SHIV-CN97001 played an important role in assessing the immune effect and strategy of the AIDS vaccine which included genes of the predominant prevalent HIV-1 strain in China. In this study, SHIV-CN97001 was in vivo passaged serially to construct pathogenic SHIV-CN97001/rhesus macaques model. To identify variation in the gp120 region of SHIV-CN97001 during passage, the fragments of gp120 gene were amplified by RT-PCR from the plasma of SHIV-CN97001 infected animals at the peak viral load time point and the gene distances (divergence, diversity) were calculated using DISTANCE. The analysis revealed that the genetic distances of SHIV-CN97001 in the third passage animals were the highest during in vivo passage. It had a relationship between viral divergence from the founder strain and viral replication ability. The nucleic acid sequence of the V3 region was highly conservative. All of the SHIV-CN97001 strains had V3 loop central motif (GPGQ) and were predicted to be using CCR5 co-receptor on the basis of the critical amino acids within V3 loop. These results show that there was no significant increase in the genetic distance during serial passage, and SHIV-CN97001 gp120 gene evolved toward ancestral states upon transmission to a new host. This could partly explain why there was no pathogenic viral strain obtained during in vivo passage.

Aged ; Aged, 80 and over ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Hip Fractures ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control

Objective: To evaluate the efficacy of Tuina (Chinese therapeutic massage) manipulation plus horse-riding squat exercise in treating knee osteoarthritis (KOA) and optimize the combining protocol. Methods: Based on a 2×2 factorial design, 120 eligible KOA patients were randomized into a manipulation group (group A1B2), a manipulation plus horse-riding squat group (group A1B1), a sitting knee-adjustment group (group A2B2 group), and a sitting knee-adjustment plus horse-riding squat group (group A2B1), with 30 cases in each group. The intervention was conducted three times a week, lasting for four weeks. The Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was taken as the major measure for efficacy evaluation (including three component scores, pain, stiffness, and daily function, and total score). Results: The three component scores (pain, stiffness, and daily function) and the total score of WOMAC showed significant differences after the intervention in the four groups (P<0.05). There were significant inter-group differences in the WOMAC stiffness score amongst the four groups after the intervention (P<0.05). In group A1B1, the step length, stride, walking speed, and knee joint flexion angle changed significantly after treatment (P<0.05). After the intervention, the step length changed significantly in group A1B2 (P<0.05), and the walking speed changed significantly in group A2B1 (P<0.05). There were no significant differences in the step length, stride, walking speed, or knee joint flexion angle among the four groups (P>0.05). The extensor peak torque at 180 °/s changed significantly in group A1B2 after treatment (P<0.05). Neither the intra-group nor the inter-group comparisons of the four groups revealed significant differences in the other isokinetic muscle strength parameters (P>0.05). The main effect of manipulation showed significant in affecting the WOMAC pain and total scores (P<0.05). The main effect of horse-riding squat exercise showed significant in affecting the WOMAC pain and stiffness scores (P<0.05). Conclusion: The four treatment protocols all can improve the symptoms of KOA, for instance, relieving pain and stiffness, and enhancing daily function. Group A2B1 produces the most eminent effect in relieving joint stiffness. The main effects of both manipulation and horse-riding squat exercise are significant in reducing pain. Besides, the main effect of horse-riding squat exercise is significant in relieving joint stiffness.

OBJECTIVETo observe the changes of nuclear factor-kappa B (NF-kappaB) in the course of N-methyl-N-nitrosourea (MNU)-induced apoptosis of rat retinal photoreceptor cells and investigate the mechanism of MNU-induced retinal damage.

METHODSA single intraperitoneal injection of 60 mg/kg MNU was given to 50-day-old female rats, which were sacrificed at different intervals after MNU treatment. The retinal damage was examined with optical microscopy and photoreceptor cell apoptosis detected by TUNEL assay. Western blotting was performed to analyze the changes in NF-kappaB.

RESULTSPyknosis of the photoreceptor cell nuclei and disorientation of the outer segment of the photoreceptor layer was observed 24 h after MNU treatment, and the outer nuclear layer and photoreceptor layer were almost completely lost on day 7. Photoreceptor cell apoptosis peaked at 24 h, and in the apoptotic cascade, NF-kappaB p65 protein was only detected 12 and 24 h after MNU treatment, whereas the amount of I kappa B alpha, in contrast, markedly increased in the cytoplasm as well as in the nuclei.

CONCLUSIONMNU-induced retinal damage might be mediated through the signaling pathway of NF-kappaB/I kappa B alpha.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Female ; I-kappa B Proteins ; metabolism ; In Situ Nick-End Labeling ; Methylnitrosourea ; toxicity ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retinal Diseases ; chemically induced ; metabolism ; pathology

OBJECTIVETo study the correlation between morphological variation and gentiopicroside content in cultivated Gentiana manshurica roots and to investigate the feasibility of breeding new varieties of high effective constituent content.

METHODGentiopicroside was determined in 5 morphological types of cultivated G. manshurica roots by HPLC, which are normal (or wild) type, white-flowered type, thick-rooted type, broad-leaved type I and broad-leaved type II.

