OBJECTIVE: To evaluate the histocompatibility of poly (lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide (PLGNRIP) sustained release microspheres.METHODS: The crude peptide comprising N to C-terminals was synthesized using Fmoc method. The crude synthetic RNAⅢ peptide was purified by reverse phase high performance liquid chromatography, followed by component harvesting according to ultraviolet absorption peak, and freeze-drying. PLGNRIP sustained release microspheres with a diameter of 50-70 pm were prepared using liquid-phase multiple emulsion method. The histocompatibility of PLGNRIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test, MTT cytotoxicity test, intramuscular implantation test, sensitivity test, and pyrogen test.RESULTS: Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass. MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%, with cytotoxicity grade 1, which indicated no cytotoxicity. Intramuscular implantation tests showed that at 4 weeks after implantation of RiP powder or PLGNRIP microscopes, no obviously congested, degenerated, or necrotic tissue was observed. All RIP powder and a part PLGNRIP microscopes were degraded. Fibroblasts accounted for a large proportion in all cells. NO inflammatory cell infiltration, involving neutrophits and multinucleated giant celts, was observed. Sensitivity test rasults displayed that the average primary irritation index was 0.38, 0.33, arid 0.31 in the eluent stock solution, 2% dinitoflruorobenzene, and physiological saline-administerd groups, respectively. Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5 ℃ and the sum of fervescence was under 1.3 ℃ .This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION: PLGNRIP sustained release microspheras exhibit good histocompatibility.

OBJECTIVE: To evaluate blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide (PLGA/RIP) delayed release microspheres.METHODS: ① Preparation of PLGA/RIP microspheres: The solid-phase synthesis (Fmoc) method was used to synthesize RIP crude sample from C end to N end; the synthesized crude peptide was purified by the reverse phase high performance liquid chromatography. According to UV absorption peak, the components were collected and freeze-dried, to obtain RIP purifications. Then liquid-phase multiple emulsion method was used to prepare PLGA/RIP microspheres at the diameter of 50-70 μm. ② Preparation of eluent: The PLGA/RIP microsphere powders were eluted with sterile physiological saline at 37 ℃, to prepare 1 g/L eluent; then 0.5 g/L eluent was obtained adding equal volume of sterile physiological saline. The hemolysis test, blood clotting test, and platelet aggregation test were conducted to measure prothrombin time and activated partial thromboplastin time, to observe the influence on rabbit leucocytes, erythrocytes and thrombocytes, and to preliminarily evaluate the blood compatibility of PLGA/RIP microspheres. RESULTS: ①The haemolysis rates of eluent stock solution and 0.5 g/L eluents were 3.24% and 2.67% respectively, which were in coincidence with the criteria of medical biomaterials, less than 5%. ② The eluent stock solution and 0.5 g/L eluents of PLGA/RIP microspheres had no significant effect on rabbit clotting time, prothrombin time and activated partial thromboplastin time, the number of rabbit leucocytes, erythrocytes and thrombocytes, as well as platelet aggregation.CONCLUSION: PLGA/RIP delayed release microspheres have a good blood compatibility.

