To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

OBJECTIVETo observe the application effect of minimally invasive decompression, bone graft implantation and metal trabecular bone reconstruction system for early stage osteonecrosis of femoral head and discuss the treatment of hip-salvage operation in early stage osteonecrosis of femoral head;

METHODSFrom January 2010 to June 2011, 50 patients (62 hips) Which were osteonecrosis of femoral head of early stake,were treated with minimally invasive decompression, bone graft implantation and metal trabecular bone reconstruction system, including 31 males (40 hips), 19 females (22 hip) with an average age of 36.2 years old ranging from 22 to 54 years old. The course of disease was from 6 to 15 months (averaged 10.5 months). Among them, 19 cases (23 hips) were steroid-induced, 25 cases (33 hips) were alcohol-induced, 6 cases (6 hips) were idiopathic; According to ARCO stage, 28 hips were at stage I, 34 hips were at stage II. All of them were diagnosed as femoral head necrosis by imaging examination before operation. Then each patient was followed to assess by Harris hip score, curative effect, and conduct the femoral head survival analysis during the postoperation.

RESULTSAll patients had finished operation, the operation time was between 30 and 85 min, intraoperative blood loss was 50 to 220 ml, and 47 cases (58 hips) were follow-up from 24 to 46 months with an average of 34.05 months. As compared with preoperative, the Harris hip score at the last follow-up was improved, the difference was statistically significant (P<0.01). The Harris hip score, curative effect and survival time of femoral head in ARCO stage I was superior to these in ARCO Stage II, the difference was statistically significant (P< 0.05).

CONCLUSIONEffect of minimally invasive decompression,bone graft implantation combine with the metal trabecular bone reconstruction system for early stage osteonecrosis of femoral head was good,it could significantly improve the Harris hip score, increase the femoral head survival time, delay the hip replacement, and performance better in ARCO stage I.

Adult ; Bone Transplantation ; Decompression, Surgical ; Female ; Femur Head ; injuries ; pathology ; surgery ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prostheses and Implants ; Reconstructive Surgical Procedures ; Young Adult

OBJECTIVETo observe the curative effect of porous tantalum rod and Gugutou Huaisiyu Capsule (GHC) for steroid-induced osteonecrosis of femoral head (SONFH).

METHODSA total 60 hips of 50 SONFH patients were randomly assigned to the treatment group and the control group according to grouping time, 25 in each group (30 hips). Patients in the control group were implanted with porous tantalum rod, while those in the treatment group additionally took GHC (5 pills each time, three time per day for 2 successive months; and then twice per day for 4 successive months). Then all patients were followed-up to observe Harris hip score. The curative effect and the femoral head survival time were assessed.

RESULTSA total of 49 patients (59 hips) were followed-up. The Harris hip score of the two groups at the final follow-up was significantly improved after treatment, with statistical difference when compared with before treatment (P < 0.01). Besides, it was higher in the treatment group than in the control group. The curative effect and the survival time were superior in the treatment group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSPorous tantalum rod combined GHC got better effect in treating SONFH. It could significantly improve the function of affected hips and prolong the survival time of femoral head.

Capsules ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Femur Head Necrosis ; drug therapy ; Humans ; Prostheses and Implants ; Steroids ; adverse effects ; Tantalum

Objective To analyze the surgical technique of extraperitoneal laparoseopic radical prostatectomy with neurovascular bundle preservation and its effect on micturition control and sexual function in patients with prostate cancer. Methods Twenty-two patients with a median age of 66. 9 years underwent extraperitoneal laparoscopic radical prostatectomy with neurovascular bundle preservation from October 2005 to March 2009 in our hospital and Shanghai Changzheng hospital. All patients had no erectile dysfunction. According to the pathology, there were 2 cases of PIN high grade, 4 cases of Tla-b, 9 cases of Tlc, 4 cases of T2a-b, 2 cases of T2e and 1 case of T3a. Results All operation were successful and no case was conversed to open surgery. The average operation time was 225 min and the mean blood loss was 850ml. The amount of blood loss in 1 case was 1200 ml because of injuring deep dorsal vein of penis. 4 cases received blood transfusion, gatheters of all patients were removed 2 weeks after operation, and three cases with mild incontinence regained continence of urine after 6 months follow-up. No patient occurred postoperative vesicourethral anastomotic leakage of urine. The surgical margin of 1 case was positive. No biochemical index of prostatl cancer recurred during postoperative follow-up of 10-24 months (with an average of 16 months). 20 cases had sexual function recovery after operation. Conclusions The extraperitoneal laparoscopie radical prostatectomy with neurovascular bundle preservation for prostate cancer is a safe and effective treatment method with less bleeding, small damage and less complications. It can guarantee the maximum of micturition control and sexual function, which is worthy to be popularized in the clinic.

