OBJECTIVETo explore the effect of Bushen Gujin Recipe (BGR) on serum and synovial expression of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in knee osteoarthritis (KOA) model rabbits.

METHODSTotally 36 8-month-old healthy male New Zealand white rabbits were randomly divided into 4 groups, i.e., the normal control group, the model group, the Western medicine group (Meloxicam, at the daily dose of 6 mg/kg), and the TCM group (BGR, at the daily dose of 53 g/kg), 9 in each group. Modeling was performed in all rabbits except those in the normal control group by using Hulth A method. All medication was performed for 8 consecutive weeks. Contents of IL-1 and TNF-α were detected using ELISA from serum, partial synovial tissue of the front knee joint, cartilage and subchondral bone of the medial femoral condyle.

RESULTSThe joint space became narrowed in the Western medicine group, ranging between the model group and the TCM group. The articular surface was rough with obvious osteophytes. The joint space was slightly narrower in the TCM group; the articular surface was slightly rough with mild osteophytes. Compared with the normal control group, contents of IL-1 and TNF-α in serum and synovial increased in the model group (P < 0.01). Compared with the model group, contents of IL-1 and TNF-α in serum and the synovial fluid decreased in the two treatment groups (P < 0.01). There was no statistical difference in contents of IL-1 and TNF-α between the Western medicine group and the TCM group.

CONCLUSIONBGR promoted the synthesis of cartilage matrix and carti- lage repair through inhibiting the secretion of IL-1 and TNF-α, and prolonging cartilage degeneration.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1 ; metabolism ; Knee Joint ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rabbits ; Synovial Fluid ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

3 mg/L was significantly worse than in those with hs-CRP≤3 mg/L (18.18%,5.45%;P=0.044,log-rank test). Higher hs-CRP concentration was an independent predictor of death or new vascular event(OR 3.609;95% CI 0.869—14.992;P=0.047).Conclusion Higher hs-CRP concentration in acute phase after ischemic stroke is an independent predictor of death or new vascular event in a year.

OBJECTIVETo evaluate the short-term efficacy and safety of Bushen Shuji Granule (BSG) in treating ankylosing spondylitis (AS) patients.

METHODSA prospective randomized controlled clinical trial was carried out in 62 active stage AS patients with Shen deficiency Du-channel cold syndrome (SDDCS), who were randomly assigned to the BSG group (treated with BSG) and the control group (treated with Celecoxib Capsule). Twelve weeks consisted of one therapeutic course. Therapeutic effects were evaluated by ASAS20 and ASAS40 (set by Assessments in Ankylosing Spondylitis working group) , BASDA150, Chinese medical (CM) syndrome efficacy evaluation standards. BASDAI, the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath AS Metrology Index (BASMI), scores for spine pain, scores for pain at night, patient global assessment (PGA) , erythrocyte sedimentation rate (ESR) , and C reactive protein (CRP) were observed before and after treatment.

RESULTSAfter three-month treatment by BSG, ASAS20 standard rate was 63. 33% (19/30 cases) in the BSG group and 66.67% (20/30 cases) in the control group with no significant difference between the two groups (χ2 = 0.073, P > 0.05). The efficacy for CM syndromes was 70.00% (21/30 cases) in the BSG group, higher than that in the control group [40.00% (12/30 cases), χ2 = 5.455, P < 0.05]. Scores for CM syndromes, BASDAI, night pain index, spinal pain index, PGA, CRP were improved in the BSG group (P < 0.05, P < 0.01). The incidence of adverse events in the BSG group was lower than that of the control group.

CONCLUSIONBSG based on Shen supplementing, Du-channel strengthening, blood activating, and channels dredging method had good short-term clinical efficacy and safety in treating AS.

Asian Continental Ancestry Group ; Biomedical Research ; Blood Sedimentation ; C-Reactive Protein ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Pain ; Prospective Studies ; Safety ; Spondylitis, Ankylosing ; drug therapy

OBJECTIVETo evaluate multiplex ligation-dependent probe amplification (MLPA) assay detection in analysis of chromosome 22q11.2 microdeletion.

METHODSBetween March 2008 and September 2009, thirty-two patients including 10 males and 16 females aged between years (3.6±3.1) were selected and evaluated by history, physical examination and medical records. Of these patients, sixteen patients who were previous diagnostic as 22q11.2 microdeletion were in positive control group, the other 16 healthy children were in negative control group. All the patients were detected by MLPA and fluorescence in situ hybridization (FISH) for the presence of a 22q11.2 microdeletion after informed consent. Diagnostic efficacy was assessed by sensitivity, specificity and Kappa analysis.

RESULTSWe have applied the two assays of detection of chromosome 22q11.2 microdeletion in 32 patients. Sixteen patients in positive control group were found to have a 22q11.2 deletion and, with the deletion size of 3-Mb. However, as expected, chromosome 22q11.2 deletion was not found in negative control group. The MLPA results were in good agreement with that by FISH. Therefore, MLPA has high sensitivity and specificity.

