OBJECTIVE: To prepare chitosan microspheres encapsulated transforming growth factor β1 (TGFβ1), and to analyze its property. METHODS: The chitosan was dissolved in 2% acetic acid to prepare chitosan microspheres encapsulated TGFβ1 with emulsification cross-linking method, Tween 80 and sodium polyphosphate were served as emulsifying agent and cross-linking agent, respectively. Meanwhile, chitosan microspheres and containing bovine serum albumin chitosan microspheres were prepared as blank control and experimental control groups. The morphology and diameter of 3 kinds of microspheres were observed, and the dispersion and in vitro release of chitosan microspheres encapsulated TGFβ1 were detected, furthermore, the water absorption expansion rate of blank control and experimental control groups were measured. RESULTS: Scanning electron microscopy showed that the microspheres diameter in the blank group was approach 15 μm, with smooth surface and plenty of tiny pores. However, the microspheres in the other 2 groups were distributed uniformly with approximately 1 μm in diameter, the surface was smooth. The chitosan microspheres encapsulated TGFβ1 released fast at begin 12 hours, and then gentled gradually, with 53.5% release ratio within 6 days. The increased mass of microspheres in the blank control and experimental control groups reached a balance after 1 hour, both of which were over 700%,in particular larger in the acid environment. CONCLUSION: Chitosan microspheres encapsulated TGFβ1 prepared by emulsification cross-linking method exhibit high yield and good drug release. The strong water absorption expansion rate of chitosan microspheres requires aperture size, as well as intensity of bone tissue engineered scaffold.

BACKGROUND: Simply used natural materials-prepared scaffolds such as collagen, gelatin and fibrin solve problems of biocompatibility, but its degradation is rapid, and cannot induce new tissues, but collapse is found as cell scaffolds.OBJECTIVE: To explore and determine the property of biological degradable three-dimensional porous scaffolds using silk fibroin-chitosan composite.DESIGN, TIME AND SETTING: The material observational study was performed at the Institute of Bioengineering, Zhejiang Academy of Medical Science from June 2008 to June 2009.MATERIALS: Spring silk cocoon was presented by a silkworm farmer from Huangdunmiao village, Maqiao town, Haining City,Zhejiang Province, China. Chitosan was produced by Shanghai Bo'ao Biological Technology.METHODS: 15 g/L silk fibroin solution was made by degumming, salvation and dialysis. Chitosan was dissolved in 2% acetic acid solution to prepare 25 g/L chitosan-acetic acid solution. Two solutions were mixed to prepare six silk fibroin/chitosan solutions, and mass ratio was 10: 0, 5: 5, 4: 6, 3: 7, 2: 8, 0:10. These solutions were separately sucked in a 24-well plate. Following exhausting gas vacuole at 4 ℃, precooling was performed at -20 ℃ for 12 hours, followed by cryodesiccate for 30 hours. Samples were then hydrated in ethanol, neutralized in NaOH-alcohol for 1 hour, washed and then frozen to dry.MAIN OUTCOME MEASURES: Optical microscope and scanning electron microscope were used to observe pore size and structure of various mass ratio-prepared scaffold. Modified liquid substitution method was utilized to measure porosity of various scaffolds. The degradable rate of various scaffolds was determined at 4 weeks in vitro.RESULTS: Silk fibroin/chitosan of 10: 0 mass ratio-prepared scaffold had rough fluffy pore, was brittle, with high dissolve-loss rates. On the contrary, chitosan-prepared scaffold was hard, without enough elasticity following freeze-dry. The composite scaffold of 5: 5, 4: 6, 3: 7 and 2: 8 following freeze-dry was loose and soft, similar to sponge. With increased chitosan concentration,scaffold hardness increased. There were evenly distributed, detailed eyelets on the scaffold. Under the optical microscope,various pores were irregular; each pore closely connected and linked together; pore size was even, 20-100 μm. With increased chitosan concentration, pore size was gradually reduced. Scaffold porosity determination results displayed that mass ratio of silk fibroin/chitosan 4: 6 group > 5: 5 group > 3: 7 group > 2: 8 group. Compared with 2: 8 group, the porosity was significantly increased in the 5: 5 and 4: 6 groups (P < 0.05). No significant difference was detected in volume expansibility in the silk fibroin/chitosan composite scaffold of various mass ratios (P > 0.05). The degradation was slowest in the 2: 8 group, and fastest in the 5: 5 group at 4 weeks.CONCLUSION: Regarding physical and chemical properties, composite scaffold made by silk fibroin/chitosan showed significant superiority compared with scaffold made by silk fibroin or chitosan alone. Of them, silk fibroin/chitosan mass ratio of 5: 5 and 4: 6 are accorded with the requirement of cartilage tissue engineering.

