BACKGROUND: Thermosensitive hydrogel materials present stability at human body temperature, which is necessary for its application as a medical implant, thus the lower critical solution temperature (LCST) of thermosensitive hydrogel should be beyond the human body temperature by adjustment.OBJECTIVE: To prepare a thermosensitive hydrogel poly-N-isopropylacrylamide-co-N-hydroxymethylacrylamide P(NIPAAm-co-NHMPA) with over 37 ℃ LCST, and primarily appraise its safety as a medical implant in vivo.DESIGN: Random, non-blind, group control, animal experimental study.SETTING: Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology(HUST).MATERIALS: The experiments were carried out in the Central Laboratory, Union Hospital, Tongji Medical College, HUST and the Key Laboratory of Biomedical Polymer Materials of the Ministry of Education, College of Chemistry, Wuhan University between January and October in 2007. NIPAAm monomer and NHMPA monomer were purchased from Aldrich Company, crosslinking agent N, N'-methylene bisacrylamide from Fluka Company, and initiator ammonium persulfate and accelerating agent tetramethyl ethylene diamine from Sigma Company.METHODS: ①Taking ammonium persulfate and tetramethyl ethylene diamine as oxidation-reduction initiation system, while N, N'-methylene bisacrylamide as cross-linking agent, P-NIPAAm-co-NHMPA was prepared with the addition of NHMPA in the reaction system. LCST was determined by shrinking tests.②A series of biocompatibility tests such as sensitization test, acute systemic toxicity test, genetic toxicity test and implantation test were conducted in several experimental animals to evaluate the safety of the implant. MAIN OUTCOME MEASURES: The erythema and edema of stimulated lesions were recorded in sensitization test; the general state of each animal in acute systemic toxicity test were recorded 4, 24, 48 and 72 hours after injection; in genetic toxicity test, mouse bone marrow polychromatic erythrocyte (PEC) micronucleus was counted six hours after injection under microscope; sections after implantation were observed under light microscope.RESULTS: ①Synthesized hydrogel showed thermosensitive character and the LCST was 38 ℃.②In sensitization test, there was no erythema and edema occurred after leaching liquor and saline were injected; acute systemic toxicity test result revealed no symptom of toxicity; the genetic toxicity test suggested no difference of PEC frequency between experimental group and negative control group; in vivo implantation test, the inflammation around the material was mild and limited.CONCLUSION:P(NIPAAm-co-NHMPA) shows good biocompatibility and can be potentially used as an implant material.

