Objective To investigate the clinical significance of NKp44+NK cells in the peripheral blood and synovial fluid of rheumatoid arthritis (RA) patients.Methods The proportions of NKp44+NK cells in the PB of 20 active and 15 remission patients with RA and 20 healthy individuals were detected using flow cytometry.The proportions of NKp44+NK cells in the SF of 10 active patients were detected.Multiple nonparametric test was used to compared the difference of NKp44+NK cells proportions.Clinical data including DAS28,anti-CCP antibody and RF were collected.The relationship between NKp44+NK cells and clinical data was analyzed by Spearman correlation analysis.Results The proportions of NKp44+NK cells in PB of active and remission patients and healthy individuals were (1.480±4.750)%,(0.540±0.590)%,(0.000±0.000)%respectively.The proportion of NKp44+NK cells in PB of active patients was significantly higher than that of remission patients and healthy individuals (x2=46.708,P=0.000).The proportion of NKp44+NK cells in the SF of ten active RA patients was significantly higher than matched PB.There was positive correlation between NKp44+NK cells proportion in PB and SF of active patients and DAS 28 and anti-CCP antibody level.There was no correlation between NKp44+NK cells proportion in PB,SF and RF.Conclusion There is a marked increase in the proportion of NKp44+NK cells in PB and SF of RA patients.Moreover,NKp44+NK cells are positively correlated with DAS28 and anti-CCP antibody,suggesting that NKp44+NK cells may be correlated with disease activity and severity.

BACKGROUND: Zero-P anterior cervical fixation system is an important means for treatment of cervical myelopathy in recent years. Zero-P fixation system can achieve effective decompression of spinal cord and nerve, and restore cervical curvature, with good stability.OBJECTIVE: To investigate the biocompatibility of Zero-P for multilevel cervical myelopathy.METHODS: Totally 62 patients with multilevel cervical disease who were treated by anterior cervical discectomy and fusion with Zero-P were selected, including double segments in 47 cases, three segments in 13 cases and four segments in 2 cases. Clinical efficacy was evaluated by pain visual analogue scale score, Japanese Orthopedic Association score, neck disability index score at postoperative 1, 6, 12, 24 months. Simultaneously, incidence of postoperative complications, cervical intervertebral space height and Cobb angle changes were observed.RESULTS AND CONCLUSION: (1) 54 patients were followed 2 years after treatment by clinic or telephone. Mean operative time was (102.00±32.41) minutes; average blood loss was (62.45±18.36) mL. (2) Two patients affected mild throat discomfort at one day after operation, and the symptoms went to lift six days after. The remaining patients did not experience any other complications. (3) Visual analogue scale score and neck disability index score at 1, 6, 12 and 24 months after operation were all less than preoperatively (P < 0.05); Japanese Orthopedic Association scores were higher than preoperatively (P < 0.05); the Cobb angle was improved significantly and greater than before treatment (P < 0.05).Postoperative cervical intervertebral space height was significantly higher than preoperatively (P < 0.05). Japanese Orthopedic Association results showed that among 54 patients, excellent was found in 34 cases, good in 14 cases,average in 6 cases and poor in 0 cases; the excellent and good rate was 89%. (4) Zero-P has a good clinical efficacy for multilevel cervical myelopathy, relieves symptoms significantly and restores cervical curvature and height, and has a good biocompatibility.

Objective To investigate the expression of serum soluble cytotoxic T lymphocyte associated antigen 4 (sCTLA4), the association of sCTLA4 level with erythrocyte sedimentation rate (ESR) and C reactive protein (CRP), as well as its role in patients with Crohn's disease (CD). The relationship-1661A/G and -1722T/C polymorphisms of CTLA4 gene and between disease susceptibility and phenotype of CD was analyzed. Methods A total of 126 CD patients and 300 healthy controls were enrolled in the study. Serum sCTLA4 level was determined by enzyme-linked immunosorbent assay. The concentrations of ESR and CRP were analyzed by automatic ESR Analyzer SRS 100/Ⅱ and rate nephelometry, respectively. The polymorphisms of CTLA4-1661A/G and -1722 T/C were genotyped by DNA sequencing. Results Serum sCTLA4 level was higher in CD patients than in healthy controls [(18. 70±3. 72) ng/ml vs (1.72±0. 32) ng/ml, P＜0. 01)]. Among CD patients, sCTLA4 level was higher in patients with active disease when compared to those with inactive disease [(19.83±4.35) ng/ml vs (18. 02±3.14) ng/ml, P=0. 015)]. sCTLA4 level was positively correlated with ESR and CRP levels (r=0. 267, P=0. 003; r=0. 524 P ＜0.01, respectively). In CD patients, serum sCTLA4 level was significantly higher in those with stricturing disease behavior than that in those without stricturing and penetrating or with penetrating disease behavior (P= 0.021; P=0. 015, respectively). Detection of CTLA4 -1661A/G and -1722T/C polymorphisms showed no significant difference between CD patients and healthy controls. Conclusions The high level of serum sCTLA4 in CD patients is correlated with disease activity, CRP levels and disease behavior. It suggests that sCTLA4 may play an important role in pathogenesis of CD.

