OBJECTIVETo investigate the molecular mechanism of germ cell apoptosis following testicular torsion in rats.

METHODSHealthy male Sprague-Dawley rats (n = 16) were equally randomized into a control and a torsion group and the models of testicular torsion (720 degrees 2 h) were established. Twenty-four hours later, the apoptosis and count of germ cells were determined by flow cytometry, the expressions of Bax, Fas and Fas ligand (FasL) mRNA semiquantitatively analyzed by RT-PCR and the cytochrome C release detected by Western blot.

RESULTSCompared with the control group, there was an obvious increase in the number of apoptotic germ cells, a marked decrease in that of haploid and tetraploid cells and significantly up-regulated expressions of Bax and Fas/FasL mRNA in the torsion group (P <0.01). The Western blot analysis showed that the cytochrome C release was remarkably increased 24 hours after the detorsion. There were significant differences between the two groups (P <0.01).

CONCLUSIONThere are two major signaling pathways of cell apoptosis following testicular torsion, intercellular and intracellular. Up-regulated expressions of the apoptosis-related molecules Bax and Fas/FasL and increased cytochrome C release may play an important role in germ cell apoptosis following testicular torsion in rats.

Animals ; Apoptosis ; Disease Models, Animal ; Fas Ligand Protein ; metabolism ; Flow Cytometry ; Germ Cells ; cytology ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo investigate the protective effect of pentoxifylline on spermatogenesis following testicular torsion/detorsion in rats.

METHODSHealthy male Sprague-Dawley rats (n = 24) were divided into three groups randomly, each comprising 8 rats. In Group I, rats underwent a sham operation. In Group II and III, animals were submitted to unilateral 720 degrees testicular torsion, then detorsion in two hours. Infusion of isotonic saline and pentoxifylline into tail vein was initiated 15 minutes prior to relief of torsion in Group II and III respectively. Twenty four hours later, testes were examined for evidence of germ cell apoptosis by the flow cytometry and the level of total antioxidant capability (T-AOC) and malondialdehyde (MDA) by spectrophotometry.

RESULTSCompared with that of group II, the number of apoptotic germ cell and the level of MDA decreased remarkably in Group III, but T-AOC increased significantly (P <0.01).

CONCLUSIONPentoxifylline provided significant rescue of testicular function after acute experimental torsion.

Animals ; Apoptosis ; drug effects ; Flow Cytometry ; Germ Cells ; drug effects ; pathology ; Male ; Malondialdehyde ; metabolism ; Pentoxifylline ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; drug therapy ; metabolism ; pathology ; Spermatogenesis ; drug effects ; Vasodilator Agents ; pharmacology

OBJECTIVETo investigate the clinical effect of bridging fixation with locking plate for the Seinsheimer type V subtrochanteric femoral fracture.

METHODSFrom March 2009 to September 2014,18 cases of Seinsheimer type V subtrochanteric femoral fracture were treated by open reduction and bridging fixation with locking plate through proximal and distal approach including 16 males and 2 females with an average age of 41 years old ranging from 22 to 67 years old. Among them, 12 cases caused by traffic accident, 5 cases by falling, 1 case by heavy aboving. All cases were fresh and closed fractures. Time between injury and operation was from 4 to 9 days with an average of 6.2 days. Of them, 11 cases were fixed with reverse LISS and the other 7 cases were fixed with anatomical locking plates of proximal femur.

RESULTSThe mean time of operation was 110 min (ranged from 90 to 155 min). The mean blood loss during operation was 425 ml (ranged from 350 to 650 ml) and 16 cases got blood transfusion which was meanly 300 ml. The mean hospital time was 14 days (ranged from 12 to 18 days). The mean duration of followed up was 11.8 months (ranged from 8 to 22 months). The mean time of bone union was 6.6 months (ranged from 5 to 8 months). There was not any complication such as infection, implant failure, hip varus, external rotation deformity of low limb or fat embolism. The Sanders hip scores were 53.22 ± 6.48, the result was excellent in 12 cases and good in 6 cases at the last follow-up.

CONCLUSIONUnder the principle of biological osteosynthesis, treatment of Seinsheimer type V subtrochanteric femoral fracture with bridging locking plate fixation has such advantages as high mechanism, less interference of blood supply, stable fixation and little complication. It is a safe and idea way for the treatment of the Seinsheimer type V subtrochanteric femoral fracture.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Hip Fractures ; surgery ; Humans ; Male ; Middle Aged

OBJECTIVETo compare the effects of close reduction combined with minimally invasive percutaneous plate osteosynthesis for distal fracture of tibial shaft.

