb, 2008 with snow disaster, and concerned about the ratio of different diseases, the character of pre-hospital care and the contrast between emergency medical treatment and routine work. Results The accidence of respiratory disease ( 57.3 % ) was followed by gastrointestinal ( 25.5 % ) and trauma (6.2% )during the emergency medical treatment, and surgical trauma, syncope, coma and convulsion were the most common symptoms, also in some conditions, but empties returning was 30.3% . Similar to the above situation, the extremities (56%)and head injury (24%)were most commonly in the hospital emergency department. The incidence of falling accidents was high( 35.7 % ), and two of them were dead due to trauma on died of being trampled, and on the other was electrothermal burn and falling. Conclusions The general hospital is very important in emergency medical treatment, and it should be ready to tackle the emergency disaster, in order to reduce the loss to minimum.

Construction of the vector for fusion protein gene driven by IGF - Ⅱ P3 promoter and its expression in hepatocellular carcinoma cells / 中国病理生理杂志

AIM: To construct the shuttle plasmid vector for thymidine kinase (tk) and EGFP fusion protein gene driven by IGF - Ⅱ P3 promoter, and investigate the specific killing effect of the HSV - tk/GCV system on hepatocellular carcinoma(HCC) cells in vitro. METHODS: Recombinant shuttle plasmid vector was constructed by techniques of genetic recombination and screening, and identified by restriction digestion and sequencing analysis. Then the recombinant shuttle plasmid was transfected into HepG2 and HeLa cells by techniques of lipofectamine transfection and its expression was detected by fluorescence microscope and RT -PCR. Cell killing after ganciclovir(GCV) application was determined by MTT. RESULTS: Identification of pDC316 -tkEGFP- P3 by enzyme digestion and sequencing analysis showed that the length, inserted location and direction of the target genes which were inserted into the recombinant were correct. It was found that enhanced green fluorescence protein could only be seen in HepG2 cells, but not in HeLa cells. The results of RT -PCR showed that only two bands could be seen in the samples of pDC316 -tkEGFP- P3 transfected HepG2 cells. The MTT test showed the selective cytotoxicity of GCV to the transfected HepG2 cells. CONCLUSION: The shuttle plasmid vector carrying the tkEGFP fusion protein gene driven by IGF - Ⅱ P3 promoter has been constructed successfully and its specific expression in HepG2 cells provided a sound basis for targeted gene therapy for HCC.

The Renaturation and Activity Study of LexA From Pseudomonas aeruginosa / 中国生物工程杂志

Objective:To optimize the renaturation procedure of denatured LexA,prepare the repressor LexA from Pseudomonas aeruginosa(PA),which have the satisfactory biologic activity.Methods:The LexA was renatured by the GSH/GSSG dilution method,and the renatured protein were purified by Ni2+ chelate affinity chromatography and gel filtration chromatography,following desalination by Sephadex G25 gel column.The renaturation result were detected by the native polyacrylamide gel electrophoresis and RPHPLC.The immunological activity of all LexA proteins,including the denatured,renatured protein and the renatured protein that was treated with the DTT,were determined by Western blot.Results:The renatured LexA appears both monomer and multimer,which is confirmed by the native polyacrylamide gel electrophoresis analysis and RPHPLC.Gel retardation experiments shows that the renatured LexA have satisfactory biologic activity.

