A monoclonal antibody (McAb) 3H11 against gastric cancer with high positive rate, high selectivity and high affinity was obtained. 131 I Labeled McAb 3H11 was injected into gastric cancer-bearing nude mice, and the tumor/liver ratio, localization index, ID%/g could reach 8.26?1.26, 6.08 + 1.51, 11.00?2.62, respectively. Moreover, McAb 3H11 conjugated with 131 I could increase the cytotoxicity to tumor xenografts and decrease the side effects. The radioimmunoimage was carried out in 19 cases with gastric cancer, and positive image was obtained in 16 cases (84.2%). The radioimmunoguided surgery of gastric cancer was successfully done by submucous injection of labeled McAb 3H11 through endoscope. The sensitivity, specificity and accuracy for evaluation of tumor infiltrated gastric walls were 94.6%, 96.7% and 95.9%, respectively. The sensitivity, specificity and accuracy for identification of metastatic lymph nodes were 99.2% ,97.7% and 98.8%, respectively.

Objective To evaluate the feasibility of fecal occult blood test(FOBT).combined with colonoseopy,in screening of colorectal cancer.Methods From September 2006 to May 2007.healthv volunteers without any colorectal cancer(CRC)related symptoms in Songjiang District of Shanghai,aging from 40 to 49 or older than 50 years,were recruited.The subjects were firstly screened by FOBT with immunohistochemieal method in two of three consecutive stool samples,and colonoscopy was performed in those with positive FOBT in any of the stool samples.The number.type and distribution of detected polyps were recorded.Results Of 2692 subjects over 50 years old,FOBT was positive in 184(6.8%).in which 1 rectal cancer and 64 polyps,including 34 adenomatous polyps,15 inflammatory polyps and 15 hyperplastic polyps.Of 582 subjects aging from 40 to 49 years,FOBT was positive in 28(4.8%),in which 2 adenocarcinoma and 6 polyps,including 1 adenomatous polyp.Most polyps,including adenomatous polyp and high-risk polyp,were mainly found on the left side of colon.Conclusion CRC screening with FOBT and colonoscopy in asymptomatic average-risk people could detect colon polyps at an early stage.Colonoscopy in patients with positive FOBT would be suitable in large scale CRC screening in communities and rural areas.

BACKGROUND: Establishment of drug (neomycin or hygromycin) resistant feeder layer cells is necessary for screening the target gene positive clone of embryonic stem cell (ESC) transfected in vitro, and the establishment of drug-resistant NIH3T3 cell line is also important for the screening of other target ESC genes.OBJECTIVE: To obtain a feeder layer of positive cell clone of ESC transfected by pTet-on gene by means of the G418-resistant NIH3T3 cell line establishment.DESIGN: Cell culture and DNA examination.SETTING: Faculty of Laboratory Animal, China Medical University.MATERIALS: NIH3T3 cells were contributed by the Cell Biology Staff Room of China Medical University, and pWL/neo plasmid was a gift from Professor Jin Zhuang of Harvard University Medical College. G418,leukemia inhibitory factor (LIF, ESC GRO 106 U/mL) and DMEM were produced by GIBCO BRL Company, while mitomycin-C (MIT-C), DIG marking and kits were the products of Roche Company. Lipoetin was the product of Invitrogen Company, and bought from Shenyang Lianxing Biotechnology Co.,Ltd.METHODS: ①The pWL/neo plasmid containing neo gene was purified and transfected into NIH3T3 cells by lipoetin.②The transfected cells were further cultured in 25 mL medium containing G418 antibiotic of gradient concentration to observe the survival and apoptosis of cells and measure G418 minimal fatal dose (MFD) to NIH3T3 cells.③The transfected cells were subcultured, and the clone of single cell was selected to 24-pore culture board for screening amplification. At the same time, normal NIH3T3 cells were also selected and added with the selective culture medium at the same dose of G418, as screening negative control. ④The cells were planked with lower cell density, and further screened in DMEM medium containing G418 MFD; Meanwhile, normal NIH3T3 cells were taken as controls, and cultured by G418 contain MFD and by DMEM medium without G418 respectively. Light microscope was used to observe G418R NIH3T3 cells. ⑤G418R NIH3T3 cells and MEF were all applied as the feeder layer to culture the ES-D3 cells, which were observed by light microscope. Cell DNA was prepared, and evaluated by PCR and southern blot.MAIN OUTCOME MEASURES: ①G418 MFD to NIH3T3 cells. ②screening result of G418R NIH3T3 cells. ③growth of G418R NIH3T3 cells. ④growth of ES-D3 cells on G418R NIH3T3 cells.RESULTS: ①G418 MFD to NIH3T3 cells was defined as 500 mg/L.②Under the condition of G418 (500 mg/L), G418R NIH3T3 cell clones were successfully selected. ③The G418R NIH3T3 cells had no difference from normal NIH3T3 cells in morphology and propagation rate.④The ESC cultured in feeder layer present a colony growth, smooth limitation and remained an undifferentiation state. The resistant cell genomic DNA of G418R NIH3T3 cells was assayed with specific neo gene primer, and then neo gene DNA fragment was amplified; Southern blot analysis showed that neo gene fragment integrated into G418R NIH3T3 cells.CONCLUSION: The G418R NIH3T3 cells are established successfully,and the ESC cultured in the G418R NIH3T3 cell feeder layer can keep the characteristics of normal ESC.