BACKGROUND: Caspase-3 is a cysteine proteinase that can promote cell apoptosis. Ciliary neurotrophic factor has many kinds of biological activities, such as protecting various neurons from injury, especially motorial neurons, thereby reducing the occurrence of nerve cell injury.OBJECTIVE: To observe the Influence of ciliary neurotrophic factor on spinal cord caspase-3 expression in different ages rats following sciatic nerve injury, aiming to make further investigation about the changing regularity and modulating character of peripheral nerve damage at various ages rats.DESIGN: Randomized controlled experiment.SETTING: Ultrasound Department and the 5th Research Institute of Daping Hospital, Field Surgery Research Institute of the 3rd Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out at the Field Surgery Research Institute of the 3rd Military Medical University of Chinese PLA from April 2000 to December 2001. Altogether 810 wistar rats including infant rats with body mass of 30-100 g (birth age of 20 days), adult rats of 200-350 g (birth age of 4 m), elder rats of 400-800 g (birth age of 18-24 m),were selected with 270 rats in each age stage. Female and male does not limit.METHODS: [1] Experimental animal grouping: Various age rats were randomly divided into normal group (n=18), ciliary neurotrophic factor group(n=126) and physiological saline group (n=126), rats in ciliary neurotrophic factor group and physiological saline group were subdivided into 1 day, 3days, 1 week, 2 weeks, 4 weeks, 8 weeks, 12 weeks subgroups. [2] Animal model preparation: In Ciliary neurotrophic factor group and physiological saline group, rats were cut off a piece of sciatic nerve of 2 mm long, both ends connected with silica tube for constructing neuranagenesis cab, in which 15 μL recombined ciliary neurotrophic factor (25 mg/L) was injected in ciliary neurotrophic factor group, and 15 μL physiological saline in physiological saline group. [3] Preparation and examination of specimen:Six rats were randomly selected from each group, lumbar L3-5 spinal cord were obtained for carryingonnation, caspase-3 activity examination and Western blotting examination.MAIN OUTCOME MEASURES: [1] Expression of spinal cord caspase-3 with IHC assay. [2] Distribution and expression intensity of spinal cordcaspase-3 by Western blotting technique. [3] Changes in caspase-3 activity.RESULTS: Altogether 810 rats completed the experiment, all data was entered the final result analysis. [1] Expression of spinal cord caspase-3 with IHC assay: After injury, neuronal caspase-3 expression became strengthened at injured side of various age physiological saline groups, which obviously increased at posttraumatic 2 weeks and 4 weeks, but less expressed at 8 weeks and 12 weeks; while in ciliary neurotrophic factor group, posttraumatic caspase-3 expression was mostly obvious at 2 weeks and 4 weeks. [2] Distribution and expression intensity of spinal cordcaspase-3 by Western blotting technique: Caspase-3 expression was not significant difference between various age subgroups in normal group(P ＞ 0.05). Comparing to the same age normal group and non traumatic group, caspase-3 expression was strengthened at 1 week, 2 weeks, 4 weeks in various age physiological saline group and ciliary neurotrophic factor group damages (P ＜ 0.05-0.01); in ciliary neurotrophic factor group, caspase-3 showed weaker expression at 1 week, 2 weeks, 4 weeks in infants; 2weeks and 4 weeks in adults, 4 weeks in elders comparing to corresponding physiological saline group, difference was significant (P ＜ 0.05-0.01).The comparison between untraumatic side of each group and normal group showed no statistical difference (P ＞ 0.05). [3] Changes in caspase-3 activity: In child, adult and elder physiological saline groups, caspase-3 activity was increased in traumatic spinal cord, moreover caspase-3 activity was higher than elder and adult group at various age point (P ＜ 0.05-0.01); in child, adult and elder ciliary neurotrophic factor groups, caspase-3 activity is lower than physiological saline group (P ＜ 0.05-0.01). After damage,caspase-3 activity at traumatic side in physiological saline group and ciliary neurotrophic factor group was difference from normal group but without significant meaning (P ＞ 0.05), except the expression in child posttraumatic 12 weeks group was lower than normal group (P ＜ 0.05).CONCLUSION: After sciatic nerve damage, caspase-3 protein expression and activity were found to be increased in spinal cord of different age groups rats which can be reduced by extragenous ciliary neurotrophic factor, which playing protective role on spinal cord nerve cells, such protection would gradually attenuated in child group, adult group to elder group in turns.

AIM: To prepare and evaluate the biocompatibility of a thermosensitive hydrogel poly-(N-isopropylacrylamide-co-N-hydroxymethylacrylamide) (P-NIPAAm-co-NHMPA) in vivo so as to appraise its safety as a medical implant in clinical use. METHODS: The experiments were carried out in the Central Laboratory, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology(HUST) and the Key Laboratory of Biomedical Polymer Materials, Ministry of Education, College of Chemistry, Wuhan University between January and October in 2007. ① Preparation of thermosensitive hydrogel P-NIPAAm-co-NHMPA: Taking ammonium persulfate and tetramethyl ethylene diamine as oxidation-reduction initiation system, while N, N'-methylene bisacrylamide as cross-linking agent, P-NIPAAm-co-NHMPA was prepared with the addition of NHMPA in reaction system. Low critical solution temperature was determined by shrinking tests. ②A series of biocompatibility tests such as sensitization test, acute systemic toxicity test, genetic toxicity test and implantation test were conducted to evaluate the safety of the implant. RESULTS: ①Synthesized hydrogel showed thermosensitive character as expected, and the low critical solution temperature was 38 ℃.②In sensitization test, there was no sensitization proved in the material; in acute systemic toxicity test, the symptom of toxicity was not found; the genetic toxicity test suggested no teratogenic or mutagenic effects; in vivo implantation test showed that the inflammation around the material was mild and limited. CONCLUSION: Thermosensitive hydrogel P-NIPAAm-co-NHMPA has good biocompatibility and is a potential medical implant.

