Objective:To explore the changes of NK and NKT cells and the expression levels of their activated, inhibitory receptors in the peripheral blood of patients with newly diagnosed rheumatoid arthritis ( RA), and to reveal the potiential role of NK and NKT cells played in the pathogenesis of RA.Methods:32 patients with new onset RA and 15 healthy controls were recruited.Activated and inhibitory NK and NKT cells in peripheral blood were quantified by flow cytometry.The frequency of spontaneous and stimulated IFN-γ+NK and NKT cells and CD107aγ+NK cells were examined.Finally,the potential relationship between the frequency of NK and NKT cells subsets and clinical indexes were analyzed.Results:The frequencies of NK cells in peripheral blood in RA patients were sig-nificantly lower than those in the controls ( P=0.026 ).The frequencies of NKG2D+, NKP46+activated NK cells and NKG2C+, NKG2D+,NKP46+activated NKT cells in RA patients were significantly higher than those in the controls (P=0.011,P=0.010,and P<0.001,P=0.032,P=0.001,respectively),whereas the frequencies of KIR2DL3+,KIR3DL1+and NKG2A+inhibitory NK cells and KIR2DL3+,NKG2A+inhibitory NKT cells in RA patients were significantly lower (P=0.002,P=0.002,P=0.014,and P=0.027,P=0.002,respectively).Moreover, the frequencies of stimulated IFN-γ+NK cells and IFN-γ+NKT cells, spontaneous and stimulated CD107 aγ+NK cells in RA patients were significantly higher than that in the controls ( P=0.037, P=0.004 and P=0.001, P=0.001, respectively).Furthermore,the frequencies of NK cells,NKG2Aγ+and KIR2DL3γ+inhibitory NK cells were correlated significantly with the values of DAS28 in RA patients (r=0.357,P=0.045;r=0.399,P=0.024;r=0.468,P=0.021,respectively).Conclusion:Lower frequencies of NK cells, higher frequencies of activated NK cells and activated NKT cells, lower frequencies of inhibitory NK cells and inhibitory NKT cells, and higher NK cell activity may induce the autoimmune reaction involved in the pathogenesis of RA.

Microscopy, Electron, Transmission ; methods ; Negative Staining ; methods ; Viruses ; ultrastructure

OBJECTIVETo explore the mechanism of pricking blood therapy (PBT) for allergic rhinitis (AR).

METHODSThe model rat of experimental allergic rhinitis (EAR) was established by administration of ovoglohulin and they were divided into five groups randomly: a model group, a PBT group, a TCM point application group, a beclomethasone dipropionate (BDP) treatment group, and a health control group. Adrenocorticotrophic hormone (ACTH) contents in hypothalamus, pituitary gland, adrenal gland and plasma were determined by radioimmunoassay before and after treatment.

RESULTSThe ACTH levels in the hypothalamus, pituitary gland, adrenal gland and plasma in the rat of EAR were lower than those in the health control group (P < 0.01). After treatment of pricking blood ther, ACTH levels in the above tissues increased significantly (P < 0.01), with very significant differences (P < 0.01) as compared with the model group, similar to those in the BDP group (P > 0.05).

CONCLUSIONPricking blood therapy has a regulatory action on ACTH in HPA axis of rats with experimental allergic rhinitis.

This study investigated the effects of concentration, intestinal section, pH and P-gp on the absorption of daphnetin. The absorptions of three concentrations (10, 20, 40 microg x mL(-1)) of daphnetin in different intestinal segments were studied with phenol red as the marker by in situ rats single pass perfusion model. The results showed that daphnetin was stable under pH 6.0 condition and little affected by metabolism enzyme. There was upgrade tendency between the Peff of duodenum, jejunum, ileum and colon in different concentration of daphnetin, and it has obvious difference between the high concentration and low concentration in jejunum and colon, which indicated that the absorption of daphnetin was passive diffusion and no difference in different segments of rat intestine. However, compared with colon, the absorption of small intestine was better significantly (P < 0.05). Daphnetin may be not a substrate of P-gp as verapamil had not significantly affected the absorption of daphnetin in different intestinal segments of rats.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Female ; Hydrogen-Ion Concentration ; Intestinal Absorption ; drug effects ; Male ; Perfusion ; Permeability ; Rats ; Rats, Sprague-Dawley ; Umbelliferones ; pharmacokinetics