BACKGROUND: Staphylococci inhibitor, RNAⅢ inhibiting peptide, has been firstly synthesized in China, while RNAIII inhibiting peptide/bioinspired phosphorylcholine-based cytomembrane coating (RIP/PC) has been also prepared.OBJECTIVE: To evaluate the blood compatibility of RIP/PC.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at Institute of Clinical Pharmacologic Research and Medical Animal Experimental Center of General Hospital of Chinese PLA from October 2005 to October 2007.MATERIALS: A total of 30 healthy adult New Zealand rabbits were provided by Animal Experimental Center of General Hospital of Chinese PLA.METHODS: The 316L stainless steel was dip into a mixture of 5 g/L quaternionic copolymer tetrahydrpfuran and 10% RNAⅢ inhibiting peptide to obtain stable polymer coating, which was extracted at density of 1 cm~2/mL in saline at 37 ℃ for 72 hours to get 100% eluent. The same volume of saline was added to make 50% eluent. The effects of RIP/PC on leukocytes, eryfhrocytes,and blood platelet of rabbits were detected via measuring prothrombin time and activated thrombin time during hemolysis test,hemagglutinatin test, and blood platelet aggregation test.MAIN OUTCOME MEASURES: Hemolytic ratio of erythrocytes, clotting time, prothrombin time, activated thrombin time, amounts of leukocytes, erythrocytes, and blood platelet, as well as blood platelet aggregation.RESULTS: Hemolysis test showed that the hemolytic ratio was 3.08% and 1.88% of 100% and 50% eluent, respectively; both the hemolytic ratios were < 5%, suggesting being coincidence with hemolytic requests of biological materials. The hemagglutinatin test showed that both 100% and 50% eluent did not have any effect on clotting time, prothrombin time, activated thrombin time,amounts of leukocytes, erythrocytes, and blood platelet, as well as blood platelet aggregation.CONCLUSION: The firstly synthesized RIP/PC has a good biocompatibility.

OBJECTIVE:To evaluate the histocompatibility of poly(lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide(PLGA/RIP)sustained release microspheres.METHODS:The crude peptide comprising N to C-terminals was synthesized using Fmoc method.The crude synthetic RNAIII peptide was purified by reverse phase high performance liquid chromatography,followed by component harvesting according to ultraviolet absorption peak,and freeze-drying.PLGA/RIP sustained release microspheres with a diameter of 50-70?m were prepared using liquid-phase multiple emulsion method.The histocompatibility of PLGA/RIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test,MTT cytotoxicity test,intramuscular implantation test,sensitivity test,and pyrogen test.RESULTS:Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass.MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%,with cytotoxicity grade 1,which indicates no cytotoxicity.Intramuscular implantation tests showed that at 4 weeks after implantation of RIP powder or PLGA/RIP microscopes,no obviously congested,degenerated,or necrotic tissue was observed.All RIP powder and a part PLGA/RIP microscopes were degraded.Fibroblasts accounted for a large proportion in all cells.No inflammatory cell infiltration,involving neutrophils and multinucleated giant cells,was observed.Sensitivity test results displayed that the average primary irritation index was 0.38,0.33,and 0.31 in the eluent stock solution,2% dinitoflruorobenzene,and physiological saline-administerd groups,respectively.Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5℃ and the sum of fervescence was under 1.3℃.This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION:PLGA/RIP sustained release microspheres exhibit good histocompatibility.

BACKGROUND: Carbonated hydroxyapatite cement is a new type material for skeletal repair and hydroxyapatites have been applied in the clinical treatment of skeletal defect.OBJECTIVE: To observe the effective characteristics of carbonated hydroxyapatite cement on repair of skeletal defect by animal experiment.DESIGN: Paired design, self-controlled and verified experiment was applied in the research.SETTING: Orthopedic Institute and Animal Experimental Center of Chinese PLA.MATERIALS: The experiment was performed in Orthopedic Institute and Animal Experimental Center of Chinese PLA from May 2002 to January 2003, in which, 10 healthy adult male mongrel dogs were applied, body mass weighted varied from 20 to 22 kg.METHODS: Animal model of skeletal defect was prepared on proximal ends of humeri of 10 mongrel dogs thydroxyapatitet were randomized into experimental side and control side. Ceramics repair of skeletal defect was done by carbonated hydroxyapatite cement and high-temperature sintered hydroxyapatite respectively. The animals were sacrificed on the 5th day, 4th, 8th, 12th and 16th weeks successively after operation. The repair effects were performed with X-ray and histological observation.staining.Results of stereomicroscopic and X-ray observations on bilateral skeletal defect: Osseointegration with carbonated hydroxyapatite cement was tight on the experimental side and the interface became unclear gradually with time lasting. The interface between hydroxyapatite and bone was still clear on the and eosin staining and thydroxyapatitet of ground bone with Gimsa staining:On the 8th week on the experimental side, the new bone grew into carbonated hydroxyapatite cement, on the 16th week, the two parts were intermixed and integrated and the bone island was formed around newly generated vessels in carbonated hydroxyapatite cement. On the control side, hydroxyapatite still maintained integrated and the bone interface was clear between hydroxyapatite and bone. On the 16th week, the aggradation of newly generated bone presented on hydroxyapatite surface.CONCLUSION: Carbonated hydroxyapatite cement possesses solidification property in situ, biocompatibility and osseous conductive activity. It is the satisfactory new type material for repair of skeletal defect.