Objective To investigate the function of major histocompatibility complex (MHC)class Ⅰ-related gene (MIC) in ulcerative colitis (UC) pathogenesis. Methods The difference of MICA, M ICB and their ligand NKG2D genes expression in colonic mucosa tissue of 34 UC patients and 12 healthy people were determined by fluorescent real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and the expression location of M ICA in colonic mucosa tissue was obtained by laser scanning confocal microscope. Results The mRNA level of MICA, MICB and NKG2D expression in UC groups (3. 5408±2. 6658, 8. 9879±3. 2893 and 2. 4395±0. 8147 accordingly) was significantly higher than that in the healthy controls ( 1. 0477 ± 0. 7201, 4. 6293 ± 1. 2616 and 1. 1624±0. 3954 accordingly) (P = 0.0053, 0.0039 and 0. 0078 accordingly). It suggested that MICA was expressed in colonic epithelia cell membranes by laser scanning confocal microscopy.Conclusion The mRNA level of MICA, M ICB, and their ligand, NKG2D expression were all up regulated in the colonic mucosa of UC patients, which indicated MIC gene might perform important local function in UC.

Objective: To study change of serum high sensitive cardiac troponin T (hscTnT) concentration in coronary circulation of patients with non ST-elevation acute coronary syndrome (NSTE-ACS). Methods: The subjects were all selected from our hospital, including 46 NSTE-ACS patients （NSTE-ACS group）, 42 patients with stable angina pectoris (SAP group) and 30 cases with negative coronary angiography results（healthy control group）The hscTnT concentrations in coronary venous sinus, coronary artery (CA) and peripheral serum were measured in three groups respectively. The results were compared and analyzed. Results: Compared with healthy control group, the hscTnT concentrations in coronary venous sinus, CA and peripheral venous serum all significantly increased in NSTE-ACS group and SAP group, P<0.01 all. Compared with hscTnT levels in CA and peripheral venous serum, there was significant increase in coronary venous sinus [(0.9657±0.5863) μg/L vs. (0.9562±0.7853) μg/L vs. (1.3018±1.1024) μg/L, P<0.05] in NSTE-ACS group. Conclusion: The serum hscTnT concentrations in peripheral vein, coronary artery and coronary venous sinus all significantly increase in NSTE-ACS patients, especially in coronary venous sinus.

In this study, the vaporization ratio of the 2-mum laser in the prostatic tissue with benign prostatic hyperplasia was examined in vitro, to explore a technique to estimate the clearance rate of prostatic tissue during the transurethral vaporesection of the prostate. A total of 9 fresh prostatic tissue specimens were obtained by open surgery and the wet weight of the prostatic tissue were measured immediately after the sample collection. Under the simulated conditions of transurethral vaporesection of the prostate by 2-mum laser, each prostate gland was completely vaporesected into fragments with a diameter of less than 1.0 cm in vitro. After the vaporesection, the whole fragments of prostatic tissue were collected and measured. Then the lost weight of prostatic tissue, the weight of the collected prostatic tissue and the ratio of the lost weight of prostatic tissue to the wet weight of the prostate glandular organ specimen were calculated. The correlation between the weight of collected prostatic tissue and the weight of the whole glandular organ was analyzed. All the experimental procedures were carried out by one operator. Wet weight of the prostatic gland specimen and the weight of the harvested prostatic tissues after the procedure were recorded. With respect to the wet weight of prostate gland specimen, the percentage of the weight of collected prostatic tissue was (34.45+/-1.51) %, and the percentage of the lost weight of prostatic tissue was (65.55+/-1.51)%. Satisfactory linear relationship was observed between the weight of collected prostatic tissue and the wet weight of prostate gland specimen [y=3.245x-6.475 (t=15.097, P=0.000)]. It is concluded that under the simulated conditions of transurethral vaporesection of the prostate by 2-mum laser, the vaporization ratio of prostatic tissue can be calculated on the basis of the weight of collected prostatic tissue, and thereby the clearance of prostatic tissue during the formal operation by 2-mum laser could be quantitatively determined.

