BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation.
OBJECTIVE:To construct the targeting short hairpin RNA plasmid vector expressing TERT gene from astrocytes by using pLentilox3.7.U6.
METHODS:By using two sequences from TERT gene, we synthetized sense and antisense strand template sequences of RNA interference molecular in vitro, and then obtained the complementary strands through annealing procedure. We connected the strands with pLentilox3.7.U6 that was sequenced and transfected into the Escherichia coli. In the end, we tested its effect of reducing the TERT gene expressing by using cultured astrocytes from rat spinal cord in vitro through western blot and immunofluorescence technique.
RESULTS AND CONCLUSION:Western blot and immunofluorescence assay showed that, compared with the control group, the interference groups had a lower TERT expression in astrocytes. The targeting short hairpin RNA plasmid vector expressing TERT gene is useful to reduce the TERT gene expression. The targeting short hairpin RNA plasmid vector expressing TERT gene is valid for us to do the further test learning the mechanism of astrocytes in spinal cord injury.

Aged ; Angina Pectoris, Variant ; physiopathology ; Coronary Vasospasm ; physiopathology ; Coronary Vessels ; physiopathology ; Humans ; Male

Emergencies ; Environmental Exposure ; standards ; Humans ; Maximum Allowable Concentration

BACKGROUND:Telomerase reverse transcriptase plays an important role in telomerase activation, and lentiviral vectors carrying human telomerase reverse transcriptase that can inhibit astrocyte expression are rarely reported to have effects on spinal cord injury. OBJECTIVE:To transfect rat astrocytes with telomerase reverse transcriptase gene lentiviral vectors and to observe the effect of telomerase reverse transcriptase gene lentiviral vectors on apoptosis of astrocytes. METHODS: Astrocytes from rats were subject to primary culture and subculture, and then transfected with siRNA lentiviral vector carrying telomerase reverse transcriptase gene (siRNA transfection group), simple lentiviral vector (lentiviral vector group) and nothing (blank group), respectively. Then, the transfection efficiency and apoptosis in different time periods after transfection were determined. RESULTS AND CONCLUSION:The transfection efficiency was up to 85%-90% after siRNA lentiviral vector and simple lentiviral vector transfection. The apoptosis rate of astrocytes in the siRNA transfection group was 50%-60% at 24-48 hours after transfection, but there was no significant change in the other two groups. These findings suggest that siRNA lentiviral vectors carrying telomerase reverse transcriptase gene can accelerate astrocytes apoptosis.

A patient was treated by multiple dental implants,the implanted 6 was adjucent to impacted 8 .Immediately after implanta-tion,4 month and 3 year after implantation the distace between 8 and 6 implant central line was 4.4,3.2 and 2.5 mm,the angle between 8 long axis and 6 implant central line was 42.3°,45.5°and 50.3°.Then 8 was extracted.

OBJECTIVETo investigate the factors influencing the pregnancy outcomes of artificial insemination with donor sperm (AID), improve the pregnancy rate, and evaluate the safety of the offspring.

METHODSWe retrospectively analyzed 7,761 cycles of AID for 5,109 infertile couples performed between July 1, 2005 and June 30, 2013 in the Center of Reproductive Medicine of Shenyang No 204 Hospital, the outcomes of pregnancy, and the incidence of birth defects.

RESULTSTotally, 2 252 clinical pregnancies were achieved by AID, in which the pregnancy rate per cycle was 29. 02% and the cumulative pregnancy rate was 44. 08%. The clinical pregnancy rate was remarkably higher in the females of ≤ 35 years than in those of > 35 years old (30.31% vs 20.18%, P < 0.01), in the women with < 5-year infertility than in those with > 5-year infertility (30.83% vs 28.16%, P < 0.01), and in the patients of the ovarian stimulation group than in those of the natural cycle group (33.22% vs 28.68%, P < 0.01) The clinical pregnancy rate was the highest in the first treatment cycle (29.87%), with statistically significant difference from the fourth cycle (23.61%) (P < 0.05), but not between the other cycles (P > 0.05). There were 28 cases of birth defects in the offspring (1.40%), including 6 cases (21.43%) involving the cardiovascular system, 4 (14.29%) involving the musculoskeletal system, 3 (10.71%) involving the urogenital system, 3 (10.71%) involving the central nervous system, 2 cases (7.14%) of cleft lip and palate, 2 (7.14%) involving the respiratory system, 2 (7.14%) involving the gastrointestinal digestive system, and other anomalies.

CONCLUSIONFemale age, infertility duration, and ovarian stimulation treatment are important factors influencing the clinical pregnancy rate of AID. Artificial insemination with cryopreserved donor sperm does not increase the incidence of birth defects, which is considered as a relatively safe technique of assisted reproduction.

Adult ; Cryopreservation ; Female ; Humans ; Infertility ; Insemination, Artificial, Heterologous ; methods ; Male ; Maternal Age ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Semen Preservation ; methods ; Spermatozoa ; Time Factors

OBJECTIVETo compare therapeutic effects between arthroscopic medial retinaculum plication and plaster external fixation for the treatment of acute patellar dislocation.