METHODSFrom March 2009 to May 2013, there were 124 patients (89 males and 35 females, 30 of them were injured in a traffic accident and 94 were falling down in daily life, the ages ranging from 21 to 81 years old) who suffered from distal fracture of tibial shaft. Sixty-six patients (48 males and 18 females, 45 cases of type A, 12 cases of type B and 9 cases of type C) were treated with close manipulative reduction combined with minimally invasive percutaneous plate fixation. After close reduction, a minimal incision was made and a anatomic plate was inserted just along the medial tibia periostea, and then the fracture was fixed without fracture exposure. The other 58 patients (41 males and 17 females, 41 cases of type A, 10 cases of type B and 7 cases of type C) were treated with conventional open reduction and internal fixation. Length of the incision, operating time, early postoperative pain (recorded using the Visual Analog Scale score) and the outcome results (recorded using the evaluation standard of Johner-Wruhs) were compared.

RESULTSThe length of incision was meanly (7.34 ± 1.42) cm in MIPPO group and (21.82 ± 2.35) cm in ORIF group; operation time was (44.48 ± 10.00) min in MIPPO group and (59.42 ± 11.84) min in ORIF group. The postoperative radiographs verified good position of all screws and satisfactory bone fracture reduction in both groups. All the patients were followed up, and the duration ranged from 10 to 24 months (mean 15.2 months) in both groups. In MIPPO group,only one patient had delayed union and got union after Chinese herb therapy. The other 65 patients got bony union during 15 to 20 weeks. While in ORIF group, 3 patients suffered from nonunion and received reoperation with bone grafting, and 4 patients got bone infection.

CONCLUSIONTreatment of distal fracture of tibial shaft, combined with close reduction and MIPPO technique, has the advantages such as less invasion, less damage of blood supply, simplified procedure of operation and higher union rate, which is an ideal methods and is accordant to the biological demand.

Adult ; Aged ; Aged, 80 and over ; Bone Plates ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Tibial Fractures ; therapy

OBJECTIVETo investigate the biomechanical differences between the surgery and adjacent segments of intervertebral discs in the lower lumbar spine, which were implanted with Coflex into the segments of L4, and L5S1, respectively.

METHODSThree finite-element models (the model of the intact lower lumbar sacrum,the L4.5 and L5S1 segments implanted by Coflex) were developed, respectively. According to the spinal three-column loading theory, three models were forced by the physiological loads of upright standing, flexion and extension. The stress of the different areas of the disc annulus, the changes of intervertebral dorsal height and the degree of nucleus pulposus pressure were compared and analyzed.

RESULTSCoflex implanted into the L4.5 and L5S1 segments in compression and extension could both decrease the stress of the posterior area of intervertebral disc in the surgery segment, resist the changes of the intervertebral disc dorsal height and reduce the perssure of nucleus pulposus. Furthermore, the stress of the L5S1 segment decreased when Coflex fixed the L4.5 segment in extension. However, when Coflex fixed the L5S1 segment, the stress of L4.5 segment had no significant changes.

CONCLUSIONCoflex fixing the L4,5 and L5S1 segments can effectively decrease the stress of the surgery segmental discs, respectively. Furthermore, Coflex fixing L4,5 segment may play a biomechanical role in reducing the stress of L5S1 segment.

Adult ; Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Internal Fixators ; Intervertebral Disc ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Stress, Mechanical

OBJECTIVETo explore the inhibitory effect of combination of lamivudine with thymosin alpha1 (Talpha1) on the replication of duck hepatitis B virus (DHBV).

METHODSPeking ducks of 1 d old were challenged with DHBV-positive serum and used as a duck hepatitis B model. After treated with lamivudine for three months, the ducks were randomly grouped and treated with or without Talpha1 for 8 d. Serum DHBV titrate was observed by semi-quantitative PCR, and inflammation and degeneration of hepatocytes were observed by pathology examination.

RESULTSThe serum DHBV titrate was significantly reduced (4483.2+/-5193.4 compared with 9351.8+/-5059.6) after lamivudine treatment, and it was reduced more significantly(1692.2+/-589.2) after combination treatment with Talpha1. Lamivudine reduced the degeneration degree of hepatocytes (3.2+/-0.8 compared with 4.6+/-0.5) and the inflammation degree of liver (6.2+/-3.3 compared with 8.6+/-2.8). The combination treatment with Talpha1 increased liver inflammation degree (9.0+/-5.2).

