Objective To investigate the neutrophil extracellular traps (NETs) formation and their molecular mechanisms induced by serum amyloid A (SAA) in rheumatoid arthritis (RA).Methods Neutrophils were isolated from peripheral blood of RA and healthy volunteers.① Neutrophils were cultured in vitro,the formation of NETs was observed and their percentage was calculated.② Neutrophils were cultured in vitro,divided into six groups:control,SAA,[SAA+anti-Toll like receptro4 (TLR4)-Ab],LPS,(LPS+anti-TLR4-Ab) and anti-TLR4-Ab.Appropriate stimulation was conducted for each group.NETs formation and their percentages were investigated.The concentration of DNA in supernatant was detected by fluorescent staining.F test and t test were used for statistical analysis.Results ① The purification of isolated neutrophils was higher than 95％.The network which was collocated with the spreading neutrophils nucleus and neutrophil elastase under the microscope,was NETs.In the RA group,the formation of NETs induced by SAA was significantly more than control [(19.1±0.8) vs (7.4±0.5),t=12.30,P＜0.05].② However,after pretreated with anti-TLR4 antibody,NETs formation was significantly less than the SAA group [(5.7±0.4) vs (14.7±1.1),t=7.825,P＜0.05].Moreover,the fluorescence intensity of DNA in supernatant was significantly higher in SAA group than that of anti-TLR4-Ab pretreatment group [(18.7 ±0.7) vs (12.9±0.8),t=5.552,P＜0.05].The concentration of DNA in supernatant of SAA group was higher than that of anti-TLR4-Ab pretreatment group [(36.9±1.3) μg/ml and (16.3±0.6) μ,g/ml,t=14.41,P＜0.05].Conclusion SAA can induce the formation of NETs from neutrophils by binding to TLR4 in RA.

An analytical method based on high performance liquid chromatography tandem mass spectrometry has been developed for the simultaneous determination of 20 anti-obesity drugs ( fenfluramine, phenylpropanolamine, sibutramine, sertraline, rimonabant, bupropion, citalopram, fluoxetine, benfluorex, topiramate, zonisamide, caffeine, phenolphthalein, emodin, indapamide, bumetanide, torasemide, triamterene, orlistat, phenformin). that were extracted from various weight-loss functional foods by ethanol-acetone(7:3, V/V)and purified by primary secondary amine (PSA) and octadecyltrimethoxysilane(ODS) under ultrasonication. The analysis was carried out on HPLC-MS /MS by electrospray ionization using multiple reaction monitoring after the chromatographic separation on Waters Atlantis T3 (3 μm, 150 mm × 2. 1 mm) column. Identification was achieved by the retention time and the ion ratio, quantification was done by the external standard method. The limits of detection for the appetite suppressants were 0. 05-3. 0 mg/kg. The mean recoveries at the three spiked levels were 67 . 1%-101 . 4%, with the intra-day precision less than 10%and the inter-day precision less than 15%. The method is reliable, accurate, reproducible and suitable for the determination of the anti-obesity drugs in different weight-loss functional foods.

Objective To investigate the effect of external fixator on repairing the vascular injuries in the knee and calf.Methods From May 2004 to October 2010,208 patients (234 sides) with vascular injuries in knee and calf,treated with external fixation,were retrospectively analyzed,including 192 males (217 sides) and 16 females (17 sides) with an average age of 39.6 years (range,14-67).Blood vessel damage:the popliteal artery injury in 84 patients (86 sides),the anterior tibial artery injury in 36 (46),the posterior tibial artery injury in 41(49),the anterior tibial artery and the posterior tibial artery injury in 47 (53).Complete ischemic of the injured distal limb existed in 129 patients(141 sides),incomplete ischemia in 79 patiets (93 sides).All patients underwent emergency surgery.The AO external fixators were used in 164 patients,the chuangsheng inlaid external fixators in 37,and the hybrid external fixators in 7.The injured vessels were anastomosed directly,or indirectly with great saphenous vein.The wounds were treated with phase Ⅰ bandage,skin grafts,flap transplantation or VSD suction.Results One hundred and ninty four patients obtained successful limb salvage; but 14 patients underwent amputation owing to serious damage of limb and poor general condition.One hundred and twenty seven patients underwent the second stage debridement combined with skin graft or flap transplantation; 24 patients underwent the first stage free skin graft or flap transplantation;wound cicatrized by oneself in 43 patients.One hundred and forty eight patients were followed up for 8 months to 3 years.External fixators were removed after an average of 7.2 months (range,1-18).The average healing time of fracture in 102 patients was 5.6 months (range,3-13).Forty patients suffered bone nonunion or bone defect; after treated by fracture fixation,bone graft or bone transport,all of them got bone union.Conclusion The external fixation can shorten operation time as well as the time of limb ischemia,which is an effective option on repairing vascular injuries in knee and calf.

