Objective To prepare the small intestinal submucosa (SIS) to be used as a scaffold in constructing engineered cardiac tissue. Methods Small intestinal submucosa of pig was treated with 0.25% trypsin and then 0.5% SDS for 24 hours each to obtain decellularized SIS. The obtained SIS was tested for its mechanical strength by stretching both horizontally and longitudinally with a material test machine. The cytotoxicity and histocompatibility of the material was also examined. Then the cardiomyocytes harvested from 3-day old neonatal rats were seeded on the SIS to construct engineered cardiac tissue sheets. These acquired engineered cardiac tissue sheets were immunohistochemically stained and observed with inversion microscope and transmission electromicroscope to evaluate their characteristics. Results SIS was decellularized completely. The mechanical capability of the decellularized SIS was satisfactory. Under 15% stretching, its strain and stress was nearly linear. SIS showed no cytotoxicity and inflammatory response. After 12 hours, the cardiomyocytes seeded in the SIS scaffold began to beat spontaneously. Two days later, the cardiomyocytes-SIS scaffold composite (engineered cardiac tissue sheet) began to beat spontaneously, and could beat spontaneously for 14 days. The constructed engineered cardiac tissue sheets consisted of layers of cardiomyocytes and with abundant extracellular collagen in the sheet. Conclusions SIS with good mechanical capability and biocompatibility has been prepared successfully, and then the engineered cardiac tissue has been constructed successfully based on the SIS scaffold.

Objective To study the effects of midazolam on expression of vascular endothelial growth factor (VEGF) of gerbils following total cerebral ischemia-reperfusion injury to look for an experimental basis for the rational clinical use of midazolam. Methods Seventy-two male gerbils (Mongolian gerbil) were randomly assigned into three groups (24 each): sham injury group, injury group and midazolam treatment group. Total cerebral ischemia was reproduced by blocking the bilateral carotid arteries for 10 minutes with bulldog clamps. When reperfusion began, with release of the clamps, 5mg/kg of midazolam was intraperitoneally injected to the animals in midazolam group, and 50ml/kg of normal saline was given by the same way in the gerbils in injury group. Then the parameters listed below were observed: positron emission tomography (PET) images at 6h, 1d, 3d and 7d after reperfusion, and the expression of VEGF in cerebral tissue was immunohistochemically assessed. Results No obvious abnormality was found in the cerebral tissue of sham injury group. For the animals in the injury group and midazolam treatment group, the brain reinfusion area enlarged obviously (P

Objective To analyze the immune state and discuss the effects of immunoregulation therapy of combined use of ulinastatin and thymosin ?1 in the treatment of posttraumatic sepsis. Methods Forty-eight patients with posttraumatic sepsis admitted to ICU during Oct. 2005 to Oct. 2007 were included and randomly divided into treatment group and control group (24 each). Patients in control group received conventional SSC treatment, while those in treatment group received conventional SSC treatment plus immunoregulation therapy with a combination of ulinastatin and thymosin ?1. Peripheral blood CD14+ monocyte HLA-DR expression was detected by flow cytometry to determine the innate immunity of posttraumatic septic patients on day 1, 5, 7 and 28 (or death), and serum levels of IL-6 and IL-10 were assessed by ELISA to evaluate the nonspecific immunity at day 1 and 28 (or death). APACHE Ⅱ scores were recorded on day 1, 5, 7 and 28 (or dearth). Ten healthy volunteers were enrolled as healthy controls. Results At admission the expression of CD14+ monocyte HLA-DR was lower than 30% in 9 posttraumatic septic patients. The expression levels of CD14+ monocyte HLA-DR in the patients in treatment group were much higher than those in control group (P0.05). Before treatment, no significant difference existed in APACHE Ⅱ scores between treatment group and control group, while this score in the patients in treatment group was much lower than that in control group on day 5, 7 and 28 (P0.05). Conclusions There is immunosuppression in some posttraumatic patients with somplication of sepsis. All these patients manifest exaggerated nonspecific immunity at the early stage, and then manifest immunodepression as sepsis progresses. The immunoregulation effects of a combination of ulinastatin and thymosin may facilitate the balance of pro-inflammatory and anti-inflammatory media, thus ameliorating septic symptoms. No improvement in 28 day mortality has been seen, probably due to limited number of patients in present study.