Objective To explore the metabolic abnormality in patients with first-episode schizophrenia and effect of atpical antipsychotics olanzapine on them.Methods Thirty patients with first-episode schizophrenia,admitted to our hospital from February 2010 to February 2011,and 40healthy controls were chosen in our study; patient group was given oral olanzapine for 4 week.The height,weight,waistline and hipline were measured before and after treatment,and the total cholesterol (TC),triglyceride(TG),high-density lipoprotein(HDL),low-density lipoprotein(LDL),apolipoprotein AI(aPOAI),apolipoprotein B100(aPOB100),lipoprotein a(LPa),fasting blood-glucose,fasting insulin and C peptide levels were detected before and after treatment; and the insulin resistance(IR)index,waist hip ratio(WHR)and body mass index(BMI)were calculated before and after treatment; metabolic index were compared between patients and controls,and patients before and after the treatment.Results The HDL and aPOA1 levels in patient group were significantly lower,and the WHR,IR index,fasting insulin and C peptide levels were obviously higher as compared with those in the controls(P＜0.05).The BMI,waist circumference,WHR,the IR index,the insulin,TC,TG,LDL and aPOB100 levels after treatment were significantly increased as compared with those before treatment(P＜0.05).Conclusion There may be some inborn metabolic abnormality factors in patients with first-episode schizophrenia; the high incidence of metabolic abnormality in patients with schizophrenia may be the result of combined action from antipsychotics and inherit susceptibility.

A patient referred to our hospital, diagnosed with left idiopathic chronic orchialgia, was evaluated with a thorough medical and psychiatric history, physical examination, scrotal ultrasound and magnetic resonance imaging. Conservative management failed. The patient had temporary pain relief after undergoing outpatient cord block three times. Microsurgical denervation of the left spermatic cord was operated in March, 2011. A pain questionnaire was used to determine efficacy before and after operation, and complete pain relief was noted at one week after operation. The follow up period was 12 months, at the end of which the pain score was still zero. No complications, including testicular atrophy and hydrocele, occurred. Microsurgical denervation of the spermatic cord can be a minimally invasive, safe and effective management option for treatment of idiopathic chronic orchialgia.
Denervation ; methods ; Humans ; Male ; Middle Aged ; Spermatic Cord ; surgery ; Testicular Diseases ; surgery

Objective:To compare the clinical results of modified posterolateral approach combined medial approach versus traditional posterolateral approach combined with medial approach in open reduction and internal fixation of trimalleolar ankle fracture.Methods:A retrospective cohort study was used to analyze the clinical data of 46 patients with trimalleolar ankle fracture admitted to the First Affiliated Hospital of Naval Military Medical University from June 2013 to June 2019, including 14 males and 32 females, at age of 19-71 years [(49.2±14.9)years]. There were 33 patients with supination-external rotation type IV ankle fracture and 13 with pronation-external rotation type IV ankle fracture according to Lauge-Hansen classification. Open reduction and internal fixation was performed through the modified posterolateral approach combined with medial approach in 25 patients (modified approach group), and through the traditional posterolateral approach combined with medial approach in 21 patients (traditional approach group). The visual analogue score (VAS) at 3 days and 1 week after surgery, fracture healing time, range of ankle flexion and extension and Baird-Jackson score at the final follow-up and postoperative complications (numbness of the affect limb, wound necrosis, etc.) were compared between the two groups.Results:All patients were followed up for 11-21 months [(14.8±2.2)months]. There was no statistical difference in VAS or fracture healing time at 3 days after surgery between the two groups (all P>0.05). The VAS was 3.0 (3.0, 4.0)points in modified approach group at 1 week after surgery, significantly lower than 4.0 (3.0, 5.0)points in traditional approach group ( P<0.05). At the final follow-up, there was no statistical difference in range of ankle plantarflexion between the two groups ( P>0.05), but range of ankle dorsiflexion was significantly greater in modified approach group [(11.8±2.8)°] than that in traditional approach group [(8.1±3.5)°] ( P<0.01). At the final follow-up, Baird-Jackson score was not statistically different between the two groups ( P>0.05). There were 4 patients with numbness and 2 with wound necrosis in traditional approach group, but no numbness or wound necrosis occurred in modified approach group ( P<0.01). Conclusion:Both the modified posterolateral approach combined with medial approach and traditional posterolateral approach combined with medial approach can achieve good clinical results in open reduction and internal fixation of trimalleolar ankle fractures, but the former has advantages of better pain relief, better recovery of ankle dorsiflexion and less complications.