CONCLUSIONMLPA is a rapid, reliable, high-throughput and relatively economical alternative to FISH technology for the diagnosis of 22q11.2 microdeletion. It can provide reliable and helpful information for clinical diagnosis of 22q11.2 microdeletion syndrome.

Child, Preschool ; Chromosome Deletion ; Chromosomes, Human, Pair 22 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Nucleic Acid Amplification Techniques ; methods ; Sensitivity and Specificity

Abstract：Objective To prepare human monoclonal antibody against spike protein（S protein）of severe acute respiratory syndrome coronavirus 2（SARS⁃CoV⁃2）by using single B cell，and determine its neutralizing activity. Methods Venous blood with high antibody level was collected from people immunized with inactivated SARS⁃CoV⁃2 vaccine（Vero cells） twice，of which peripheral blood mononuclear cells（PBMCs）were isolated by lymphocyte stratified fluid and used to isolate single B cell expressing S protein antibody by magnetic beads coupled with S1 protein. Variable region genes of IgG heavy chain and light chain were amplified by nested PCR after reverse transcription of single B cell，which were connected with CMV promoter，IgG leader sequence，IgG constant region and polyA sequence by overlapping PCR to construct antibody linear expression cassette. Linear expression cassette of the heavy chain and light chain from the same B cell was transfected to HEK293T cells to express human monoclonal antibody of SARS⁃CoV⁃2 S protein. Immunoreactivity was detected by immuno⁃ fluorescence while neutralizing activity by pseudovirus neutralization test. Results A total of 26 monoclonal antibodies against SARS⁃CoV⁃2 S protein were expressed，which showed heavy chain and light chain protein bands of IgG antibody at

The purpose of this study was to investigate the vasorelaxing effect of vasonatrin peptide (VNP) on human intramammary artery (HIMA).The vasorelaxing effect of VNP on HIMA was measured by means of perfusion in vitro. The effects of HS-142-1, TEA, 8-Br-cGMP and methylene blue (MB) were also observed. It was found that VNP caused a concentration-dependent relaxation in HIMA which was independent of the endothelium. 8-Br-cGMP (0.1-1000 micromol/L) also caused a concentration-dependent relaxation in HIMA. The vasorelaxing effect of VNP disappeared in the presence of HS-142-1 (20 micromol/L), an antagonist of the natriuretic peptide guanylate cyclase (GC) receptor. MB (10 micromol/L), an inhibitor of GC, not only blocked completely the relaxation of HIMA, but also enhanced the vascular contraction induced by norepinephrine. TEA (1 mmol/L), an antagonist of calcium activated potassium channels (K(Ca)), reduced but not completely blocked the vasorelaxing effect of VNP. These findings suggest that VNP can relax HIMA, which is independent of the endothelium. This effect is possibly achieved by the binding of VNP with the natriuretic peptide GC receptors in the smooth muscle cells (SMCs), leading to an increase in intracellular cGMP level. Moreover, the vasorelaxing effect of VNP is associated with K(Ca).
Aged ; Atrial Natriuretic Factor ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; In Vitro Techniques ; Mammary Arteries ; drug effects ; physiology ; Middle Aged ; Potassium Channels, Calcium-Activated ; metabolism ; Receptors, Guanylate Cyclase-Coupled ; metabolism ; Vasodilation ; drug effects ; physiology

BACKGROUNDThe pathophysiology of type 2 diabetes is progressive pancreatic beta cell failure with consequential reduced insulin secretion. Glucotoxicity results in the reduction of beta cell mass in type 2 diabetes by inducing apoptosis. Autophagy is essential for the maintenance of normal islet architecture and plays a crucial role in maintaining the intracellular insulin content by accelerating the insulin degradation rate in beta cells. Recently more attention has been paid to the effect of autophagy in type 2 diabetes. The regulatory pathway of autophagy in controlling pancreatic beta cells is still not clear. The aim of our study was to evaluate whether liraglutide can inhibit apoptosis and modulate autophagy in vitro in insulinoma cells (INS-1 cells).

METHODSINS-1 cells were incubated for 24 hours in the presence or absence of high levels of glucose, liraglutide (a long-acting human glucagon-like peptide-1 analogue), or 3-methyadenine (3-MA). Cell viability was measured using the Cell Counting Kit-8 (CCK8) viability assay. Autophagy of INS-1 cells was tested by monodansylcadaverine (MDC) staining, an autophagy fluorescent compound used for the labeling of autophagic vacuoles, and by Western blotting of microtubule-associated protein I light chain 3 (LC3), a biochemical markers of autophagic initiation.

RESULTSThe viability of INS-1 cells was reduced after treatment with high levels of glucose. The viability of INS-1 cells was reduced and apoptosis was increased when autophagy was inhibited. The viability of INS-1 cells was significantly increased by adding liraglutide to supplement high glucose level medium compared with the cells treated with high glucose levels alone.