BACKGROUND: Thermosensitive hydrogel has made great progresses in drug carrier and tissue engineering. However, its phase-transition temperature is lower than human body temperature, so it has not been used as implants. OBJECTIVE: To prepare and evaluate the biocompatibility of a thermosensitive hydrogel poly-(N-isopropylacrylamide-co-N-hydroxymethylacrylamide)[P(NIPAAm-co-NHMPA)] In vitro so as to assess its safety as a medical implant.DESIGN, TIME AND SETTING: Material experiment was performed at the Central Laboratory of Union Hospital of Tongji Medical College, Huazhong University of Science & Technology and Key Laboratory of Biomedical Polymers, Ministry of Education, Wuhan University Chemical Department from January to October 2007.MATERIALS: L929 mice fibroblast line was provided by Professor Chen, Stomatology Hospital of Wuhan University. N-isopropylacrylamide and N-hydroxymethylacrylamide were purchased from Aldrich.METHODS: Using ammonium persulfate and tetramethylethylenediamine as oxidateion-reduction initiation system and N, N'-methylene bisacrylamide as crosslinking agent, mass fraction 8.5% N-hydroxymethylacrylamide was added to the reaction system. The N-isopropylacrylamide and N-hydroxymethylacrylamide were dissolved in deionized water and reacted with N, N'-methylene bisacrylamide, ammonium persulfate and tetramethylethylenediamine at room temperature for 3 hours. The colorless transparent cylinder gel was obtained, soaked in deionized water and dried.MAIN OUTCOME MEASURES: Biocompatibility in vitro of the material was validated by cytotoxicity test, cells/scaffolds interaction test, hemolysis test and pyrogen test in vitro.RESULTS: Synthesized hydrogel showed thermosensitive character as expected and the lower critical solution temperature was 38 ℃. The toxicity rate was grade 0-1. Through the scanning electron microscopy, cells and scaffolds were tightly attached, producing a perfect interface, and the shape of cells was all right. Hemolysis rate was 2.54%. No pyrogen reaction was found. CONCLUSION: P(NIPAAm-co-NHMPA) has been successfully prepared with good biocompatibility in vitro and might become a good medical implant for clinical use.

Objective To summarize the clinical and pathological characteristics of primary gallbladder carcinoma, analyze factors such as surgical procedures, tumor stages that associated with prognosis, and standardize the diagnosis and treatment strategy for primary gallbladder carcinoma.Methods From January 1986 to October 2009, 197 patients of primary gallbladder carcinoma were treated in Peking Union Medical College Hospital (PUMCH). A retrospective study was performed basing on clinical data of these patients, statistical analysis carried out using SPSS version 13.0, Kaplan-Meier estimate employed for the survival analysis and Log-rank test for the group comparisons. Results One hundred and ninety-seven patients were enrolled (73 male and 124 female) with the mean age of 64.4±1.1years. Abdominal pain (77.3%) , obstructive jaundice (33.6%) and fever (19.5%) were the main symptoms of primary gallbladder carcinoma, with accompanying cholecystolithiasis in 99 cases. B-mode ultrasonic examination and CT scan were important for preoperative diagnosis. Nevin stages and surgical modalities were independent prognosis factors. For stage Ⅰ and Ⅱ patients the survival terms were not different between those receiving radical cholecystectomy and simple cholecystectomy. However, for stage Ⅲ, Ⅳ and Ⅴ patients the survival terms were of statistically different between those in radical cholecystectomy group and simple cholecystectomy group. Conclusions For stage Ⅰ and Ⅱ patients, simple cholecystectomy is a good surgical procedure. For those at more advanced stages, radical cholecystectomy is the choice of therapy, while simple cholecystectomy also improves the prognosis compared with laparotomy only.