Objective To construct a 5-lipoxygenase (5-LO) transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different(P<0.05).Conclusion A 5-LO transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.

Objective:To establish a direct dilution-inductively coupled plasma mass spectrometry method for the determination of manganese in urine.Methods:Using 1% nitric acid solution as diluent, the urine dilution factor and internal standard elements were determined by single factor rotation experiment. The linear range, correlation coefficient, precision, accuracy and detection limit of the direct dilution-inductively coupled plasma mass spectrometry for the determination of manganese in urine were evaluated.Results:The linear range of this method was 0.0-20 μg/L, the correlation coefficient was 0.999 9, the detection limit was 0.02 μg/L, the recoveries were 84.65%-103.40%, the relative standard deviations were 0.26%-8.17%.Conclusion:This method has the advantages of simple operation, high sensitivity and low detection limit. It can be used for the determination of urine manganese at the same time with other elements. It is suitable for the determination of urine manganese in workers and ordinary people.

Objective:To establish a direct dilution-inductively coupled plasma mass spectrometry method for the determination of manganese in urine.Methods:Using 1% nitric acid solution as diluent, the urine dilution factor and internal standard elements were determined by single factor rotation experiment. The linear range, correlation coefficient, precision, accuracy and detection limit of the direct dilution-inductively coupled plasma mass spectrometry for the determination of manganese in urine were evaluated.Results:The linear range of this method was 0.0-20 μg/L, the correlation coefficient was 0.999 9, the detection limit was 0.02 μg/L, the recoveries were 84.65%-103.40%, the relative standard deviations were 0.26%-8.17%.Conclusion:This method has the advantages of simple operation, high sensitivity and low detection limit. It can be used for the determination of urine manganese at the same time with other elements. It is suitable for the determination of urine manganese in workers and ordinary people.