Objective To investigate and analyze the state of injury and treatment for the patients with maxillofacial injuries in Wenchuan earthquake, and search for more effective methods for treating maxillofacial injury. Methods One hundred and sixty-five patients wounded in Wenchuan earthquake were admitted to authors' hospital, and they received specialized diagnosis and treatment. In this paper, the emphasis was put on those patients with severe ear-nose-throat and maxillofacial injuries. Based on the analysis of the state of injuries, the patients were treated with debridement and suture, reshaping and suture, reduction and fixation of fracture, or local or systematic anti-infection therapy. All the treated patients were followed up 2 weeks after treatment. Results Thirty-three of the 165 admitted patients suffered from various degrees of maxillofacial injury (20.0%), including 10 cases of ear injuries (6.1%), 8 cases of nasal trauma (4.8%) and 2 cases of lip laceration (1.2%). All the wounds healed by primary intention after the treatment with no disfiguration. 8 cases of nasal sinus and zygomatic fractures (4.8%) and 5 cases of mandibular fracture (3.0%) recovered nicely after the treatment. No trauma was found in throat, neck, trachea and esophagus. 6 patients (3.6%) were complicated with infection after injury (onset of infection outside of hospital). There was no death. Conclusion Patients with maxillofacial injury should receive early and appropriate treatment. Besides restoration of function, it is also important to restore the physiognomy

Objective To investigate the effect of mytomycin C in the operation of dacryocystorhinostomy for curing lacrimal passage obstruction. Method Forty-one patients with traumatic lacrimal passage obstruction were divided into treatment group (n=21, 14 males and 7 females with an average age of 32) and control group. (n=20, 12 males and 8 females with an average age of 35). The patients in treatment group received 0.4mg/ml of mytomycin C when undergoing dacryocystorhinostomy, while the patients in control group received no medicine when undergoing the same operation. Lacrimal passage was cleaned with water after operation, and the operational effectiveness of dacryocystorhinostomy were compared between the treatment group and control group. Results Twenty-one eyes were cured in treatment group, and the cure rate reached 100%. While for the 20 eyes in control group, 17 eyes were cured, and the cure rate was 85%. There existed statistical significance between the two groups (P

Objective To summarize the experience of examination and treatment of 237 cases of eye diseases with portable slit lamp in earthquake area of Dujiangyan, Sichuan Province, from May 14 to July 8, 2008. Methods 237 cases of the eye diseases were examined and diagnosed. With portable slit lamp, the foreign bodies in 25 cases of conjunctival foreign body and 62 cases of corneal foreign body were removed. Results Of the 237 cases, 39 were diagnosed as conjunctivitis, 25 as conjunctival foreign body, 13 as pterygium, 27 as keratitis, 11 as corneal ulcer, 62 as cornea foreign body, 24 as cornea injury, 7 as hyphema, 16 as iridocyclitis, 5 as glaucoma and 8 as cataract. With portable slit lamp, the foreign bodies in 25 cases of conjunctival foreign body and 62 cases of corneal foreign body were removed in one lump with no corneal perforation or infection. Follow-up examination showed that the wounded cornea repaired pretty well. Conclusions With portable slit lamp, ophthalmologists can not only make a precise diagnosis for the eye diseases in the anterior segment, but also are able to timely to remove foreign bodies in conjunctiva and cornea. Portable slit lamp is a useful and convenient instrument for ophthalmologists in the field of earthquake disaster relief.