Objective To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.Methods A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells.The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot,respectively.Cell proliferation was evaluated by CCK-8 assay,cell cycle and apoptosis were tested by flow cytometry,and cell invasion was detected by transwell invasion assay.Results The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P ＜0.001).Western blot analysis proved that the relative expression of EGFL7 protein in NC group,Lenti-NCgroup and Lenti-EGFL7 group was (0.39 ±0.12),(0.36 ±0.14) and (0.07 ±0.04),respectively.EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P ＜ 0.001),which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells.The proliferation of Hep-2 cells was significantly inhibited after transfection (P ＜ 0.01).Compared with the NC group and Lenti-NC group,the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P ＜ 0.01),and the proportion in G1 phase was significantly decreased (P ＜ 0.05).Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) ％ was significantly increased in NC group (6.09-±3.28)％ and Lenti-NC group (9.86 ±2.13)％.In Transwell invision assay,the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P ＜ 0.01).Conclusion The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.

Apolipoprotein AI (apo AI), the major protein component of human high-density lipoprotein (HDL), is a single-chain polypeptide of 243 amino acids. Several epidemiological studies have shown that the plasma concentrations of HDL has the role of reverse cholesterol transport (RCT) and inversely correlated with the incidence of coronary artery disease. Because apo AI lacks post-translational modifications, it is convenient to express human apo AI in Escherichia coli expression system. However, there is a poor stability of the mRNA and the apo AI protein in E. coli, it is difficult to express mature apo AI in recombinant bacteria, moreover, even as a fusion protein, apo AI is still sensitive to degradation and can not be cleaved efficiently from the fusion tags. In contrast, proapolipoprotein AI (proapo AI, having an additional polypeptide containing the amino acids Arg-His-Phe-Trp-Gln-Gln at the amino-teminal of the mature protein) proved stable and undegraded in Escherichia coli, and therefore, in this research, an expression system of E. coli including a plasmid of P(R)P(L) tandem promoter was adapted to produce proapo AI. Furthermore, site-directed mutagenesis of the proapo AI cDNA was performed to generate a Clu8Asp mutation in the amino-terminal sequence of proapo AI which created an acid labile Asp-Pro peptide bond between amino acid 8 and 9, and permitted specific chemical cleavage to remove pro-peptide. After inducing with a shift of temperature, yields of recombinant proapo AI achieved about 40% of total cell protein and the recombinant proapo AI expressed proved as a form of inclusion body in cells, so protein need to renature. First of all, the protein was dissolved in buffer with denaturant, and renaturation was carried out on a hydrophobic interaction column (Phenyl Sepharose), ion-exchange chromatography and gel-filtration chromatography were then used to further purify the protein. The purified recombinant apo AI was detected by a set of tests including Western-blotting, Circular dichroism spectra and lipid-binding test, the results shown that recombinant apo AI has similar structural and lipid-binding properties identical to those of native plasma apo AI, which facilitates further research and application.
Apolipoprotein A-I ; biosynthesis ; genetics ; Chromatography, Ion Exchange ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Protein Precursors ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

Objective:To evaluate the efficacy and safety of sodium hyaluronate gel in preventing adhesion after prophylactic enterostomy.Methods:One hundred and twenty four patients from 6 hospitals were enrolled in this prospective multi-center randomized controlled trial. Patients were randomized into the study group ( n=59) or the control group ( n=65).All patients underwent prophylactic enterostomy. Patients of study group received odium hyaluronate gel for adhesion-prevention,while those in control group did not receive any adhesion-prevention treatment. The incidence of moderate to severe adhesion around the incision in the stoma area were evalutated during stoma reduction surgery. Results:The incidence of moderate to severe adhesion around the incision in the stoma area was 6.3% in the study group, the difference was statistically significant ( P<0.05) compared to that of the control group (32.6%). Conclusion:Sodium hyaluronate gel can safely and effectively reduce the incidence of moderate and severe adhesions after abdominal surgery.