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.

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.

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

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

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

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.

BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation, however there is rare report addressing the construction of the lentivirus targeted its genes to inhibit its expression in the spinal cord astrocytes.
OBJECTIVE:To construct recombinant lentivirus vector expressing smal interfering RNA against TERT gene and to evaluate its potential for inhibiting the TERT expression.
METHODS:After shRNA-TERT sequence was designed and synthesized, the sequence was amplified by PCR and then connected to plasmid pLentilox3.7U6-hTERT to construct recombinant plasmid. The recombinant plasmid was then transfected to DH5αcel s to screen positive colony, and the sequence was identified. The recombinant plasmid pLentilox3.7U6-TERT was transfected in 293T cel s, generating recombinant lentivirus Le-TERT. The titer of recombinant lentivirus was determined and Le-TERT was transfected into the rat spinal cord astrocytes. The expression of TERT in astrocytes was detected by RT-PCR, western blot and immunofluorescence assay.
RESULTS AND CONCLUSION:The gene sequencing analysis confirmed that, recombinant plasmid pLentilox3.7U6-TERT was successful y constructed. The real-time quantitative PCR, western blot analysis and immunofluorescence assay indicated that, after Le-TERT was transfected in the astrocytes for 4 days, the inhibition rate of TERT mRNA was (63.98±2.6)%, and Le-TERT was lowly expressed in the transfected astrocytes. Recombinant expression vector pLentilox3.7U6-TERT can produce the lentivirus at high titer and effectively inhibit TERT expression in the transfected astrocytes.

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.
Antioxidants ; pharmacology ; Ecosystem ; Free Radical Scavengers ; pharmacology ; Macau ; Plant Extracts ; pharmacology

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