Objective To screen the tumor specific antigens, and explore new methods for diagnosis and therapy of tumor. Methods Hela cells, the human cervical carcinoma cell line, were used to immunize the BALB/C mice, and the positive hybridomas were screened by rosette formation screening system in human carcinoma cell line Colo205, Burkitt lymphoma cell line, Raji, etc. The immunofluorescence staining and flow cytometry analysis were performed to characterize if the prepared monoclonal antibodies (mAb) could reorganize the natural membrane antigen expressed on Hela, Colo205, Raji, PLA801, 7901, Jurkat, 293T, ECV304 and resting or activated human peripheral blood mononuclear cells (PBMC). Furthermore, the membrane protein of Hela, Raji and Sp2/0 cells was segregated by ultracentrifugation and the mAbs were cross-linked with the Sepharose CL-4B. In order to identify the molecules which could be recognized by the prepared mAbs, immunoprecipitation was performed to purify the tumor antigen candidates with the cell membrane protein from Hela, Raji and SP2/0 (as negative control) through Sepharose CL-4B cross-linked with the prepared mAbs. Finally, western blotting was performed to detect the molecular weight of the products from the immunoprecipitation. Results Three mAbs were obtained which could recognize the tumor antigen candidates expressed in several human carcinoma cell lines, such as Hela, Raji and Colo205, and some embryonic cell lines, such as 293T and ECV304, but they could not bind to Jurkat, PLA801, 7901, SP2/0, resting or activated human PBMC. Western blotting results indicated that the three mAbs could recognize a 47kD molecule which expressed in Hela and Raji but not on SP2/0 cell membrane. Conclusion Three mAbs, which can recognize tumor membrane antigen candidates, are obtained, providing useful tools for screening and identify the tumor specific antigens.

Objective To investigate the relationship between the expression of metastasis associated antigen 1 (MTA1) and the biological behavior of endometrial carcinoma. Methods From May 2005 to May 2007, 30 patients with endometrial carcinoma and 20 patients with normal endometrial tissues, were admitted to Changzheng Hospital of the Second Military Medical University for surgical treatment were involved in the present study. The expression of MAT1 mRNA in two different groups was detected by real-time RT-PCR, and of MAT1 protein was detected by immunohistochemical streptavidin-peroxidase (SP) method and Western blotting. The relationship between their expression and pathological features were analyzed. Results The positive expression rate of MTA1 protein was 80% in endometrial carcinoma group, which was significantly higher than that in control group (30%, P0.05). Conclusion The expression of MTA1 might play a potential role in the pathogenesis and development of endometrial carcinoma, and it might be a novel target for the therapy and prognostic indicator of endometrial carcinoma.

Objective To study the expression of DNA methyltransferase 1 (DNMT1) in human cervical carcinoma tissue and cells, analyze the relationship between the expression level of DNMT1 and the clinicopathologic features of cervical carcinoma, and explore the role DNMT1 plays in tumorigenesis of cervical carcinoma. Methods The expression levels of DNMT1 protein in 3 cervical cancer cell lines, 89 samples of cervial cancer tissue, 20 samples of cervical intraepithelial neoplasia (CIN) and 20 samples of normal cervix were determined by immunohistochemistry method. The expression levels of DNMT1 mRNA in all the cell lines and 79 paraffin sections of cervix tissue were determined by RT-PCR. Results The DNMT1 protein was expressed in all the three cervix cancer cell lines. Positive expression was detected in 71.9% of cervix cancer samples, 35.0% of CIN samples and 10.0% of normal cervix samples, with significant differences among the three groups (P

Objective To study the effects of recombinant adenovirus vector containing vp3 gene (AdAFPvp3) on c57BL/6 murine hepatocellular carcinoma model. Methods The titers of adenovirus vectors AdAFPvp3 and AdCMV-eGFP were detected. c57BL/6 murine hepatocellular carcinoma models were reproduced by subcutaneous inoculation of murine Hepa1-6 cells, and tumors were monitored for their generation rates. Twenty-four mice were randomly divided into adenovirus vector AdAFPvp3 group, adenovirus vector AdCMV-eGFP control group and PBS control group (8 each) when the tumors grew into 5mm in diameter. AdAFPvp3 (5?108pfu/100?l), AdCMV-eGFP (5?108pfu/100?l) and PBS (100?l) was intratumorally injected, respectively, in the three groups every other day for 2 times. The volume of tumor and the presence of adverse effects were observed. Seven days after treatment, all mice were sacrificed for evaluation of antitumor effect. Tumors, livers and spleens were all harvested for routine pathological examination. At the same time, the way of tumor cell death induced by vp3 in vivo was identified by TUNEL. Results The titers of both adenovirus AdAFPvp3 and adenovirus AdCMV-eGFP were 5?109pfu/ml. The carcinogenesis rate of hepatocellular carcinoma was 100%. Compared with adenovirus AdCMV-eGFP control group and PBS control group, the volume of tumor was diminished evidently in AdAFPvp3 group after intratumoral injection (P