The results of RT-PCR and immunohistochemistry showed that the expressions of vascular endothelial growth factor (VEGF) and its receptor ( flk-1 ) in the renal cortex of insulin-resistant rats during the phase of normal blood glucose were significantly increased, which were decreased by telmisartan. The result suggests that telmisartan may ease kidney damage via decreasing VEGF and flk-1 expressions.

AIM: To set up and evaluate a computerized image scoring system for quantification of posterior capsular opacification (PCO) that is independent of examinee.METHODS: The PCO model was made in rabbits. Following dilation of the pupil (＞6.5mm), standardized digital images of the PCO were obtained using slit-lamp photographic system(digital camera, computer included). PCO was scored by evaluating retroillumination photograghs. The PCO score was calculated by the formula: PCO score=∑ (OD×AI in CR)×2 +∑(OD×AI in OR), in which OD is opacity density (class 0 to 4),AI is the opacity area fraction, CR is the central region (diameter=3mm), OR is the outer region (3-6mm ring region).To evaluate the reliability of the scoring system, 6 examiners scored the pictures of 5 same eyes (interindividual reliability),and one examiner scored the pictures of 5 eyes on five different days (intraindividual reliability).RESULTS: The PCO scoring system was very reliable. With PCO score range from 0 to 1.5, the interindividual reliability showed standard deviation between 0.05-0.087. Kruskal-Wallis analysis showed no significant difference (H=0.314,P=0.9726). The intraindividual reliability showed standard deviation between 0.041-0.067, no significant difference either (H =0.613, P =0.9616).CONCLUSION: This PCO quantification analysis system evaluates the central area (diameter=6mm) of posterior capsule which is larger than visual acuity or other objective testing. The system revealed high reliability and insignificant investigator-dependent difference. With a standardized photographic setup, systematic errors by the photographic technique were not relevant. This system was proved to be an easy and useful tool to accurately evaluate PCO.

AIM:To observe the effect of simvastatin on myocardial tissue after renal ischemia-reperfusion in-jury and its mechanism .METHODS:A rat model of renal ischemia-reperfusion injury was prepared by clamping the bilat-eral renal arteries for 45 min.The rats (n=36) were randomly divided into sham operation group , renal ischemia-reperfu-sion (I/R) group and simvastatin group with 12 rats in each group.The content of serum creatinine (SCr), blood urea ni-trogen ( BUN) and myocardial tissue malondialdehyde ( MDA) , the myocardial activity of lactate dehydrogenase ( LDH) , creatine kinase (CK) and superoxide dismutase (SOD), and the myocardial protein expression of Bcl-2 and Bax were de-tected.RESULTS:Compared with sham operation group , the content of SCr , BUN and myocardial MDA , and the myo-cardial activity of LDH and CK in I/R group were significantly increased (P<0.05), and the activity of SOD was signifi-cantly decreased (P<0.05).Compared with I/R group, the content of SCr, BUN and myocardial MDA, and the myocar-dial activity of LDH and CK in simvastatin group were significantly decreased ( P<0.05 ) , while SOD activity was en-hanced (P<0.05).The protein expression of Bcl-2 and Bax in sham operation group was less than that in I/R group (P<0.05), and the protein level of Bax in simvastatin group was significantly lower than that in I /R group (P<0.05), while the protein level of Bcl-2 was increased (P<0.05).CONCLUSION:Simvastatin has a protective effect on the my-ocardium of the rats with renal ischemia-reperfusion injury , and the protective mechanism may be related to the elimination of free radicals by simvastatin , increase in the protein expression of Bcl-2 and decrease in the protein expression of Bax .