METHODSFrom February 2006 to October 2012,29 patients with acute patellar dislocation were divided into two groups: operation group and non-operation group. The patellar dislocation duration was 2 weeks. In operation group, there were 7 males and 10 females, with an average age of (16.2 ± 6.2) years old, and the patients were treated with arthroscopic medial retinaculum plication. In non-operation group, there were 5 males and 7 females,with an average age of (16.3 ± 5.0) years old,and the patients were treated with plaster external fixation. The Kujala scores, patellar tilt angle measured on CT film, apprehension test and recurrence rate of patellar instability were observed before and 1 year after treatment.

RESULTSIn operation group, the pre-treatment and post-treatment patellar tilt angles had no statistical difference, but the post-treatment Kujala score was lower than that of pre-treatment; while in non-operation group, the post-treatment patellar tilt angle was larger than that of pre-treatment, and the post-treatment Kujala score was lower than that of pre-treatment. At 1 year after treatment, the patellar tilt angle (21.2 ± 5.3) of patients in non-operation group was larger than (13.5 ± 3.5) of operation group, and the Kujala score 73.3 ± 10.5 of patient in non-operation group was lower than 84.1 ± 5.6 of operation group.

CONCLUSIONDuring 1 year after operation, arthroscopic medical retinaculum plication is a more effective treatment for acute patellar dislocation compared with plaster external fixation.

Adolescent ; Adult ; Arthroscopy ; Case-Control Studies ; Casts, Surgical ; Child ; Female ; Fracture Fixation ; Humans ; Male ; Patellar Dislocation ; surgery ; therapy ; Patellar Ligament ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the correct method of bone resection and posterior capsular soft tissue releasing in total knee arthroplasty (TKA) for the patients with rheumatoid arthritis with stiff knee in flexion.

METHODSFrom November 2009 to January 2012,15 patients with rheumatoid arthritis with stiff knee in flexion underwent primary TKA and releasing of the posterior soft tissues. There were 7 males and 8 females,aged 22 to 75 years old (58.7 years old on average). The preoperative range of movement(ROM) was (3.2 ± 1.7)°. According to Knee Society score (KSS) criterion, the preoperative clinical score was 23.3 ± 12.5 and functional score was 35.2 ± 9.8. Based on the correct osteotomy, effective releasing of posterior structures was used for different degrees of flexion contracture during the TKA procedure.

RESULTSAll the patients were followed up, and the average duration was 2.3 years (1.6 to 3 years). At the latest follow-up,the KSS clinical score was 81.7 ± 6.5 and functional score was 82.8 ± 9.3. The flexion and extension ROM of the knee joint was (103.5 ± 13.1). Three knees remained 50 flexion contracture deformity, but the function of the affect knees was good.

CONCLUSIONThe effective releasing of the soft tissue of posterior capsule is a major management for correction of the flexion contracture in TKA. The correct releasing of posterior structure can not only achieve fundamental gap of TKA but also effectively avoid bone over-resection.

Adult ; Aged ; Arthritis, Rheumatoid ; complications ; physiopathology ; surgery ; Arthrogryposis ; surgery ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Joint Capsule Release ; methods ; Male ; Middle Aged ; Range of Motion, Articular

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection

BACKGROUND:A large number of studies have confirmed that anterior approach and posterior approach for multilevel cervical spondylotic myelopathy were effective, but there is stil no conclusion in which one is better.
OBJECTIVE:To systematical y assess the clinical effectiveness and safety of anterior approach versus posterior approach for multilevel cervical spondylotic myelopathy.
METHODS:The databases such as The Cochrane Library (Issue 3, 2013), PubMed (from 1966 to March 2013), OVID (from 1950 to March 2013), EMbase (from 1966 to March 2013), Chinese Biomedical Literature Database (from 1978 to March 2013), WanFang Database (from 1998 to March 2013), China National Knowledge Infrastructure (from 1999 to March 2013) were electronical y searched and five relevant journals were searched by hand to col ect the randomized control ed trials or non-randomized control ed trials about the clinical effectiveness and safety of anterior approach versus posterior approach for multilevel cervical spondylotic myelopathy. Two reviewers independently screened the literature according to the inclusive and exclusive criteria, extracted the data, and assessed the methodological quality of included studies. Then the meta-analysis was performed by using RevMan5.2 software.
RESULTS AND CONCLUSION:A total of 11 control ed trials involving 814 patients were included. Meta-analysis results showed that, compared with posterior approach, postoperative Japanese Orthopaedic Association scores were better (P<0.000 01), improvement rate of neurological function was higher (P=0.000 3), the incidence of C5 root palsy was lower (P=0.007), but operation time was longer (P<0.000 01), amount of intraoperative bleedin g was larger (P=0.000 7), incidence of adjacent segments degeneration was higher (P=0.01), incidence of postoperative complications was higher (P<0.000 01) and the rate of secondary surgical procedures was higher (P=0.003) after anterior approach. Additional y, there were no differences between the two groups in the cervical range of motion (P=0.56). For quantity limitation and low methodological quality of included studies, this conclusion stil needs to be further proved by performing more high-quality and large-scale randomized control ed trials.