AIMTo explore the effects of heme- heme oxygenase-1 (HO-1)-carbon monoxide(CO)-cyclic GMP (cGMP)on aortic vascular reactivity in endotoxemic rats and its molecular mechanism.

METHODSBy using isolated vascular ring tension detecting technique, cumulative responses of thoracic aortic rings (TARs)to phenylephrine (PE) were measured at 6 h after lipopolysaccharide administration. Effects on contractile responses to PE were measured under which the TARs were incubated with hemin (He, donor of CO), zinc-protoporphyrin-IX(ZnPP-IX, selective inhibitor of HO-1), or methylene blue (MB, inhibitor of guanylyl cyclase), respectively. The content of CO and the activity of HO-1 were measured. The protein and the gene expression of HO-1 were examined by Western blot and RT-PCR.

RESULTSContractile responses of TARs to cumulative doses of PE were depressed by pretreated with LPS. The hyporesponsiveness was partly reversed by incubation with ZnPP-IX and was restored to normal by incubation with MB in endotoxemic rats. Incubation with He could contribute to the vascular hyporeactivity. The content of CO, the activity and the protein and the gene expression of HO-1 were significantly increased in aorta of endotoxemic rats.

CONCLUSIONLPS could induce the HO-1 mRNA and the protein expression, the activity of HO-1 increase in aorta, lead to active the pathway of heme-HO-1-CO-cGMP, which is one of the important mechanisms of the vascular hyporeactivity in endotoxemic rats.

Animals ; Aorta ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Cyclic GMP ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Phenylephrine ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVEThe purpose was to analyze the effects of three sterilization methods (dry heat sterilization, steam sterilization, and chemical sterilization) on the corrosion of dental fissure bur.

METHODS200 dental fissure burs were distributed to 10 groups. Bending strength, elastic modulus, and torsional strength were measured by bending and torsional instrument and calculated with special designed software. Among the three sterilization methods, the steam sterilization group showed the most evident.

RESULTSThe corrosion was most severe in steam sterilization group, followed by chemical sterilization, dry heat sterilization. With the sterilization time increased, bending strength, elastic modulus, and torsional strength decreased respectively. Of the three sterilization methods, the mechanical properties were decreased most evidently by steam sterilization, followed by chemical sterilization and dry heat sterilization.

CONCLUSIONIt is proved that the bending strength, elastic modulus and torsional strength have a tight relationship with the corrosion of dental fissure burs. The corrosion was most severe in steam sterilization group, followed by chemical sterilization, dry heat sterilization. In regards of the corrosive effect, the dry heat sterilization might be the best way to sterilize the dental fissure burs.

Dental Fissures ; Dental High-Speed Equipment ; Dental Instruments ; Steam ; Sterilization

Adolescent ; Adult ; Antigens, CD34 ; metabolism ; Antineoplastic Agents ; therapeutic use ; Benzamides ; Chondroma ; drug therapy ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Gastrectomy ; Gastrointestinal Stromal Tumors ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Imatinib Mesylate ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Neoplasm Recurrence, Local ; Neoplasms, Multiple Primary ; drug therapy ; metabolism ; pathology ; surgery ; Piperazines ; therapeutic use ; Pneumonectomy ; Proto-Oncogene Proteins c-kit ; metabolism ; Pyrimidines ; therapeutic use

The aim of the present study was to explore the effect of cholecystokinin octapeptide (CCK-8) on [Ca(2+)](i) and its signal transduction mechanism in isolated guinea pig cardiomyocytes. [Ca(2+)](i) was measured by laser scanning confocal microscopy in single ventricular myocytes which were dissociated by enzymatic dissociation method and loaded with Fluo 3-AM. The changes in [Ca(2+)](i) were represented by fluorescent intensity (F(i)) or relative fluorescent intensity (F(i)/F(O)%). The results obtained are as follows. (1) In the normal Tyrode's solution containing 1.0 mmol/ L Ca(2+), CCK-8 (1-10(4) pmol/L) elicited a rapid and marked increase in [Ca(2+)](i). (2) When cardiomyocytes were pretreated with the Ca(2+) chelator EGTA (3 mmol/L) and Ca(2+) channel antagonist nisoldipine (0.5 micromol/L) for 5 min, CCK-8 (10(2)pmol/L) caused a slow and small increase in [Ca(2+)](i) (p< 0.01). (3) Pretreatment with the nonselected CCK- receptor (CCK-R) antagonist proglumide (6 micromol/L) or the tyrosine kinase inhibitor genistein (1 micromol/L) for 5 min could inhibit the increase of [Ca(2+)](i) induced by CCK-8 (10(2) pmol/L) (p<0.01). The results suggest that CCK-8 increases the [Ca(2+)](i) via activating the receptor-operated Ca(2+) channel and eliciting the influx of Ca(2+) in isolated guinea pig cardiomyocytes, in which tyrosine kinase may be involved.
Animals ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels ; drug effects ; Cell Separation ; Guinea Pigs ; Myocytes, Cardiac ; metabolism ; ultrastructure ; Nisoldipine ; pharmacology ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Sincalide ; pharmacology