CONCLUSIONBoth Talpha1 and lamivudine may reduce the replication of DHBV in Peking ducks and combination treatment may have the better anti-virus effect and enhance immune response in liver.

Animals ; Animals, Newborn ; Antiviral Agents ; therapeutic use ; Cells, Cultured ; Drug Therapy, Combination ; Ducks ; Hepadnaviridae Infections ; drug therapy ; Hepatitis B Virus, Duck ; drug effects ; genetics ; Hepatitis, Viral, Animal ; drug therapy ; Hepatocytes ; virology ; Lamivudine ; therapeutic use ; Thymosin ; analogs & derivatives ; therapeutic use ; Virus Replication ; drug effects

Accidents, Occupational ; statistics & numerical data ; China ; epidemiology ; Humans

OBJECTIVETo express the domain III of DENV II envelop protein in Escherichia coli, obtain the purified recombinant protein and identify its immunoreactivity.

METHODSSuckling mice were inoculated with live DENV II in the brain. The total RNA was extracted from the brain of the infected mice, and the envelope protein DNA fragment was amplified by RT-PCR and ligated into pMD 18-T to construct pMD 18-T-DV2-E. The domain III DNA fragment of the envelope protein was amplified by PCR with pMD 18-T-DV2-E as the template and cloned into pET-32a(+) to construct the expression plasmid pET-32a(+)-DV2-E-DIII. The recombinant plasmid was transformed into E.coli BL21(DE3) and induced by IPTG, and the expressed products were analyzed by SDS-PAGE and Western blotting.

RESULTSAfter RT-PCR amplification, a specific DNA fragment of about 1.5 kb was obtained and ligated into pMD 18-T to construct pMD 18-T-DV2-E. With pMD 18-T-DV2-E as the template, the domain III DNA fragment about 320 bp in length was amplified and the expression plasmid pET-32a(+)-DV2-E-DIII was successfully constructed. After induction with IPTG, a specific soluble protein with a relative molecular mass of 29000 was obtained and the expression product accounted for 52.50 percent; of the total protein of the cell lysate. Western blotting demonstrated reactivity of the recombinant protein with His-Tag McAb and DENV (Type I-IV) McAb.

CONCLUSIONThe recombinant plasmid can be highly expressed in E.coli BL21(DE3) in a soluble form and the recombinant protein can react with DENV (Type I-IV) McAb.

Animals ; Dengue Virus ; genetics ; isolation & purification ; Escherichia coli ; metabolism ; Genetic Vectors ; Mice ; Mice, Inbred Strains ; Plasmids ; Protein Interaction Domains and Motifs ; Viral Envelope Proteins ; genetics ; isolation & purification

OBJECTIVESTo investigate the relationship between germ cell apoptosis and expression of Bcl-2 and Bax in experimental torsed/detorsed testes of the adult male rats.

METHODSThirty healthy male Sprague-Dawley rats were divided into torsion group (n = 15) and control group (n = 15) randomly. Animals were submitted to unilateral 720 testicular torsion, then detorsion were done in two hours. Three days later, the evidence of germ cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique. The expression of Bcl-2 and Bax were detected by immunohistochemical method.

RESULTSIn the torsed testes, the apoptosis index of germ cell and Bax expression significantly increased compared with that in the control group (P < 0.01) while Bcl-2 expression obviously decreased (P < 0.01). The apoptotic cells were mostly pechytene spermatocytes and round spermatides.

CONCLUSIONSThe germ cell apoptosis is highly associated with expression of Bcl-2 and Bax in experimental testicular torsion. Bcl-2/Bax plays an important role in germ cell apoptosis.

Animals ; Apoptosis ; Gene Expression ; Germ Cells ; pathology ; Male ; Proto-Oncogene Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; pathology ; bcl-2-Associated X Protein

OBJECTIVETo investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) on the differentiation and mineralization of osteoblast (OB). METHODS (OB) was isolated from the skull of 10 newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified, different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction ([Chinese characters: see text]) of different phase (rats were fed with medicine three days or five days after last fed with medicine one hour or three hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected.

RESULTSThe secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum, the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction ([Chinese characters: see text]) three days or five days, after one hour of last fed with medicine.

CONCLUSIONThe pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) three days, after one hour of last fed, with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Diabetes Complications ; drug therapy ; Diabetes Mellitus ; drug therapy ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Female ; Humans ; Male ; Osteoblasts ; drug effects ; enzymology ; physiology ; Osteoporosis ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Serum ; drug effects ; metabolism