RESULTAmong different types gentiopicroside content is the highest in the roots of thick-rooted type, the contents decrease as following order: normal type, broad-leaved type I white-flowered type and broad-leaved type II, and the gentiopicroside contents in the same type root system are a positive correlation with root ages, as 3-years-age roots > 2-years-age roots > 1-year-age varied with roots.

CONCLUSIONThe contents of effective constituent vary with the morphological variation in cultivated G. manshurica. It is feasible to breed a new variety with high effective constituent with the morphological character as a selecting index.

Genetic Variation ; Gentiana ; anatomy & histology ; chemistry ; classification ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; classification ; Pyrans ; analysis ; Species Specificity

BACKGROUNDPrevious studies have showed that photooxidative stress can lead to down-modulation of nuclear factor-kappa B (NF-kappaB) activity causing apoptosis of cultured photoreceptor cells. This study aimed at investigating whether NF-kappaB was involved in photoreceptor cells apoptosis induced by N-methyl-N-nitrosourea (MNU) in rats.

METHODSA single intraperitoneal injection of 60 mg/kg MNU was given to 50-day-old female rats. At different intervals after MNU treatment, the animals were sacrificed. Retinal damage was examined by a light microscope. The apoptotic index of the photoreceptor cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL). NF-kappaB was analysed by Western blot and Transcriptin Factor Assay Kits.

RESULTSThe pyknosis of the photoreceptor nuclei and the disorientation of the outer segment of the photoreceptor layer was seen after MNU treatment for 24 hours. The outer nuclear layer and photoreceptor layer were almost completely lost at 7 days. Photoreceptor cells apoptosis reached the peaked value at 24 hours. In apoptotic cascade, the protein levels of NF-kappaB p65 were only detected after MNU treatment for 12 and 24 hours in the nucleus. Conversely, the amounts of IkappaBalpha were markedly increased in the cytoplasm as well as in the nucleus. The activity of NF-kappaB p65 in the nucleus was down-modulated in the end.

CONCLUSIONSMNU-induced photoreceptor cell destruction was attributed to the apoptotic process by down-regulating the activation of NF-kappaB p65.

Animals ; Apoptosis ; drug effects ; Cell Nucleus ; metabolism ; Female ; I-kappa B Proteins ; analysis ; physiology ; Methylnitrosourea ; toxicity ; NF-KappaB Inhibitor alpha ; NF-kappa B ; analysis ; physiology ; Photoreceptor Cells ; chemistry ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Retina ; drug effects ; pathology

BACKGROUNDRevision anterior cruciate ligament (ACL) surgery can be expected to become more common as the number of primary reconstruction keeps increasing. This study aims to investigate the factors causing instability after primary ACL reconstruction, which may provide an essential scientific base to prevent surgical failure.

METHODSOne hundred and ten revision ACL surgeries were performed at our institute between November 2001 and July 2012. There were 74 men and 36 women, and the mean age at the time of revision was 27.6 years (range 16 - 56 years). The factors leading to instability after primary ACL reconstruction were retrospectively reviewed.

RESULTSFifty-one knees failed because of bone tunnel malposition, with too anterior femoral tunnels (20 knees), posterior wall blowout (1 knee), vertical femoral tunnels (7 knees), too posterior tibial tunnels (12 knees), and too anterior tibial tunnels (10 knees). There was another knee performed with open surgery, where the femoral tunnel was drilled through the medial condyle and the tibial tunnel was too anterior. Five knees were found with malposition of the fixation. One knee with allograft was suspected of rejection and a second surgery had been made to take out the graft. Three knees met recurrent instability after postoperative infection. The other factors included traumatic (48 knees) and unidentified (12 knees).

CONCLUSIONTechnical errors were the main factors leading to instability after primary ACL reconstructions, while attention should also be paid to the risk factors of re-injury and failure of graft incorporation.

Adolescent ; Adult ; Anterior Cruciate Ligament Reconstruction ; adverse effects ; Female ; Humans ; Joint Instability ; etiology ; Male ; Middle Aged ; Retrospective Studies

Objective Hepatitis B virus covalently closed circular DNA in the serum of patients with hepatitis B, peripheral mononuclear cells (PBMC) and liver tissue distribution. Methods Serum HBV DNA105 copies/ml 50 cases of hepatitis B patients, serum HBV DNA105 copies/hal 30 cases of hepatitis B patients, patients with fatty liver (hepatitis B) 20 cases, using real-time quantitative palymerase chain reaction for detection of serum, and liver tissue in PBMC HBVcccDNA the existence. Results Of 50 serum HBV DNA105 copies/ml cccDNA the serum specimens were 28 cases, 56% detection rate, 29 cases were PBMC HBVcccDNA, 58% detection rate, liver tissue HBVcccDNA were 44 cases, 88% detection rate, serum, the PBMC were detected in liver tissue were significant differences in P＜0.005, compared serum PBMC were no significant differences in P ＞0.005. 30 serum HBV DNA105copies/ml the serum specimens, PBMC cccDNA detected two cases were 6.67% detection rate, liver tissue cccDNA were six cases 20% detection rate, serum, PBMC, were among the liver tissue was not significantly different P＞0.005. 20 in the serum of patients with fatty liver, liver tissue in PBMCwere not detected HBVcccDNA. Conclusion HBVcccDNA mainly in the liver of patients with hepatitis B, hepatitis B patients and also cccDNA PBMC in the presence of liver tissue but a few of many.

