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.

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

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

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

OBJECTIVETo seek a rapid, simple, but effective inquiry method for screening high risk population challenged by schistosomiasis.

METHODSTwo embankment collapsed villages were selected in schistosomiasis epidemic area in Dongting Lake. Information on water exposure was collected through a retrospective study. Data was analyzed by stepwise discriminant analysis.

RESULTSA Fisher's function was established by stepwise discriminant analysis which including 5 variables out of 18. Two hundred and forty-six individuals were discriminated by the function with accuracy, sensitivity and specificity of predicting their current infection status with the results of 87.4%, 84.1% and 89.0% respectively.

CONCLUSIONThe inquiry method might serve as simple, rapid, economic and effective tool for diagnosis in screening high risk population challenged by schistosomiasis in lake communities.

Adult ; China ; epidemiology ; Disasters ; Discriminant Analysis ; Female ; Humans ; Male ; Mass Screening ; methods ; Multivariate Analysis ; Retrospective Studies ; Risk Assessment ; Rural Health ; Sampling Studies ; Schistosomiasis japonica ; epidemiology ; prevention & control ; Sensitivity and Specificity ; Surveys and Questionnaires

OBJECTIVETo study the characteristics of allergic reactions to common aeroallergens in young children with wheezing or allergic diseases by examining the results of skin prick test in children under 5 years old.

METHODSA total of 196 children under 5 years old, from a district of Changsha City sampled between September 1 to December 31, 2010, were assigned into two groups according to the presence of wheezing or allergic diseases: allergen screening (n=102) and control (n=94). Skin prick tests were performed on both groups.

RESULTSThe positive rate of skin prick test in the allergen screening group was 61.8% (63/102), and this was significantly higher than in the control group (9.6%, 9/94; P<0.05). In the allergen screening group, the positive rate of skin prick test in children with both recurrent wheezing and allergic rhinitis was significantly higher than in children with wheezing alone (P<0.05). The frequency of wheezing was positively correlated with a positive skin prick test (r=0.91; P<0.05). The positive rate of skin prick test for mites was significantly higher than for other aeroallergens (24.2% vs 3.5%; P<0.05) in the allergen screening group. Skin prick testing of the children for dermatophagoides farinae showed a higher positive rate than for dermatophagoides pteronyssinus (50.0% vs 14.7%; P<0.05).

CONCLUSIONSWheezing in early childhood may be associated with the occurrence of asthma. Skin prick testing contributes to the diagnosis of allergic diseases and assessment of allergic reactions to aeroallergens in children with wheezing.

Asthma ; etiology ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Sounds ; diagnosis ; immunology ; Rhinitis, Allergic, Perennial ; diagnosis ; immunology ; Rhinitis, Allergic, Seasonal ; diagnosis ; immunology ; Skin Tests

OBJECTIVETo observe the effect of electro-acupuncture (EA) on clinical outcomes and the occurrence of ovarian hyperstimulation syndrome (OHSS) in in vitro fertilization and embryo transplantation.

METHODSTotally 109 patients who routinely received in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) at Reproductive Center were assigned to the control group (56 cases) and the EA group (53 cases) according to even and odd-numbered date. Patients in the control group received controlled ovarian hyperstimulation (COH) referring to GnRH-a long protocol. On the basis of COH, those in the EA group received EA from the day of Gn injection to the day of embryo transfer. Estradiol (E2), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and angiotensin (AT) II were measured in all patients on the day of hCG injection, the day of ovum pick up (OPU), and the day of embryo transfer (ET), respectively. The oocyte retrieval rate, good quality embryo rate, clinical pregnancy rate, the abortion rate, and the occurrence of OHSS were compared between the two groups.

RESULTSCompared with the control group, serum E2 levels on the day of OPU and the day of ET were significantly lower in the EA group (P < 0.05). On the day of OPU levels of VEGF and IL-6 also significantly decreased (P < 0.05). Serum levels of VEGF and IL-6 reached the highest line on the day of hCG in the two groups, and then showed a decreasing trend. Compared with the control group at the same time point, serum levels of VEGF and IL-6 obviously decreased on on the day of OPU, hCG, and ET (P < 0.05). The occurrence of OHSS and the canceling rate of transplant cycle were significantly lower in the EA group than in the control group (P < 0.05).

CONCLUSIONSEA, as an adjunctive therapy, could reduce the occurrence of OHSS in IVF. Besides, it did not decrease good embryo rates and pregnancy rates in IVF-ET, which might be associated with lowering local vascular permeability of ovaries.

Electroacupuncture ; methods ; Embryo Transfer ; Estradiol ; metabolism ; Female ; Fertilization in Vitro ; Gonadotropin-Releasing Hormone ; metabolism ; Humans ; Interleukin-6 ; metabolism ; Ovarian Hyperstimulation Syndrome ; therapy ; Ovulation Induction ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; Vascular Endothelial Growth Factor A ; metabolism

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