BACKGROUND: Bushen Huoxue Tongluo Capsule, a well-known traditional prescription, exhibits remarkable treatment outcomes in knee osteoarthritis, but the underlying mechanism is not fully understood. OBJECTIVE: To explore the regulatory effect of Bushen Huoxue Tongluo Capsule on miR-27a of synovial cells in Sprague-Dawley rats with knee osteoarthritis. METHODS: The model of knee osteoarthritis was established in 120 Sprague-Dawley rats using Hulth method, and then these rat models were randomized into four groups, followed by given75 mg/kg normal saline (blank control group), 34 (low-dose group), 75 (medium-dose group) and 140 (high-dose group) mg/kg Bushen Huoxue Tongluo Capsule aqueous solvent via gavage according to body mass, respectively, twice daily, for consecutive 2 courses (15 days as a course) with 2 days in between. The synovium and cartilages were removed after each course, and the expression of miR-27a in the knee joint synovium and chondrocytes was determined by real-time quantitative PCR. RESULTS AND CONCLUSION: Gross observation found that the degeneration of synovium and cartilage in the Bushen Huoxue Tongluo Capsule groups was milder than that in the blank control group. Real-time PCR revealed that there was an increasing trend in the expression of miRNA-27a in the synovium and cartilage in each group, especially in the high-dose group, and there was a significant difference between groups (P < 0.05). Our findings show that Bushen Huoxue Tongluo Capsule can suppress the reduced expression of miRNA-27a in the synovium and chondrocytes in rats with knee osteoarthritis, further alleviating knee degeneration and protecting the knee from damage.

BACKGROUND:Different anesthesia methods significantly impact mean arterial pressure, systolic and diastolic blood pressure, arterial blood pH and blood viscosity, but what kind of anesthesia method is more suitable for preparation of animal model does not reach an agreement. OBJECTIVE: To compare the effectiveness of local anesthesia and general anesthesia for preparation of rabbit intervertebral disc degeneration model in procession of percutaneous acupuncture rotary cut. METHODS:A total of 48 New Zealand white rabbits were randomly divided into the local anesthesia group and the general anesthesia group. The local anesthesia group was anesthetized with 0.5% lidocaine. The general anesthesia group was intraperitoneally injected with 3% sodium pentobarbital (30 mg/kg). In the two groups, models of intervertebral disc degeneration were established by minimally invasive percutaneous acupuncture rotary cut at L4/5, L5/6 levels. Modeling time was observed in both groups. Disc degeneration was evaluated by the gross observation, MRI detection, and histopathological examination at 4, 8, 12, and 16 weeks after modeling. RESULTS AND CONCLUSION: (1) General observation: the nucleus pulposus gradually with darker color and elasticity reduced in two groups after modeling. (2) MRI T2-weighted image results showed no significant changes in disc signal intensity at early stage, but the signal strength showed a decreasing trend with time. (3) Disc degeneration was evaluated according to Pfirrmann grading. Disc degeneration was gradually increased with time in two groups (P< 0.05), but there was no significant difference at postoperative time points in both groups (P > 0.05). (4) Masson staining showed that different degrees of annulus irregular arrangement appear in the two groups 8 and 12 weeks after modeling, but the structure was still intact. 16 weeks later, annulus disorganized, or even breakage in the two groups; no significant difference was detected between the two groups. (5) Safranin O staining showed that nucleus pulposus cells were not significantly reduced in the two groups at 4 weeks, but were significantly reduced at 16 weeks. (6) The average modeling time of local anesthesia group [(15.24±2.67) minutes] was significantly shorter than that in the general anesthesia group [(25.64±6.85) minutes] (P < 0.05). These results indicate that intervertebral disc degeneration model can be successfully established by minimally invasive percutaneous acupuncture rotary cut by using local anesthesia and general anesthesia; however, local anesthesia has a shorter operating time and simple anesthetic procedure. The efficacy was identical between the two methods.

