Adult ; Arteries ; anatomy & histology ; Asian Continental Ancestry Group ; Female ; Humans ; Larynx ; anatomy & histology ; blood supply ; Male ; Microsurgery

BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfacted into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-βl (TGFβ_1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytas were isolated by use of trypsin digestion method. Vector was PLNCX_2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed_2 double digestion, to build PLNCX_2-RFP. TGFβ_1 gene was amplified from the PGEMT-TGF and connected with PLNCX_2-RFP following double digestion, to build PLNCX_2-TGFβ_1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX_2 group and transfected PLNCX_2-TGFβ_1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supematant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ_1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX_2-TGFβ_1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ_1 and RFP, which showed that the eukaryotic expression vector PLNCX_2-TGFβ_1-RFP had been successfully built as expectation. They were then transfected into packaging calls and cultured, the virus titer was defined as 1×10~6 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent calls formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the calls proliferated actively. NO concentration in the transfected PLNCX_2-TGFβ_1-RFP group was higher than that of transfected PLNCX_2 group (P < 0.05), no difference was significant between control group and transfected PLNCX_2 group. The control group and the group transfected PLNCX_2 showed no TGFβ_1 expression, while TGFβ_1 concentration was (28.08±3.73) ng/L in the transfected PLNCX_2-TGFβ_1-RFP group. PLNCX_2 ratroviral vector-mediated human TGFβ_1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage calls proliferate actively.

The clinicopathological data of 173 cases of inflammatory myofibroblastic tumor (IMT)were retrospectively analyzed.Among them 125 were males and 48 females with a mean age of 47.9 y (5-78 y).The lesions of 117 cases were located in lungs,41 cases in eyes,8 in ileocecum,2 in liver,2 in spleen,1 in abdominal wall,1 in maxillary sinus and 1 in face.The average diameter of lesions were 3.5 cm,ranging from 1.0 cm to 7.0 cm.The clinical manifestations were not specific,depending on the locations of tumor.The imaging examinations were helpful for diagnosis.The prcsence of slender-spindled myofibroblasts and proliferation of fibroblast cells,with infiltration of chronic inflammatory cells were the basic histopathological features of the disease.Combined with the histological characteristics and immunohistochemical staining IMT can be differentiated from other spindle cell tumors.Appropriate surgical resection is the main treatment for IMT.

Objective To investigate the protection mechanism of ulinastatin on bacterial endotoxin-induced acute lung injury. Methods Acute lung injury was induced by Escherichia colilipo-polysaccharide(LPS)5 mg·kg-1·d-1,intratracheally. Twenty SD rats were randomly divided into control group(n=10)and ulinastatin group(n=10). Ulinastatid group received ulinastatin 50 kU·kg-1 ,the control groups received the same amount of 0. 9% sodium chloride solution. Then the expression changes of rat AQP-1 and AQP-5,alveolar wall thickness change and the degree of pulmonary edema were detected. Results After the injection of LPS into the rat,the expression of AQP-1 and AQP-5 in control group were continuously decreased,but those in ulinastatin group decreased were not obvious. The lung wet/dry weight ratio in the control group increased significantly,the not obvious changes in the ulinastain group. The thickness of the alveolar in 24,48,72 h of the control group were(3. 84±0. 68),(6. 32±1. 08),(11. 03±2. 47)μm, respectively,and those in the ulinastian groups were(2. 31±0. 44)(,3. 76±0. 82)(,2. 94±0. 67)μm,respectively. Conclusion The AQP-1 and AQP-5 induced the occurrence of pulmonary edema by changing the cell permeability. Ulinastatin can slow down the process so as to reduce the degree of endotoxin-induced lung injury.