OBJECTIVETo explore the causes and mechanisms of children elbow joint injury led to elbow varus.

METHODSOne hundred children with elbow varus causded by elbow joint injuries were retrospectively studied during January 2005 to April 2008 in department of child orthopedics involving 54 males and 46 females with an average age of 9.7 years old ranging from 7 to 16 years. The anterior-posterior and lateral position X-ray films of elbow joint continuous 12 to 18 months (means 15 months) of all the cases were collected. The postoperative X-ray films were observed to analyze the cause of elbow varus.

RESULTSThere were 81 cases caused by epiphyseal dysplasia, growth imbalance, included 47 cases of humeral supracondylar fractures, 16 cases of entire epiphysis separation of distal humerus, 11 cases of humeral lateral condyle fractures, 7 cases of humeral medial condyle fractures. There were 19 cases caused by ulna drift and rotation of the distal fracture end,the ulnaris cortibal bone crushing and collapse involving 11 of humeral supracondylar fractures, 2 of entire epiphysis separation of distal humerus, 5 of humeral lateral condyle fractures, 1 of humeral medial condyle fractures.

CONCLUSIONThere are two main factors of formation for the children elbow varus deformity: the main causes is epiphysial dysplasia after fracture and imbalance growth of epiphysis; the secondary reasons is ulnar inclination of the distal fracture, rotation of the distal fracture, fracture shift such as ulnar bone cortical extrusion collapse.

Adolescent ; Child ; Elbow Joint ; diagnostic imaging ; injuries ; pathology ; surgery ; Female ; Follow-Up Studies ; Fractures, Bone ; diagnostic imaging ; etiology ; surgery ; Humans ; Male ; Retrospective Studies ; Tomography, X-Ray Computed

Objective To analyze the thickness of cornea and corneal epithelium in healthy subjects by optical coherence tomography angiography (OCTA).Methods Totally 100 healthy subjects aged between 20 and 30 years were analyzed by OCTA technique.Using AngioVue OCTA system of retinal imaging mode,and using SSADA algorithm for imaging,the cornea and the corneal epithelium in the central corneal diameter range of 9 mm were measured.The differences of corneal and corneal epithelial thickness in different gender regions were compared.Results In the male and female group,the corneal central total thickness were (559.92 ±33.26) μm and(540.06 ±31.63)μm,and the corneal epithelial thickness were(57.78 ±4.88) μm and(56.88 ±4.57) μm,The total central corneal thickness and central corneal epithelial thickness of the male were greater than those of the female,the difference was statistically significant (t =3.06,2.10;all P ＜ 0.05).The cornea of male was the thickest at S5,S7 and SN9,there were significant differences at S5 and S7 compared with female (t =2.93,2.83;all P ＜ 0.05);The female cornea was the thickest at S5,SN7 and SN9,and the difference was significant at S5 compared with male.The cornea of male subjects was the thinnest at IT,which was statistically significant only at IT5 compared with female subjects in the same area (t =2.02,P ＜ 0.05);The cornea of female subjects was the thinnest at T5,IT7 and T9,which was statistically significant only at T5 and T9 compared with male subjects in the same region (t =2.63,2.20;all P ＜ 0.05);There was significant difference in corneal thickness between male and female at ST (t =3.1 1,2.79,2.33;all P ＜ 0.05).The corneal epithelium was the thickest at IT5,I7,and I9,and the lowest at S5,S7 and S9,and there was no significant difference compared with female in the same region (all P ＞ 0.05).The corneal epithelium of female at the IT5,T7,N9 were the thickest,SN5,S7,S9 were the thinnest;Except for M2 and SN5,there was no significant differences in corneal epithelium between male and female groups (all P ＞ 0.05).Corneal central epithelium accounted for the largest percentage of total corneal thickness,and gradually decreased from inside to outside.Conclusion OCTA can be used to measure the thickness of corneal and corneal epithelial regions.