Objective To investigate and analyze risk factors of re-fracture after operation of osteoporotic hip fracture.Methods Two hundred forty-seven patients receiving operation of osteoporotic hip fracture were retrospectively studied and followed up,and all patients were divided into re-fracture group (54 patients) and no-re-fracture group (193 patients).The related factors such as sex,age,body mass index (BMI),affected side,initial fracture site,operation type,perioperative blood loss,postoperative delirium,postoperative bedridden time,medical complications,Charlson comorbidity index,antiostoporosis therapy,hip function scores with Harris and functional independence measurement (FIM) scores were compared by single factor analysis and multivariate Logistic regression analysis.Results Single factor analysis and multivariate Logistic regression analysis both showed that the risk factors of re-fracture after operation of osteoporotic hip fracture included age,postoperative delirium,hypertension,diabetes mellitus,cerebrovascular disease,antiostoporosis therapy,hip function scores with Harris and FIM scores (P ＜ 0.05 or ＜ 0.01).Conclusions Risk factors of re-fracture after operation of osteoporotic hip fracture include passive factors of age,postoperative delirium and medical complications,and subjective factors of antiostoporosis therapy,hip function scores with Harris and FIM scores.Patients should receive medical treatment positively,enhance antiostoporosis therapy and rehabilitation training of hip function to prevent re-fracture.

Objective:To clarify the clinicopathological, especially molecular, features of early-onset gastric cancer with the aim of informing analysis of treatment strategies.Methods:In this retrospective case-control study, we examined data from a dedicated gastric cancer database in Zhongshan Hospital affiliated to Fudan University. The original cohort comprised 2506 patients with gastric cancer who had undergone gastrectomy in Zhongshan Hospital Fudan University from July 2020 to October 2021, including 198 with early-onset gastric cancer (aged ≤45 years) and 2,308 with non-early gastric cancer. We used a simple random sampling method to select 396 of the 2,308 patients aged >45 years (ratio of 1:2) as the control group and then compared molecular diagnostic data and clinicopathological features of the two groups.Results:The median age was 39 years in the early-onset gastric cancer group, while 66 years in the control group. The clinicopathological features of early-onset gastric cancer included female predominance (59.1% [117/198] vs. 27.8% [110/396], χ 2=54.816, P<0.001), less comorbidity (32.3% [64/198] vs. 57.1% [226/396], χ 2=32.355, P<0.001), poorer differentiation (93.9% [186/198] vs. 74.5% [295/396], χ 2=30.777, P<0.001) and higher proportion of diffuse type (40.4% [80/198] vs. 15.9% [63/396], χ 2=69.639, P<0.001), distant metastasis (7.1% [14/198] vs. 2.8% [11/396], χ 2=6.034, P=0.014). Regarding treatment, distal gastrectomy was more commonly performed than proximal gastrectomy (55.1% [109/198] vs. 47.0% [186/396], 1.5% [3/198] vs. 8.3% [33/396], χ 2=11.644, P=0.003). Family history of gastric cancer, TNM stage, tumor size, lymph node dissection, nerve invasion, nodes harboring metastases, range of lymph node dissection, digestive tract reconstruction procedure, implementation of laparoscopic surgery, combined resection, and preoperative treatment did not differ significantly between the two groups (all P>0.05). Molecular diagnosis showed there was a smaller percentage of mismatch repair deficiency in the early-onset gastric cancer than in the control group (1.0% [2/198] vs. 10.1% [40/396], χ 2=16.301, P<0.001), and a higher rate of positivity for Claudin 18.2 (77.8% [154/198] vs. 53.0% [210/396], χ 2=5.442, P<0.001). HER-2 and Epstein–Barr virus positivity rates did not differ significantly between the two groups. Conclusion:Early-onset gastric cancer is a distinct type of gastric cancer with a high degree of malignancy, and treatment targeting Claudin 18.2 may be effective.