Objective To investigate the safety and obtain parameters of microeoagulation of dorsal root entry zone (DREZ) of cervical cord with bipolar foreps on animal model,and provide histological base for clinical application of treatment of brachial plexus avulsiol pain using microcoagulation of dorsal root entry zone.Methods On the base of swine's weight and spinal cord size in similar to human being,it was chosen to be experimental animal.The right DREZs of cervical cord were microcoagulated with bipolar forceps.The swines were fed in normal way.Their activities were observed.The mass change of the cervical cord segment were observed after 3 weeks and the cervical cord segment was fixed with 10% fromalin,paraffin sliced,HE dying.Coagulating space,depth and width were measured under microsope.The coagulating parameter were adjusted according to measuring outcome in order to achieving a most avaliable parameter.Results All post-op swine survived.When the microcogulation were made with bipolar forceps adopted following parame- ters:The distance of between the polar was 2.0 mm;The diameter of polar was 0.3 mm.The inserting depth 2 mm,the coagulated power 18 watt,the coagulated time was 2 second,then the width of lesions of DREZ in cross section was 1.15 mm and the depth of lesions was 3.10 mm,which was consistent with the area of hu- man DREZ of cervical cord.Conclusion The experiment on swine suggested,microcoagulation of DREZ by bipolar forceps is safe and no mortal complications when the testified parameters are adoped,and can achieve the area of DREZ of cervical cord in human.

Objective： The current study was designed to evaluate the treatment results of nasopharyngeal carcinoma by conformal radiotherapy. Methods： From September 1997 to February 2000, a total of 41 cases of recurrent nasopharyngeal carcinoma were treated by conformal radiotherapy. Among them, 27 cases were treated by late-course hypofraction conformal radiotherapy with the dose of 20- 24 Gy at 80％ isodose, 4- 7 Gy per time, following routine radiotherapy of 39 Gy/30f/3weeks. Fourteen cases received total-course conformal radiotherapy with the dose of 59.8 Gy/46f/4.6weeks at 90％ isodose. Results： The reference isodose curves conform to the target in three dimensions in conformal radiotherapy. The acute radiation mucositis, cranial nerve paralysis and trismus incidence were similar in two groups, however, in the hypofractionation group 22％ developed necrosis of nasopharyngeal mucosa, 3.7％ developed reduced optic nerve lesion, whereas none of the hyperfractionation group had these complications. The tumor response rate were both 93％ for two groups. Conclusion： Hyperfraction conformal radiotherapy is effective in the treatment of recurrent nasopharyngeal carcinoma and has relatively lower incidence of radiation complication.

Objective To construct the bioartificial liver by bone mesenchymal stem cell (BMSC) and alginate scaffold. Method Alginate scaffold was used as the cell carrier for the cultivation of BMSC and the differentiation from BMSC into hepatic like cells was induced by the cell factors of HGF, EGF and FGF-4 in the scaffold in vitro. The compatibility of the cells and the scaffold was observed by microscopy and the function of the differentiatd cells was tested. The gene of AFP and ALB was detected by RT-PCR. The secretion of ALB and the urea synthesis of the cells were tested by ALB kit and urea kit respectively. The glycogen synthesis and the CK-18 was tested by the glycogen stanning method and the immunofluorescence test. Results BMSC was able to attach, grow and proliferate well in the alginate scaffold, the well compatibility was observed by microscopy. ALB and urea were detected in the cultivating medium, the gene of ALB and AFP was identified by RT-PCR. The glycogen synthesis ability and the expression of CK-18 were induced during the differentiation. Conclusion The three dimensional atginate scaffold exhibited well compatibility with BMSC, BMSC could be differentiated into the hepatic like cell in the scaffold. BMSC and the alginate scaffold could be used to construct the bioartificial liver for the hepatic tissue engineering.

Differences in gene expressions were compared by cDNA microarray in skeletal muscle of type 2 diabetic rats and normal rats.In diabetic rats,157 genes were down-regulated and 100 genes up-regulated. Some of these genes were related to insulin resistance,glucose and lipid metabolism.

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

Fifty-eight samples belonging to 7 species of Arisaematis Rhizoma and its adulterants were collected. The ITS2 locus was employed as a DNA barcode and amplified, sequenced and assembled for all of the collected samples. Then, ITS2 sequences have been annotated using HMM-based method. The intra- and inter-specific variations were calculated and NJ tree was constructed using MEGA 6.0 software. The results showed that inter-specific K2P distances were significantly larger than intra-specific distances for all of the three origin species of Arisaematis Rhizoma. Furthermore, three origin species, Arisaema amurense, A. erubescens and A. heterophyllum, can be respectively formed to be a single branch with high bootstrap values. It is concluded that ITS2 can be used to correctly identify Arisaematis Rhizoma from its adulterants and the application of ITS2 in the identification of traditional Chinese medicine has an important prospective.
Arisaema ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics