Objective:To compare the content of polydatin and emodin between polygonum cuspidatum pieces and dispensing gran-ules to provide theoretical basis for the improvement of preparation process of the granules. Methods:An HPLC method with linear gra-dient elution was used to determine the content of polydatin and emodin in the pieces and dispensing granules, and the difference in the two contents was compared. Results: The content of polydatin and emodin in polygonum cuspidatum pieces met the requirement de-scribed in China pharmacopoeia, which was lower than that in the dispensing granules, and the contents of the two components in the commercial dispensing granules were not equivalent to the labeled amount, which was 1/15 and 1/27 of that in the pieces. Conclu-sion:The transport rate of the traditional preparation process of dispensing granules is with poor efficiency, which should be improved.

AIM: To isolate and culture the adipose-derived stem cells (ADSCs) from guinea pig so as to create bases for the experimental study on treating sensorineural deafness by ADSCs transplantation. METHODS: The experiment was carried out between December 2005 and March 2006 in the Open and Key Laboratory of Clinical Medicine at Henan Universities. The inguinal fat pads were excised from adult male guinea pig weighing 500-750 g. A great quantity of adhesive and natant blood cells were observed under inserted microscope and phosphate buffer was used to wash natant cells repeatedly. Then the cells were cultured in DMEM with 10% fetal bovine serum. Before passage, the cells were washed by a small quantity of phosphate buffer and then cultured in 0.25% trypsin and 0.02% EDTA (2 mL). Then most of the ADSCs presented cytoplasm recovery and round shape, followed by digestion of 2 mL DMEM with feral bovine serum. Cellular suspension was collected to count the cells that were incubated at the density of 2?104/cm2. The morphology of ADSCs was observed constantly. Growth curve of ADSCs at passages 1, 3, 10 were recorded and detected by MTT. Surface markers of ADSCs were detected by flow cytometry. RESULTS: ①The result of primary culture showed that ADSCs at day 2 began to adhesion and more than 90 % of ADSCs at day 7 were approximately fusiform or fibroblast-shaped.②MTT detection showed that ADSCs had the capability to proliferate successively, especially at passage 1, 2, 3. But ADSCs at passage 10 presented decreasing proliferation. After digestion, the cells at passages 1, 3, 10 experienced the latent period (24-48 hours), logarithmic phase (3-5 days) and growth platform phase.③CD105 and CD44 were positive by flow cytometry, whereas CD34 was negative. CONCLUSION: Due to stable growth and rapid proliferation, the ADSCs from guinea pig can be used as the donor cells in the treatment of sensorineural deafness by stem cell transplantation.

Diabetic cardiomyopathy ( DC) is an independent complication of diabetes mellitus accompanied with cardiac metabolic imbalances ( increase of fatty acids and reduction of glucose) ,which would impair cardiac function and structure seriously. Peroxisome proliferator-activated receptors ( PPARs) are ligand-activated transcription factors belonging to the nuclear hormone receptor superfamily,and they could transcriptionally regulate energy metabolism and function. PPARs also play an important role in regulating metabolism in DC heart,and directly or indirectly affect cardiac function and structure.

This study was aimed to compare and investigate protective effects of metformin and rosiglitazone on cardiac dysfunction in experimental diabetic cardiomyopathy (DC) model rats. The experimental DC rat model was induced by feeding high calorie diet plus single intraperitoneal injection of small dosage of streptozotocin (STZ). Then, intragastric administration of metformin (140 mg·kg-1) or rosiglitazone(2 mg·kg-1) was given to DC rats for consecutive 6 weeks. Parameters of general signs, eating amount, blood sugar, blood lipids, heart function, heart structure and lipometabolism of myocardial tissues were measured. The results showed that both metformin and rosiglitazone can obviously improve the myocardial injury of DC model rats and reduce the CK value (P < 0.01). Metformin can obviously increase the cardiac output of DC model rats (P < 0.01). Rosiglitazone can improve the maximum rate of myocardial contraction and diastole of the model rat's left ventricle (P < 0.05). Both metformin and rosiglitazone can decrease interventricular septal thickness (IST) value (P < 0.01). Metformin can obviously im-prove general signs of DC rats, inhibit body weight loss and reduce water intake (P < 0.05). Metformin can obvi-ously reduce the blood sugar level (FBG, GSP, HbA1c and FMN) of DC rats (P < 0.05, or P < 0.01) as well as the concentration of TG (P < 0.01). Rosiglitazone can reduce the concentration of FBG and HbA1c (P < 0.05). Meanwhile, rosiglitazone can significantly reduce the concentration of TG and LDL as well as obviously increase the myocardial FA-β-oxidase (P < 0.05). It was concluded that both metformin and rosiglitazone can recover the cardiac dysfunction and myocardial injury of DC rats on certain level. Metformin showed more effects on eating amount and body weight improvements of DC rats.