Objective To discuss and evaluate the value of insertion of inferior vena cava filter in treating lower extremity deep venous thrombosis(DVT).Methods Inferior vena cava filters were placed in 46 patients with lower extremity DVT prior treatment,20 in which were treated by therapy with anticoagulation and thrombolysis,and therapy with pressure gradient,and the other 26 patients by operation and thrombolysis therapy,and therapy with pressure gradient.Whether patients occurred pulmonary embolism was observed and the form and site of filters were monitored by periodic fluoroscopy.Results Inferior vena cava filters were placed successfully in all patients,38 cases were implanted permanence inferior vena cava filter,8 cases were implanted temporary inferior vena cava filter.Symptoms and signs of DVT disappeared or remitted in 44/46 patients after treatment.None of pulmonary embolism was occurred.Follow up 2-24 months(average 13 months) for 36 cases with permanence inferior vena cava filter,there was no complication of the filter and pulmonary embolism occurred.Conclusions The method of inserting inferior vena cava filter is simple and safe,which can prevent pulmonary embolism effectually to offer sufficient safeguard for the treatment of DVT.

Objective: To investigate the contamination of phthalic acid esters (PAEs) and assess the health risk of PAEs contamination in market-available yellow rice wine in Huzhou City, Zhejiang Province, so as to provide the safety safeguard for consuming yellow rice wine. Methods: Yellow rice wine samples were collected from markets in Huzhou City from 2021 to 2022, and 16 PAEs were determined in yellow rice wine using magnetic solid-phase extraction coupled with gas chromatography-tandem mass spectrometry. The carcinogenic and non-carcinogenic risks of PAEs were evaluated using the health risk models proposed by United States Environmental Protection Agency. Results: A total of 75 yellow rice wine samples were collected, and 44 samples were detected with PAEs contamination (58.67%). Dimethyl phthalate (DMP), di-isobutyl phthalate (DIBP) and di-butyl phthalate (DBP) were detected, and there were 17 samples (22.67%) detected with DBP overdose (DMP and DIBP had no limit standard). DMP, DBP and DIBP, which were not classified as Class 2B and higher carcinogens by WHO's International Agency for Research on Cancer, had no definitive carcinogenic risks. Under mean PAEs, the five types of yellow rice wine all had no carcinogenic risks. Under 75% percentile of PAEs concentrations, the DBP in beverage wine with plastic packaging had a carcinogenic risk score of 1.207 5, with a gross carcinogenic risk score of 1.207 5. Under the maximum PAEs concentration, the ross carcinogenic risk scores of cooking wine with plastic packaging, beverage wine with plastic packaging, beverage wine with glass bottle packaging, and beverage wine with jar packaging were 2.751 0, 2.782 0, 1.298 2 and 2.944 0, presenting non-carcinogenic risks. Conclusion There is PAEs contamination in market-available yellow rice wine in Huzhou City, and no carcinogenic risk is evaluated. Non-carcinogenic health risk requires to be given a high priority.

Objective: To develop a magnetic solid-phase extraction combined with gas chromatography-tandem mass spectrometry (GC-MS/MS) based on pyrrole-modified magnetic nanoparticles to determine 16 types of phthalic acid esters (PAEs) in commercial liquors. Methods : Fe3O4 magnetic nuclei were prepared by hydrothermal synthesis, and the polypyrrole-modified magnetic nanomaterials were prepared with the chemical oxidation method. Magnetic solid-phase extraction of beer, grape wine, rice wine and Chinese spirits was performed at 10% alcohol by volume, extraction duration of 20 min and ethyl acetate elution of 10 min, followed by addition of 1 g NaCl for reduction of emulsification effect. The 16 types of PAEs were determined using GC-MS/MS with DB-5MS capillary column (30 m×250 μm, 0.25 μm) under the mode of electron impact ionization (EI) and dynamic multiple reaction monitoring (dMRM), with quantitative analysis using the external standard method. The standard curve, detection limit, spike recovery rate and precision of GC-MS/MS for determination of 16 types of PAEs were evaluated. Results: Pyrrole was successfully embedded onto the surface of magnetic nanoparticles in the form of polymer, and the magnetic nanoparticles modified by polypyrrole were well characterized, showing unapparent matrix and emulsification effects. There was a good linear relationship for the 16 types of PAEs at 50 to 5 000 ng/mL (r=0.999 5-0.999 9), and the spike recovery rate of 16 types of PAEs ranged from 71.61% to 110.50% at 100, 500 and 1 000 μg/kg, with relative standard deviations of 3.78% to 7.41%, detection limits of 0.02 to 1.47 μg/kg. PAEs were detected in 20 out of 50 liquor samples, with 30.00%, 60.00%, 40.00% and 70.00% detection rates in beer, grape wine, rice wine and Chinese spirits, respectively. Conclusions This method is sensitive to determine 16 types of PAEs in liquor samples, with unapparent matrix and emulsification effects, and the polypyrrole-modified magnetic composite nanoparticles present high adsorption of PAEs in liquor samples, which is feasible for monitoring of PAEs in multiple types of liquor samples.