Objective:To analyze the endoscopic characteristics of early gastric cancer and precancerous lesions after Helicobacter pylori ( H. pylori) eradication. Methods:From May 2019 to June 2022, at Shanghai Sixth People′s Hospital affiliated to Shanghai Jiaotong University School of Medicine, the medical data of patients diagnosed with differentiated early gastric cancer and precancerous lesions were collected. A total of 93 patients with early gastric cancer and precancerous lesions who had previous history of H. pylori infection and had undergone standardized eradication treatment were selected, and their endoscopic characteristics were retrospectively analyzed. Independent sample t-test, chi-square test, and Fisher′s exact test were used for statistical analysis. Results:Among 93 patients with early gastric cancer and precancerous lesions after H. pylori eradication, there were 56 males and 37 females, with an average age of (66.9±8.2) years old. The time after H. pylori eradication was 3.4 years (range 1.0 to 7.0 years). A total of 109 early gastric cancer and precancerous lesions were found, including 79 patients with single lesion and 14 patients with multiple lesions (30 lesions). There were 60 cases with 73 lesions in the early gastric cancer group and 33 cases with 36 lesions in the precancerous group. Among 93 patients, 89 cases (95.7%) were diagnosed with atrophy level above C-2 according to Kimura-Takemoto classification under endoscopy. The long diameter of 109 lesions was (1.38±0.70) cm and the short diameter was (1.04±0.53) cm. A total of 80 lesions (73.4%) were located in the lower 1/3 part of the stomach, and 53 lesions (48.6%) were located in the lesser curvature. A total of 106 lesions (97.2%) were superficial type (0-Ⅱ) under the endoscopy. The long diameter and short diameter in the early gastric cancer group after H. pylori eradication were both greater than those in the precancerous lesion group ((1.54±0.78) cm vs. (1.06±0.35) cm, (1.16±0.58) cm vs. (0.78±0.33) cm), and the differences were statistically significant ( t=3.53 and 3.73, both P<0.001). There was statistically significant difference in the morphological types between early gastric cancer group after H. pylori eradication and precancerous lesion group ( χ2=11.01, P=0.012). The main morphological type of early gastric cancer after H. pylori eradication was superficial depression type (0-Ⅱc), accounting for 45.2% (33/73), while the precancerous lesions were mainly superficial protruded and flat type, both accounting for 38.9% (14/36). Conclusions:After H. pylori eradication, the endoscopic atrophy range of early gastric cancer and precancerous lesions is mostly above C-2. And the lesions are mostly located in the middle and lower 1/3 part of the stomach, long diameter of lesions <20 mm. The main morphological type is superficial type, especially superficial depression type.

Objective:To evaluate the efficacy and safety of direct antiviral drugs based on sofosbuvir in the treatment of chronic hepatitis C in children.Methods:Data of children diagnosed with chronic hepatitis C from May 2017 to December 2019 and received sofosbuvir (SOF, body weight ≥ 17 kg 200 mg/d, <17 kg 150 mg/d)/ledipasvir (LDV, body mass ≥ 17 kg 45 mg/d, <17 kg 33.75 mg/d) or velpatasvir (VEL, body weight ≥ 17 kg 50 mg/d), were collected. The patients were treated with ribavirin (RBV, 15 mg/kg·d) for 12 weeks and followed up for 12 weeks. The main outcome measures were sustained virological response (SVR) at 12 weeks follow-up after drug discontinuation, and the occurrence of adverse events during the treatment period was observed to evaluate the safety of the drug.Results:HCV RNA turned negative in 10 of the 13 children (76.92%), within 2 weeks after antiviral treatment, in 2 cases (15.38%) HCV RNA turned negative in 2-4 weeks, in 1 case (7.69%) RBV was added when HCV RNA did not turn negative in 8 weeks, but turned negative at 10 weeks. At 12 and 24 weeks after discontinuation of antiviral drugs (SVR12), HCV RNA continued to be negative, and at 24 weeks (SVR24), it was also negative. Adverse reactions: headache occurred in 2 cases (15.38%), fatigue in 3 cases (23.08%), no serious adverse reactions occurred in any of the patients.Conclusions:The efficacy and safety of direct antiviral drugs based on SOF in the treatment of children with chronic hepatitis C were good.

In bone tissue engineering, fabrication of scaffold materials that are biodegradable with regenerative functions is one of the most important research fields. Silk fibroin exhibits many favorable characteristics used as scaffold materials. Among them, hybrid silk fibroin/inorganic composites prepared by biomimetic mineralization have better biocompatibility, biomechanical properties, and biodegradability. At the same time, the hybrid silk fibroin/inorganic materials have much better osteoinduction and conduction properties than silk fibroin. Here, the recent advances in the preparation of silk fibroin/silica hybrid materials by combination or biomimetic silicification are reviewed, and the future research prospects of silicification of silk fibroin are discussed.