AIM:To investigate the changes of autophagy of macrophages after phagocytizing dust particles and explore the mechanisms of dust particle-induced autophagy of the cells.METHODS:The bronchopulmonary lymph nodes were collected from patients with lung operation.The paraffin sections were prepared and then stained with Wilder's method.The tissue structures were viewed.Peritoneal macrophages were harvested from rats and then treated with carbon particles.Influences of carbon particles in autophagic activities of macrophages were examined.The ultrathin sections of the lymph nodes and the cells phagocytized carbon particles were prepared.The structures and distribution of phagosomes,autophagosomes and lysosomes were viewed.The apoptotic cells in the dust cells of the lymph nodes and the cells having phagocytized carbon particles were examined using transmission electron microscope and TUNEL staining.RESULTS:In adult lymph nodes,dust particles were deposited significantly in macrophages,collagen fibres and density of microvessels increased.There were autophagosome precursors,autophagosomes,autophagolysosomes as well as phagosomes in the dust cells and the cells phagocytized carbon particles.In autophagosomes,mitochondrion,dust particle or carbon particle were usually observed.There were positive cells by TUNEL staining in the dust cells and the cells phagocytized carbon particles.Nuclear condensation or apoptotic body in the apoptotic cells were observed under transmission electron microscope.CONCLUSION:Deposition of dust particles induces enhancement of autophagic activities and apoptosis of macrophages.Autophagy plays an important role in cleaning dust particles and the injured mitochondria.

Objective To summarize the clinical and pathological characteristics of incidental gallbladder carcinoma,analyze the factors such as surgical procedures,tumor stages that associated with prognosis,and standardize the diagnosis and treatment strategy of incidental gallbladder carcinoma. Methods Between January 1991 and October 2009,27 incidental gallbladder carcinoma patients were treated in Peking Union Medical College Hospital (PUMCH).A retrospective study was performed based on the clinical data of these patients to evaluate the methods of diagnosis,curative treatment and prognosis. Results Twenty-seven patients were collected (8 male and 19 female) with the mean age of (68.3 ± 1.9)years.Abdominal pain,obstructive jaundice and fever were the main symptoms of incidental gallbladder carcinoma.All the 27 cases underwent open cholecystectomy based on preoperative tentative diagnosis of benign disease,and carcinoma of gallbladder was confirmed by postoperative pathology.In this group(9 lowdifferentiation,9 moderate-differentiation,4 high-differentiation and 5-carcinogenesis from adenoma).Two cases were on the stage of Nevin Ⅰ,5 on Nevin Ⅱ,8 on Nevin Ⅲ,5 on Nevin Ⅳ and 7 on Nevin Ⅴ.The survival terms were of statistic significance between the radical cholecystectomy groups and simple cholecystectomy groups(x2 =4.450,P=0.035); The prognosis of patients on stages Ⅰ、Ⅱ are better than those who are on stages Ⅲ、Ⅳ、Ⅴ (x2 = 6.825,P = 0.014). Conclusion The clinical manifestations of incidental gallbladder carcinoma are diverse,which usually lead to misdiagnosis.Frozen section examination during operation is key to diagnose incidental gallbladder carcinoma.Surgical operation is the first choice of therapy,offering much better prognosis than those received simple cholecystectomy.