Objective To evaluate short-term curative effect of endovascular abdominal aortic aneurysm repair. Methods Twelve cases of infra-renal abdominal aortic aneurysms were checked. Results The av-erage bleeding in the operation was 245 millimeter, the avenage hospitalization time was 8.6 days, and the average abrosia time was 1.5 days. White blood cell, hemoglobin, thrombocyte, hepatic function, and renal function were in the normal limits. Prothrombin time and activated partial thromboplastin time were post-poned after operation, and recovered normally within one week. Complications of operations were as follows:1 case of pulmonary infection, 2 cases of abdominal distention, and 1 case of intraoperafive endoleak. The former two kinds of complications alleviated after conventional treatment, and the latter disappeared naturally after 3 months. Conclusion Endovascular abdominal aortic aneurysm repair is safe, mini-invasive, and has little disturbance for body internal environment.

Objective To study the changes of anorectal motility and rectal sensation in the elderly patients with ulcerative colitis (UC). Methods The anorectal motility and rectal sensation were investigated by Medtronic PC-Polygraf HR made by Sweden in 35 non-elderly patients versus 19 elderly patients with UC, and 20 non-elderly healthy subjects (HS) and 28 elderly HS were as control group. Results (1) The static pressure, pressure of anal sphincter and the maximal squeeze pressure of anal sphincter in non-elderly patients and elderly patients with UC showed no significant differences compared with those in non-elderly HS and elderly HS group (elderly patients with uc vs. ederly HA:t= 1.311,1.298,1.401;nonederly patients with uc vs. nonederly HS: t=1.294,1.299,1.322all P＞0.05). When abdominal pressure was increased, the net increased pressure of anal sphincter was (2.8±1.1) kPa in the elderly patients with UC, (2.9±1.3) kPa in the non-elderly patients with UC. The pressures were lower in two UC groups than in HS groups [elderly HS group:(3.8±1.2) kPa; non-elderly HS group:(3.9±1.2) kPa,elderly patients with uc vs. ederly HS:t=2.238,nonelderly patients with us vs. nonederly HS:t=2.243 all P＜0.05]. (2)The rectal lowest volume of sensory threshold, the maximal volume of tolerance and the maximal compliance were (85±30) ml, (180±69) ml, (26. 5±8.8) ml/kpa in elderly patients with UC and (65±15) ml, (170±58) ml, (22.6± 10. 3) ml/kPa in non-elderly patients with UC. They were lower than in each HS group [elderly HS group (95±31) ml, (205±78) ml, (32.9±12.9) ml/kPa; non-elderly HS group:(78±38) ml, (190±50) ml, (30.8± 15.2) ml/kpa, all P＜0. 01]. (3)The rectal lowest volume of sensory threshold, the maximal volume of tolerance and the maximal compliance in elderly patients with UC were higher than in non-elderly patients with UC (elderly patients with uc vs. elderly HS:t=3. 121,3. 135,3.146,nonederly patients with uc vs. non elderly HS: t= 3.162, 3.141, 3.188 elderly patients with uc vs. nonelderly patients with uc: t = 2. 246,2. 239,2. 240 all P＜ 0. 05). The rectal lowest volume of sensory threshold, the maximal volume of tolerance in elderly HS group were higher than in non-elderly HS group (ederly HS vs. t = 2. 328,2. 301 all P＜0. 05). Conclusions There are some anorectal motility disturbances in UC. Higher sensitivity, lower tolerance, lower compliance of rectum and weakened anal automatic control function in UC may be associated with diarrhea and frequent defecation. The rectal sensation threshold to volume stimulus is higher in elderly HS than in non-elderly HS group. The sensibility to volume ectasis of rectum is weakened and the survivability of rectum is increased in elderly patients with UC.