Relationship between alterations of p16(INK4a) and p14(ARF) genes of CDKN2A locus and gastric carcinogenesis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the relationship between alterations of p16(INK4a) and p14(ARF) genes and gastric carcinogenesis.</p><p><b>METHODS</b>The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied. The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16(INK4a) and p14(ARF) genes were assessed by PCR, PCR-SSCP, PCR based methylation assay, and RT-PCR.</p><p><b>RESULTS</b>(1) The homozygous deletion rate of p16(INK4a) and p14(ARF) was 35.4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor. (2) There was no point mutation of p16(INK4a) and p14(ARF) in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor. (3) Methylation of the CpG islands of p16(INK4a) and p14(ARF) was detected in 47.9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference (P < 0.01). (4) The loss rate of p16(INK4a) mRNA was 47.9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1alpha and 2 had a higher loss rate (100%, 6/6) of p16(INK4a) mRNA than those of the methylation of the other exons (11.8%, 2/17, P < 0.01); the loss rate of p14(ARF) mRNA was 45.8% (22/48) in gastric cancer, and patients with the combined methylation of exons 1beta and 2 had a higher loss rate (100%, 3/3) of p14(ARF) mRNA than those of the methylation of the other exons (15%, 3/20, P < 0.05). (5) The combined loss of p16(INK4a) and p14(ARF) mRNAs was examined in 1 (5.6%) of 18 patients of well and moderately-differentiated carcinomas, and 11 (36.7%) of 30 patients of poorly and not-differentiated carcinomas with a significant difference (P < 0.05).</p><p><b>CONCLUSION</b>p16(INK4a) and p14(ARF) genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer.</p>

Cloning of P1 and P3 promoters of human insulin-like growth factorⅡ gene / 中国病理生理杂志

AIM: To clone P1 and P3 promoters of the human insulin-like growth factor Ⅱ(IGF-Ⅱ) gene. METHODS: According to the complete DNA sequence of IGF-Ⅱ gene, the nested primer PCR was performed for amplifying P1 and P3 promoter fragments of the gene from human L-02 cell line.These PCR products were analyzed by agarose gel electrophoresis, and cloned by using TOPO TA Cloning kit. The positive clones containing P1 and P3 fragments were selected and confirmed by sequencing.RESULTS: The DNA sequences of P1 and P3 promoters cloned were accordant with GenBank data. CONCLUSION: In this study P1 and P3 promoters of the IGF-II gene were cloned successfully.

Intraperitoneal chemotherapy for malignant ascites of gastric cancer based on the results of in vitro drug sensitivity test of anticancer agents / 中华消化杂志

Objective To estimate the sensitivity of malignant cells from ascites of gastric cancer to anticancer agents by in vitro sensitivity test, and to study its value in individualized intraperitoneal chemotherapy. Methods Forty-seven cases of gastric cancers with malignant ascites were selected. The gastric cancer cells were isolated from the ascites of 19 patients with gastric cancers, and in vitro sensitivity tests to carboplatin, taxol, fluorouracil, cisplatin, adriamycin, hydroxycamptothecine, methotrexate, mitomycin and dacarbazine were performed by ATP bioluminescence assay. One of the most sensitive drugs was applied to intraperitoneal chemotherapy. The rates of the patients with complete remission of ascites and with disappearance of intraperitoneal cancer cells in the sensitivity test group were compared to those of 28 patients in control group who all were treated with intraperitoneal cisplatin. Results In sensitivity test group, the sensitive cases to carboplatin, taxol, fluorouracil, cisplatin, adriamycin, hydroxycamptothecine, and mitomycin were 7,6,6,6,5,5 and 5 respectively. No case was sensitive to methotrexate or dacarbazine. The rates of patients with complete remmission of ascites and the rates of patients with disappearance of intraperitoneal cancer cells in the sensitivity test group were significantly higher than those in cisplatin control group(68.4% vs. 32.1%, P

Alterations of p14~(ARF) and p53 genes in human primary colorectal carcinomas / 中国病理生理杂志