Objective:To clarify the clinicopathological, especially molecular, features of early-onset gastric cancer with the aim of informing analysis of treatment strategies.Methods:In this retrospective case-control study, we examined data from a dedicated gastric cancer database in Zhongshan Hospital affiliated to Fudan University. The original cohort comprised 2506 patients with gastric cancer who had undergone gastrectomy in Zhongshan Hospital Fudan University from July 2020 to October 2021, including 198 with early-onset gastric cancer (aged ≤45 years) and 2,308 with non-early gastric cancer. We used a simple random sampling method to select 396 of the 2,308 patients aged >45 years (ratio of 1:2) as the control group and then compared molecular diagnostic data and clinicopathological features of the two groups.Results:The median age was 39 years in the early-onset gastric cancer group, while 66 years in the control group. The clinicopathological features of early-onset gastric cancer included female predominance (59.1% [117/198] vs. 27.8% [110/396], χ 2=54.816, P<0.001), less comorbidity (32.3% [64/198] vs. 57.1% [226/396], χ 2=32.355, P<0.001), poorer differentiation (93.9% [186/198] vs. 74.5% [295/396], χ 2=30.777, P<0.001) and higher proportion of diffuse type (40.4% [80/198] vs. 15.9% [63/396], χ 2=69.639, P<0.001), distant metastasis (7.1% [14/198] vs. 2.8% [11/396], χ 2=6.034, P=0.014). Regarding treatment, distal gastrectomy was more commonly performed than proximal gastrectomy (55.1% [109/198] vs. 47.0% [186/396], 1.5% [3/198] vs. 8.3% [33/396], χ 2=11.644, P=0.003). Family history of gastric cancer, TNM stage, tumor size, lymph node dissection, nerve invasion, nodes harboring metastases, range of lymph node dissection, digestive tract reconstruction procedure, implementation of laparoscopic surgery, combined resection, and preoperative treatment did not differ significantly between the two groups (all P>0.05). Molecular diagnosis showed there was a smaller percentage of mismatch repair deficiency in the early-onset gastric cancer than in the control group (1.0% [2/198] vs. 10.1% [40/396], χ 2=16.301, P<0.001), and a higher rate of positivity for Claudin 18.2 (77.8% [154/198] vs. 53.0% [210/396], χ 2=5.442, P<0.001). HER-2 and Epstein–Barr virus positivity rates did not differ significantly between the two groups. Conclusion:Early-onset gastric cancer is a distinct type of gastric cancer with a high degree of malignancy, and treatment targeting Claudin 18.2 may be effective.

Objective:To optimize key parameters based on microwave assisted rapid staining technique for ultrathin section and establish a rapid ultrathin section preparing method .Methods:Ultrathin sections were stained respectively with 1% uranium acetate (UA) and lead citrate (LC) for different duration at various microwave power using microwave tissue processor. Then transmission electron microscope (TEM) photographs of cellular ultrastructure were taken and analyzed. The optimized single staining parameters were decided and combined to investigate the optimal microwave assisted UA and LC double staining conditions. The rapid staining effects of ultrathin sections were verified in different viruses including human adenovirus 5 (HAd5), herpes simplex virus (HSV), H1N1 influenza virus, enterovirus 71(EV-A71)human infected cells samples.Results:The optimized microwave assisted rapid ultrathin section staining parameters are: UA for 30 s and LC for 20 s at power 200 W or UA for 30 s and LC for 30 s at 300 W using microwave tissue processor.1∶6 dilution of original LC concentration could still work well through microwave assistance. The parameters can be extended and applied to domestic microwave ovens, and the optimized staining parameters are UA for 30 s and LC for 30 s at 320 W.Conclusions:The optimized parameters of microwave assisted rapid ultrathin section staining were obtained and can be applied in not only cell samples but also different virus ultrathin sections.