The development of body donation remains relatively slow in China.One of its main influencing factors restricted to the development of donation is the anatomy teachers' quality.Lots of volunteers to donate and their families are not satisfied with it,which eventually leads to the failure of the donation.Therefore,enhancing anatomy teachers' quality has become one of the key steps in the donated work.It needs to improve the idea awareness of teachers,to standardize the behavior of teachers,to enhance the role models of teachers,and to carry out a regular humanities education and training to truly realize "people-oriented" goals.

Viable but non-culturable state in bacterial cells as received uncultured microorganism has be- come the fundamental research issues in medical microbiology, epidemiology, general microbial ecology and sanitation quarantine since it was put forward in 1982. When in this state, bacterium are no longer able to grow and form colonies on conventional culture media, but maintain their patheogenicity, may become latent infection escaping detection to threaten enviroment and human security. With the development of modern molecular biology technique and metagenomics, it provide new research method and opportunity for uncul- tured microorganism in the environment. Recently, with the application of the metagenome technique, ge- netic fingerprinting technical etc, it becomes more and more popular. Simultaneously, along with this state become maturity in the laboratory. It provides a new research route for development and exploitation the uncultured microorganism.

Objective To assess the effects of surgical treatment for patients with blepharochalasis and its associated abnormali- ties. Design Retrospective case series, Participants 35 cases (52 eyes) with blepharochalasis in stable phase. Methods The correction of the abnormalities in upper eyelids: after designing the lid crease incision, the redundant eyelid skin and prolapsed fat were excised, and prolapsed lacrimal glands were sutured back into position in 18 cases (36 eyes) and ptosis was treated in 10 cases (16 eyes). The correction of abnormalities in lower eyelids: lower eyelid retraction was corrected in 4 cases (6 eyes). The correction of lateral canthus malformation: lateral canthus rounding was treated in 7 cases (14 eyes), accompanied with or followed by correction of baggy eyelid or ptosis. Main Outcome Measures The shape, location and movement of bilateral eyelids, the location of lacrimal glands and the cir- cumstance of tear secretion. Results During the follow-up of 6～60 months, the appearance and location of bilateral eyelids and can- thuses were satisfying, the function of eyelids were normal, and no dry eye symptoms were found. Two cases (3 eyes) appeared recurrent lacrimal gland prolapse 29 and 36 months postoperation and received surgeries for correction again. There was no recurrence after the second prolapse correction surgery in the follow-up of 18 and 24 months respectively. Conclusions Surgical treatment for patients with blepharochalasis and its associated abnormalities is safe and effective. Its recurrence rate is low.

Objective To explore the clinical curative effect of hollow screw internal fixation for the treatment of distal epiphyseal teenagers tibial fracture.Methods Thirty -nine cases of epiphyseal fractures of the distal tibia were treated in Shanghai Children′s Hospital between February 2005 and September 201 4.This study included 24 boys and 1 5 girls with the average age of 1 3.1 years (ranged 1 0.5 to 1 6.2 years old).Preoperative diagnosis was confirmed by the X -ray films or CT examination.All the cases were closed fractures and manipulative reduction all failed.Surgi-cal reduction was performed when plain radiographs showed the fracture gap of epiphysis board and articular surface were greater than 2 mm or after closed reduction.Under C -arm X -ray machine,ankle axial traction was performed initially.Anatomical reduction underwent according to the bone fracture type.The fractures were fixed with cannulated screws by percutaneous or open approach.Postoperative X -ray or CT confirmed anatomical reduction at articular sur-face and growth plate.All patients had been immobilized with short -leg cast for 4 -6 weeks.The ankle joint function and growth were evaluated by means of American Orthopaedic Foot and Ankle Society(AOFAS)Ankle Hindfoot Scale. Results All patients were followed up for 5 to 91 months with an average duration of 35 months.X -ray films showed that all fractures were bone healing.No uneven articular surface was found and there was no extremity rotation and shorte-ning deformity.Results were evaluated by AOFAS scoring system:excellent in 27 cases,good in 10 cases,and general in 2 cases.All patients were able to participate in the normal physical activities.Conclusions CT with multiplanar recon-struction is a premise to make an accurate diagnosis and to choose a reasonable approach.For failed closed reduction for adolescent distal tibial epiphyseal fractures,the cannulated screws are simple and may have a satisfactory effect.