AIM: To investigate the genetic and epigenetic alterations of p14~ ARF gene and mutation status of p53 gene in human primary colorectal carcinomas and to analyze the relationship between the two gene changes and the role of abrogation of the p14~ ARF -p53 pathway in colorectal carcinogenesis. METHODS: The homozygous deletions, mutations, methylation of 5′ CpG islands, mRNA expression of p14~ ARF gene and mutations of p53 gene were assessed by PCR, direct sequencing, methylation-specific PCR, and RT-PCR in the tumorous and matched adjacent normal colorectal tissues from 56 patients with colorectal carcinoma. RESULTS: ① p14~ ARF alterations were detected in 27% (15/56) of colorectal carcinoma tissues studied, of which 1 case showed homozygous deletion, 14 cases showed 5′ CpG island methylation, and no mutation was found in any tumor. ②15 colorectal carcinomas with p14~ ARF alterations indicated lack of (13 cases) or at low level of expression (2 cases) of p14~ ARF mRNA, while expression of the p14~ ARF transcript was detected in the remaining 41 colorectal carcinomas and any matched adjacent normal colorectal tissues. ③ The mutations of p53 gene were detected in 48% (27/56) of colorectal carcinomas investigated. ④ Of these 56 cases, 12 had p14~ ARF alterations alone, 24 had p53 mutations alone, 3 had both p53 mutations and p14~ ARF methylation, and 17 had neither. 70% (39/56) of the samples had either or both abnormalities of the two genes, and p14~ ARF hypermethylation was related to wildtype p53 (P

Affinity maturation of a single-chain antibody for hepatocellular carcinoma by error-prone PCR and DNA shuffling / 中国病理生理杂志

AIM: To obtain a single-chain antibody with high affinity for hepatocellular carcinoma (HCC). METHODS: A second single-chain antibody mutant library was established by using error-prone PCR and DNA shuffling. Single-chain antibodies with high affinity for hepatocellular carcinoma were selected from phage antibody library by using ELISA. RESULTS: The content of the second single-chain antibody mutant library was about 4.5?10~7. Two selected mutants M25 and M36 were obtained after 3 rounds of panning and ELISA. Immunoassay showed that M25 and M36 bound to human HCC cells specifically. The relative affinity of M25 was 2.0 folds higher than that of the original antibody, and M36 was 2.4 folds higher than the original antibody. CONCLUSION: Error-prone PCR combined with DNA shuffling is an effective method to improve affinity of antibodies isolated from phage antibody library.

Significance and alterations of p14~(ARF) gene of CDKN2A locus in gastric cancer / 中国病理生理杂志

AIM: To investigate the significance and changes of p14 ARF gene in gastric cancer.METHODS: The tumors and gastric tissues neighboring carcinoma from 48 patients with gastric cancer were studied. The homozygous deletions, mutations, methylation of the CpG islands, and mRNA expression of p14 ARF gene were assessed by PCR, PCR-SSCP, PCR based methylation assay, and RT-PCR.RESULTS: ①The homozygous deletion rate of p14 ARF was 31 3% (15/48), and no homozygous deletions were examined in all the gastric tissues neighboring tumor. ②There were no point mutations of p14 ARF in 33 gastric cancers without homozygous deletion and in the matched gastric tissues adjacent to tumor. ③Methylation rate of the CpG islands of p14 ARF was significantly higher in gastric cancers(47.9%, 23/48) than that in gastric tissues neighboring cancer (4.2%, 2/48)( P

Effect of probiotic preparations on inducing and mainfaining remission of ulcerative colitis:a meta-analysis / 解放军医学杂志

Objective To evaluate the effect of probiotics on inducing and maintaining remission of ulcerative colitis (UC). Methods PubMed, EMBase, Web of Science and The Cochrane Central Register of Controlled Trials from 1966 to 2009 (up to August), and China Journal Full-text Database, Chinese Technologic Journal Database (Weipu), Wan Fang Digital Journal Full-text Database from 1978 to 2009 (up to August) were retrieved in order to collect clinical randomized controlled trials regarding the effect of probiotics in remission induction and maintenance in UC patients. Statistical analysis was performed by meta-analysis using Review Manager 4.2.10. Results Eleven randomized controlled trials met the inclusion criteria, of which five studies were included in the control trial of probiotics and placebo to evaluate the clinical remission rate, seven studies were included to compare probiotics with placebo (3 trials) or with mesalazine (4 trials) to evaluate the clinical relapse rate (one of the trials was included in the induction treatment group and the maintenance treatment group). Meta-analysis showed that: a) On the basis of combining with conventional therapy, probiotics were superior to placebo in clinical remission rate (OR=0.28, 95%CI: 0.16-0.49, P