OBJECTIVETo establish a real-time RT-PCR based plasma virus quantification method and monitor the plasma viral load of SHIV-CN97001 during its in vivo passages in rhesus macaques.

METHODSViral RNA standards were prepared by in vitro transcription and one-tube real-time RT-PCR were established and optimized using TaqMan EZ RT-PCR CORE REAGENTS and TaqMan probes and primers directed to the 91 bases within the conserved gag region of SHIV. Plasma viral RNA of 126 plasma samples from rhesus macaques of different viral passages was quantified.

RESULTSThe PCR system was optimized by using serial dilution of standards, and the viral RNA load was detected. The lowest limit of the standard curve reached 2x10(-2) copies/ml. The correlation (r>0.99) and the repetition (CV=4.14 percent) also met the requirement. It was revealed that the viral RNA load of third passage was the highest. Generally, the viral load peaks (10(5)-10(6) copies/ml) appeared at the fourteenth day after the infection or inoculation.

CONCLUSIONThe method of one-tube real-time RT-PCR was established successfully, which may provide a sensitive way to qualify SHIV viral load. This will contribute to the establishment and application of SHIV/rhesus macaque models. It was also found that the replicative ability of SHIV-CN97001 was enhanced during the first 2 in vivo passages.

Animals ; HIV ; genetics ; isolation & purification ; HIV Infections ; virology ; Humans ; Macaca mulatta ; RNA, Viral ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Serial Passage ; Simian Acquired Immunodeficiency Syndrome ; virology ; Simian Immunodeficiency Virus ; genetics ; isolation & purification ; Viral Load

OBJECTIVETo investigate the feasibility and safety of the treatment for thoracolumbar fractures with pedicle screw at the fracture level and vertebroplasty via paraspinal approach.

METHODSFrom August 2007 to August 2010, 22 old patients with thoracolumbar fractures were treated with pedicle screw at the fracture level and vertebroplasty via paraspinal approach. There were 14 males and 8 females, ranging in age from 60 to 71 years (mean, 64.6 years). The time from injury to surgery varied from 1 to 4 d (mean,2.7 d). All the patients suffered from single thoracolumbar fractures and located at T11 in 2 cases, at T12 in 5 cases, at L1 in 11 cases and at L2 in 4 cases. According to the Denis fracture classification, there were 6 compression fractures and 16 burst fractures. The mean preoperative load-sharing classification of spine fractures was 5.4 score. The mean preoperative thoracolumbar injury classification and scoring was 5.2. Based on the ASIA neurologic grading system, preoperative neurological function was grade B in 2 cases,grade C in 3 cases, grade D in 7 cases and grade E in 10 cases. The neurological function, vertebral central and anterior height, kyphotic angle of the vertebral fractures by radiographs and visual analog scale were calculated pre-operatively, post-operatively and at the last follow-up.

RESULTSMedian operating time was 60.8 min (ranged from 50 to 95 min) and median blood loss was 84 ml (ranged from 50 to 130 ml). The operative incisions were healed well. The duration of follow-up averaged 21.6 months (ranged from 12 to 48 months). The anterior vertebral body height was corrected from preoperative (52.3 +/- 10.3) % to postoperative (6.1 +/- 4.2) % and (6.8 +/- 5.4) % at the last follow-up. The central vertebral body height was corrected from preoperative (38.9 +/- 11.2) % to postoperative (8.3 +/- 4.7) % and (9.4 +/- 4.5)% at the last follow-up. The Cobbs angle of the injured vertebral segment was corrected from preoperative (19.5 +/- 9.5) degrees to postoperative (4.3 +/- 4.1) degrees and (6.2 +/- 4.7) degrees at the last follow-up. The VAS scores reduced from preoperative 8.56 +/- 0.88 to post-operative 3.48 +/- 0.91 and 3.20 +/- 0.92 at the last follow-up. The postoperative neurologic function of all 22 patients improved 1 to 2 degrees except 10 patients of grade E. There were no instances of instrumentation failure and no patient had persistent postoperative back pain.

CONCLUSIONThe pedicle screw at the fracture level and vertebroplasty via paraspinal approach has the advantages of less invasive and blood loss, and could prevent the development of kyphosis and offers improvement of the spinal cord function. Furthermore, it could decrease the risks of postoperative back pain and the failure of instrumentation.

Aged ; Bone Screws ; Female ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; injuries ; surgery ; Male ; Middle Aged ; Recovery of Function ; Retrospective Studies ; Spinal Cord ; physiopathology ; Spinal Fractures ; diagnostic imaging ; physiopathology ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Vertebroplasty ; instrumentation