BACKGROUND:The Herbal Compound formula Yaotu Granules for lumbar disc degeneration has achieved satisfactory efficacy, but the underlying mechanism remains unclear. OBJECTIVE: To explore the possible molecular mechanisms ofYaotu Granules for lumbar disc degeneration based on transcriptome sequencing (RNA-seq). METHODS: Ten New Zealand white rabbits were randomly divided into control and experimental groups (n=5 per group), followed by intragastric injection of 20 mL of normal saline and 20 mL of water decoction of Yaotu Granules, respectively, twice daily for consecutive 1 week. The different drug serums were prepared using serum samples obtained from the common carotid artery at 2 hours after the last injection. Human nucleus pulposus cells were cultured in DMEM containing 10% different drug serums to observe the cellular morphology in each group. The cell viability was detected by trypan blue staining. The passage 3 human nucleus pulposus cells were pretreated for 48 hours, the mRNA level was detected using RNA-seq, and the differentially expressed mRNA was screened by RNA-seq. Subsequently, the differential gene expression, Gene Ontology (GO) enrichment analysis and Pathway enrichment analysis were performed. RESULTS AND CONCLUSION: In the experimental group, there were round or fusiform cells with abundant cytoplasm and clear nuclei that showed smooth surface and complete nuclear membrane; elongated rough endoplasmic reticulum arranged regularly, and less mitochondrion, scattered lysosomes and filaments were visible in the cytoplasm. In the control group, spindle and polygonal cells were found, and the large nuclei with 2 or 3 nucleoli and slightly rough surface were observed; there were mildly dilated endoplasmic reticulum, and few mitochondria with incomplete membrane. The adherent rate, generation time and cell viability of passage 1, 2 and 3 cells in the experimental group were significantly better than those in the control group (P < 0.05). Sequencing results found that there were 464 differentially expressed genes including 143 upregulated, and 321 downregulated genes. GO analysis revealed that the differentiated genes were mainly concentrated on cell regulation and metabolism. Pathway analysis found that mainly differentiated genes focused on the metabolism related pathways. These findings suggest that the differentially expressed gene profile of Yaotu Granules for lumbar disc degeneration is obtained by RNA-seq technology.Yaotu Granules mainly upregulate extracellular matrix metabolism-related genes and downregulate polysaccharide synthesis related genes in the prevention and treatment of lumbar disc degeneration, which provides basis for further research.

ObjectiveTo evaluate the clinical outcome of new proximal femoral nail antirotation (PFNA) in treatment of subtrochanteric fractures. MethodsThere were 31 patients with traumatic subtrochanteric fractures intramedullarily fixed with PFNA. The patients included 14 males and 17 fe-males at a mean age of 54.6 years (range 34-90 years). According to Seinsheimer classification of sub-trochanteric fractures, 14 patients were with type Ⅱ fractures (including four patients with type ⅡA,seven with type ⅡB and three with type ⅡC), 12 with type Ⅲ fractures (including eight with type ⅢA and four with type ⅢB), two with type Ⅳ fractures and three with type Ⅴ fractures. Close reduction was performed under traction and C-arm fluoroscopy in 26 patients and open reduction through a mini-incision before inserting the nail in five patients due to difficult close reduction. ResultsThe operation lasted for a mean time of 72.5 minutes (range 45-120 minutes), with mean blood loss of 127.5 ml (range 100-350 ml). Of all, 27 patients (87.1%, 27/31) were followed up for a mean duration of 14.3 months (range 8-24 months), which showed fracture union in all patients, with a mean union period of 17.4 weeks (range 10-21 weeks). According to Harris Hip Rate Scale, the results were. excellent in 21 pa-tients, good in two and fair in four, with excellence rate of 85.2% (23/27). ConclusionsPFNA is an effective device for treatment of subtrochanteric fracture, with a high union rate and a low complication rate, for it has advantages of easy use and minor trauma, low bleeding and stable fixation in operation.