CONCLUSIONSApoptosis and autophagy were increased in rat INS-1 cells when treated with high level of glucose, and the viability of INS-1 cells was significantly reduced by inhibiting autophagy. Liraglutide protected INS-1 cells from high glucose level-induced apoptosis that is accompanied by a significant increase of autophagy, suggesting that liraglutide plays a role in beta cell apoptosis by targeting autophagy. Thus, autophagy may be a new target for the prevention or treatment of diabetes.

Animals ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Glucagon-Like Peptide 1 ; analogs & derivatives ; pharmacology ; Glucose ; pharmacology ; Insulinoma ; pathology ; Liraglutide ; Rats

OBJECTIVETo evaluate the therapeutic effect of three-dimensional digital orthopedic techniques in treatment of acetabular fractures.

METHODSWe retrospectively analyzed 50 cases of acetabular fracture treated between March, 2007 and December, 2013. The lamellar CT scanning data were imported into Mimics software, and 3D anatomical models of the pelvic and proximal femur were reconstructed. Computer-assisted analysis was carried out to understand the condition of fractures and simulate fracture reduction. The pelvic models were manufactured by rapid prototyping technique for definite diagnosis and typing of acetabular fractures and subsequent surgical treatment.

RESULTSThree-dimensional reconstruction images and rapid prototyping pelvic models faithfully represented the findings in operations. Preoperative simulation of the operation shortened the time of operation and reduced the volume of bleeding in the operation. All the patients were followed up for 6 to 24 months. According to Matta imaging score, anatomical reduction was achieved in 41 cases and satisfactory reduction in 9 cases. According to the Harris functional criteria, 32 patients had excellent, 12 had good and 6 had acceptable outcomes with a rate of excellent and good outcomes of 88%.

CONCLUSIONThree-dimensional digital orthopedic techniques allow accurate display of the acetabulum and the spatial relation of the anatomic structures to assist in fracture diagnosis, typing and treatment.

Acetabulum ; pathology ; Femur ; Fracture Fixation, Internal ; Fractures, Bone ; Humans ; Imaging, Three-Dimensional ; Models, Anatomic ; Orthopedics ; methods ; Retrospective Studies ; Software ; Tomography, X-Ray Computed

OBJECTIVETo construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.

METHODSPIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot.

RESULTSWe successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.

CONCLUSIONThe construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.

Animals ; Cell Line ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Mice ; Plasmids ; Protein Inhibitors of Activated STAT ; genetics ; Spermatocytes ; cytology ; Transfection

BACKGROUNDPatellar fracture and cruciate ligament injury are a common consequence of traumatic knee injury. Patellar fracture combined with cruciate ligament injury is rarely reported, although the mechanisms of two things are similar. This study aimed to evaluate the incidence of closed patella fracture combined with cruciate ligament injury.

METHODSFrom 2012 March 1 to June 30, magnetic resonance images of 60 patients with unilateral closed patellar fracture were studied in our institution. The mean age of the patients at presentation was 40.2 years (range, 13-64 years) and 48 patients were men. First, patients were divided according to the cause of injury. Twenty-eight patients had high-energy trauma from a falling injury or motor vehicle accident, and 32 patients had low-energy trauma resulting from a tumbling injury. Second, according to the fracture pattern, 31 patients had a transverse fracture and 29 patients had a comminuted fracture.

RESULTSWe found seven cases of closed patellar fracture combined with cruciate ligament injury among 60 patients, including two cases of a completely ruptured posterior cruciate ligament, two with a partially torn posterior cruciate ligament, and three with a partially torn anterior cruciate ligament. The percentage of this combined injury was 11.6% (7/60). The incidence of a combined injury of the cruciate ligament with a comminuted fracture (6/29, 20.7%) was significantly higher than that with a transverse fracture (1/31, 3.2%, P < 0.05). The most common mechanism of injury in patellar fracture combined with cruciate ligament injury was high-energy trauma from road traffic accidents (94%), whereas in the patellar fracture alone, it was tumbling (62%). The incidence of combined injury with high-energy trauma (6/28, 21.4%) was significantly higher than that with low-energy trauma (1/32, 3.1%, P < 0.05).

CONCLUSIONSThese data suggest that high-energy trauma often results in a comminuted patellar fracture, which is often combined with cruciate ligament injury. Traffic accidents are the main risk factor for this combined injury. Understanding the relationship between patellar fracture and cruciate ligament injury for diagnosis and treatment is important.

Adolescent ; Adult ; Anterior Cruciate Ligament Injuries ; Case-Control Studies ; Female ; Fractures, Closed ; diagnosis ; Humans ; Knee Injuries ; diagnosis ; Knee Joint ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Patella ; injuries ; Young Adult