ObjectiveTo investigate the role of caspase 3 in HMME-induced apoptosis in hypertrophic scar fibroblasts (HSFs).MethodsFibroblasts were obtained from 10 patients with untreated hypertrophic scar,and subjected to a primary culture.After 4 to 6 passages of culture,the HSFs were divided into 3 groups to remain untreated(control group),be treated with HMME followed by photodynamic therapy (HMME-PDT group),or the combination of HMME and Z-DEVD-FMK followed by photodynamic therapy (caspase 3 inhibitor group).At 12 hours after the therapy,HSFs were collected and immunofluorescence microscopy was used to observe the fluorescence intensity of caspase 3 after staining with fluorescein isocyanate (FITC) and popodium iodide (PI),flow cytometry was performed to determine the percentage of caspase 3-positive HSFs and apoptosis rate in HSFs after single staining with FITC and PI respectively.Results The fluorescence intensity of caspase 3 was weak in the control group and caspase 3 inhibitor group,but was strong in the HMME-PDT group.An increased percentage of caspase 3-positive HSFs was noted in the HMMEPDT group compared with the control group and caspase 3 inhibitor group(30.86％ ± 1.21％ vs.3.12％ ±0.28％ and 2.46％ ± 0.18％,t =19.92,21.76,both P ＜ 0.05).The apoptosis rate in HSFs was significantly higher in the HMME-PDT group and caspase 3 inhibitor group than in the control group(30.54％ ± 3.78％ and 10.46％ ± 2.15％ vs.2.45％ ± 0.22％,t =35.90,27.97,both P＜ 0.05),and higher in the HMME-PDT group than in the caspase 3 inhibitor group.ConclusionsThe apoptosis in HSFs induced by HMME-PDT is closely related to the activation of caspase 3,while caspase 3 seems to be dispensable for the apoptosis.

Hela is the cell line of adenocarcinoma of the uterine cervix, and human papillomavirus (HPV) 18 shows positive. We delivered siRNA with target specifically to HPV18 E7 mRNA into nude mice Hela tumor xenografts by nanopatch to inhibit the HPV gene expression, and further to study the superiority, the best action time and concentration of siRNA of using nanopatch to transfer siRNA in vivo. We designed siRNA that target specifically to HPV18 E7 mRNA (siE7) and checked the effect of siE7 in vitro. Tumor xenografts were transfected with siE7 and GenEscort III by nanopatch. Expression of HPV18 E7 mRNA and protein were detected 0 hours, 24 hours, 48 hours, 72 hours after transfection with PT-PCR and Western blot, and the best action time was analyzed using nanopatch to thansfect siRNA in vivo. We transfected GenEscort III and siE7 of Different concentration into tumor xenografts respectively by nanopatch and intraperitoneal injection. Expression of HPV18 E7 mRNA and protein was detected 72 hours after transfection by PT-PCR and Western blot, to analyze the best action concentration of siRNA and the superiority of using nanopatch to thansfect siRNA in vivo. The results proved that SiE7was efficient to inhibit expression of HPV18 E7 mRNA and to advance Hela apoptosis in vitro. SiE7 transfected by nanopatch into xenografts could inhibit effectively expression of HPV18 E7 mRNA and protein. The best action time and concentration of siRNA of using nanopatch to thansfect siRNA in vivo are 72 hour post-transfection and 2 micromol/L siE7. To compare intraperitoneal injection in delivering siRNA in vivo, the effect of nanopatch is very predominant. It can be well concluded that Nanopatch can effectively transfer siRNA in vivo, which can effectively inhibit the HPV gene expression.
Animals ; Apoptosis ; DNA-Binding Proteins ; genetics ; Female ; Gene Expression Regulation, Viral ; HeLa Cells ; Humans ; Mice ; Mice, Nude ; Nanostructures ; administration & dosage ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; RNA, Small Interfering ; administration & dosage ; Transfection ; Uterine Cervical Neoplasms ; therapy ; Xenograft Model Antitumor Assays