Background and purpose:It was reported that erythropoietin may directly or indirectly induce the tumor cells to proliferate and result in diseases progression when recombinant human erythropoictin is used clinically in cancer-related anemia.Recently,the expression of erythropoietin-receptor(Epo-R)was detected in breast cancer.This study was done to detect the expression of Epo-R,estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor 2(Her-2)in breast cancer,and to investigate the relationships between these indexes and the clinical significance.Methods:Sixty breast cancer patients were analyzed,the expression of Epo-R and microvascular density(MVD)were detected by immunohistochernistry,in order to clarify the relationships between the expression level of Epo-R,MVD and ER,PR,Her-2.Results:The rate of Epo-R expression was 78.3%,the mean number of positive tumor cells was 39±24,while the tissue MVD was 25±9.The expression level of Epo-R was positively correlated with the MVD.Also,the level of MVD was higher in the group of Epo-R positive than the negative,which has significant difference(t=3.4252,P＜0.001).The expression level of ER,PR has no definite relationships with Epo-R,MVD.The expression level of Her-2 was both closely associated with Epo-R,MVD.The expression level of Epo-R has an obvious relationship with clinical stage,lymph node status and the size(P＜0.001).The value of MVD was positively correlated with lymph node status,while not with clinical stage and the size.Conclusion:The expression level of Epo-R was markedly higher in breast cancer,and has a positive correlation with the tissue MVD.The expression level of Epo-R and MVD were significantly associated with Her-2,but not with ER and PR.It may contribute to clarifying the clinicopathological characteristics and prognosis of breast cancer by detecting both the expression level of Epo-R and MVD.

Objective To investigate the diagnostic and prognostic value of neutrophil-to-lymphocyte rate ( NLR) in acute pancreatitis related acute kidney injury patients .Methods Peripheral blood specimens and clinical information of 98 acute pancreatitis patients in Wenzhou Center Hospital were collected .The WBC, neutrophils , lymphocytes were detected and NLR were calculated when they were admitted .The patients were divided into two groups by their NLR and whether they were with AKI respectively .Besides, ALT, Hct, TC, TG, blood calcium concentration , serum creatine and urea nitrogen , C reactive protein were detected and patients′APACHEⅡ score were also recorded to analyze the difference between the two groups .Results There is no significance in the age , BMI, Hct, TG, TC, ALT and blood calcium between AP patients with and without AKI.The blood creatinine, BUN, CRP, APACHEⅡscore were (395 ±122)μmol/L, (28.2 ±5.2) mmol/L, (34.0 ±8.2)mg/L, (11.5 ±3.8) score, respectively in AP patients with AKI, and which were (79 ±17 )μmol/L, ( 7.3 ±2.0 ) mmol/L, ( 14.8 ±2.9 ) mg/L, ( 6.9 ±2.4 ) score, respectively in AP patients without AKI.The blood ALT, blood creatinine, BUN, CRP, APACHEⅡscore were (257 ±76)U/L, (159 ±62)μmol/L, (20.5 ±6.6)mmol/L, (24.8 ±5.5)mg/L and (12.4 ±4.6) score in the patients with higher NLR respectively , and which were ( 165 ±30 ) U/L, ( 98 ±23 )μmol/L, ( 14.3 ±5.2 ) mmol/L, (19.5 ±3.0)mg/L and (5.4 ±2.1) score in the patients with lower NLR respectively .NLR was 4.97 ±0.19 in AP patients with AKI, and was 9.62 ±0.81 in AP patients without AKI.The difference between the two groups was significant(P=0.0001).The area under ROC curve of diagnosing AP by NLR was 0.895 (95%CI 5.75).the sensitivity was 89.5%and the specificity was 77.2% when using 5.75 as the cut-off value to diagnose AP related AKI with NLR .Conclusions NLR can be a potential predictive index of the severity and relate to renal function in acute pancreatitis related acute kidney injury patients .

Objective To assess the curative effect of percutem transilluminated with negative pressured on the potaried technique on the treatment of venous ulcer in lower extremity.Methods The clinical date of 300 cases involving 300 legs with venous ulcer in lower extremity,who underwent the percutum transilluminated negative pressured potaried technique using TRIVEXTM Ⅱ potaried system or the percutum transfixion surgical treatment from October 2005 to June 2009,were analyzed.Three hundred cases were randomly divided into potaried group and transfixion group.In potaried group,there were 190 cases involving 190 legs treated with TRIVEXTM Ⅱ potaried system.In transfixion group,110 cases involving 110 legs treated with percutum transfixion.The clinical indexes of skin infection rate and skin necrosis rate,shrinkage rate of wound area and skin depigmentation rate,ulcer healing rate and ulcer recurrence rate were calculated to assess the clinical curative effect on day 5,day 20,day 120 and day 360 after operation respectively.Results The rates of skin infection and skin necrosis were significantly decreased in potaried group compared with transfixion group on day 5 after operation (P0.05).Ulcer recurrence rate was remarkably lower in potaried group than that in transfixion group on day 360 (P