Objective To explore risk factors for type 2 diabetes mellitus complicated with gallstone disease. Methods The clinical data of 429 type 2 diabetes mellitus patients complicated with gallstone disease (case group) and 2145 type 2 diabetes mellitus patients without gallstone disease (control group) were collected from the Health Center of Peking Union Medical College Hospital from January 2007 to May 2010. According to sure (SBP), diastolic blood pressnre (DBP), fasting blood glucose (FBG), total cholesterol (TC), Triglyceride (TG), high-density lipoprotein cholesterol (HDL-CH), low-density lipoprotein cholesterol (LDL-CH) and body mass index (BMI) were statistically analyzed by fourfold table chi-square test or conditional Logistic regression.Results Univariate analysis showed that there was a significant difference in the levels of SBP, FBG, TC, HDL-CH and BMI between the case group and the control group (x2 =20.323, 4.365, 4.028, 7.049, 7.319, P＜0.05). Multivariate analysis confirmed that SBP, TC, HDL-CH and BMI were risk factors for gallstone disease in patients with type 2 diabetes mellitus (x2 = 18. 047, 6. 905, 12. 884, 7. 557, P ＜ 0. 05). Conclusion Increased SBP, TC and BMI and decreased HDL-CH may be the risk factors for type 2 diabetes mellitus complicated with gallstone disease.

Objective To investigate the risk factors of gallbladder carcinoma,so as to provide theoretical base for the prevention of gallbladder carcinoma.Methods The clinical data of 153 patients with gallbladder carcinoma (gallbladder carcinoma group) who were admitted to the Peking Union Medical College Hospital from January 2000 to December 2010 were retrospectively analyzed. A total of 300 patients with cholecystolithiasis (cholecystolithiasis group) and 300 patients without gallbladder carcinoma or cholecystolithiasis (control group)were collected and matched at the ratio of 1∶2 to conduct the controlled study.Data were statistically analyzed by the Chi-square test and conditional Logistric regression.Results Univariate analysis showed significant difference in age,history of cholecystolithiasis,postmenopausal age,accumulated menstrual period,giving birth or not and number of birth between gallbladder carcinoma group and control group ( x2 =58.22,180.14,9.59,24.30,18.66,15.17,P ＜0.05).Age,history of cholecystolithiasis,accumulated menstrual period and number of birth were the independent risk factors of gallbladder carcinoma (x2 =55.76,180.95,24.30,8.54,P ＜ 0.05).The risk of having gallbladder carcinoma in patients who had a history of cholecystolithiasis was 34 times higher than those who did not have the history of cholecystolithiasis (OR =34.22).Late postmenopausal age (51 -55 years old),longer accumulated menstrual period ( ≥30 years),and the number of birth ( 3 times) were associated with higher risk of gallbladder carcinoma (OR =3.96,9.68,3.51 ). Age,course of cholecystolithiasis and accumulated menstrual period and number of birth were the risk factors of gallbladder carcinoma when comparing patients who have history of cholecystolithiasis in the gallbladder carcinoma group with those in the cholecystolithiasis group (x2 =70.66,16.66,11.59,4.69,P ＜ 0.05 ).Age,course of cholecystolithiasis and accumulated menstrual period were the independent risk factors of gallbladder carcinoma ( x2 =64.29,8.82,5.58,P ＜ 0.05).The risk of gallbladder carcinoma increased as the increase of age and course of cholecystolithiasis. The accumulated menstrual period ≥ 30 years was also a risk factor of gallbladder carcinoma. Conclusions Age,history of cholecystolithiasis,course of cholecystolithiasis,accumulated menstrual period and number of birth may be the risk factors of gallbladder carcinoma.For patients with age above 60 years and course of cholecystolithiasis above 3 years,cholecystectomy should be conducted to reduce the incidence of gallbladder carcinoma,and great importance should be attached to female patients with indications mentioned above.

Insulin resistance, lipid metabolism disorders, independent autonomic neuropathy are high risk factors for gallstone formation in type 2 diabetic patients. In recent years, there have been a lot of new developments on the relationship between type 2 diabetes and gallstones, such as adiponectin, leptin and metabolic syndrome, etc. Based on these studies, the risk factors of type 2 diabetes complicated with gallstones are summarized in this paper.