ABSTRACT:The virulence genes ,hemolytic and antibiotic resistance characteristics from 19 isolated Streptococcus suis se‐rotype 2 in Anhui Province were investigated in this study .The PCR method was developed for detecting cps2J ,mrp ,ep f and sly gene;the plate method and micro‐ELISA were developed for detecting the hemolytic type and hemolytic titer ;the K‐B method was developed for detecting antibiotic resistance of 25 kinds of antibiotics .Results showed that there were 11 strains with the dominant virulence genotype of cps2J+ /mrp+ /ep f+ /sly+ accounting for 57 .9% .All strains were α‐hemolysis orβ‐hemolysis ,with hemolytic titer of 1∶4 to 1∶128 .Strains sensitive to rifapentine ,ceftazidime ,florfenicol and cefazolin was existed in high frequency with the sensitive rates for 84 .2% ,and the resistance to doxycycline ,tetracycline ,bacitracin and sul‐famethoxazole was existed in high frequency with the resistance rates for 82 .9% .The multi‐antibiogram typing was doxycyc‐line ,tetracycline ,bacitracin and sulfamethoxazole account for 63 .2% .In conclusion ,the distribution of virulence genes of S .suis 2 Anhui isolates was similar with that of domestic reported strains ,and there was some differences with that of over‐seas reported strains .CPS ,MRP ,EPF and SLY are important virulence factors and the incompleteness of sly gene had no effect to its haemolyticus .The multi‐antibiotic resistance of S .suis 2 Anhui isolates was serious .There is difference for anti‐biogram typing between S .suis 2 Anhui isolates and those in other areas .

OBJECTIVETo explore the mechanism of action of Longbixiao Capsule (LBXC, a Chinese herbal preparation) on human prostatic stromal cell cultured in vitro.

METHODSNine Japan rabbits were assigned to 3 groups. The high, low dose group was given LBXC [2.0 g/(kg x d), 1.0 g/(kg x d)] by gastro gavage respectively, while equal volume of normal saline was given by gastrog avage 60 rats in the control group, all twice a day with an infeval of 12 h, for 3 successive days. The serum collected at 3 h after the last gastro gavage was added into cell culture fluid. Rabbit's serum containing LBXC was incubated with the cultured stromal cells, and the levels of cell proliferation and apoptosis were determined using relative techniques as TUNEL, ELISA, and immunocytochemistry. Besides, Real-time RT-PCR was applied to detect the mRNA expressions of TGF-beta1, and Smad7 in the stromal cells.

RESULTSThe cell proliferation showed culture time dependence in all groups. The proliferation in the drug-serum treated groups was lower than that in the control group, and it was lower in the high dose treated group than in the low dose treated group (all P < 0.01). The unfavorable growth did not occur morphologically after being cultured for 48 h and showed insignificant difference between various groups. Cell apoptosis was not found excepting for a few appeared in the high dose treated group (with a little amount of apoptotic cells occurring). After treatment, the expressions of TGF-beta, and Smad7 were lower in the low dose treated group and high dose treated group than in the control group (P < 0.01). There was not statistical difference between the low dose treated group and high dose treated group (P > 0.05).

CONCLUSIONLBXC could reduce the expressions of TGF-beta1, and smad7 mRNA in stromal cells and inhibit the stromal cell proliferation, but its effect on promoting cell apoptosis is unobvious.

Animals ; Apoptosis ; drug effects ; Capsules ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate ; cytology ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Random Allocation ; Serum ; Smad7 Protein ; genetics ; metabolism ; Stromal Cells ; cytology ; Transforming Growth Factor beta1 ; genetics ; metabolism

Altitude ; Exercise/physiology* ; Heart Rate/physiology* ; Humans ; Lung ; Respiration

Objective:To analyze the characteristics of nerve injury in patients with diabetic peripheral neuropathy (DPN) and explore the diagnostic value of current perception threshold (CPT) and nerve conduction velocity (NCV) for DPN.Methods:One hundred and thirty-six DPN patients admitted to Beijing Puren Hospital from June 2017 to December 2019 were selected, and 130 diabetic non-DPN patients admitted during the same period were used as controls. All the subjects were tested by CPT and NCV. Among them, the detection of NCV included sensory nerve conduction velocity (SCV) and motor nerve conduction velocity (MCV). Statistical analysis was performed on the relevant indicators of the two groups of subjects.Results:The CPT values of the upper limb median nerve and ulnar nerve at 2 000 Hz, 250 Hz, and 5 Hz, the superficial peroneal nerves at 250 Hz, and 5 Hz, and the CPT values of the sural nerve at 2 000 Hz and 250 Hz were higher than those of non-DPN patients, DPN injury mainly occurred in myelinated nerve fibers in the lower extremities [60.29%(82/136)], and the difference were statistically significant ( P<0.05). The SCV and MCV of the median nerve, ulnar nerve, and common peroneal nerve in the DPN group were lower than those in the non-DPN group, the SCV abnormal rate was higher than the MCV: 55.88%(76/136) vs. 37.50%(51/136); 58.82%(80/136) vs. 41.18% (56/136); 67.65%(92/136) vs. 50.00%(68/136), and the differences were statistically significant ( P<0.05). The area under the predictive value curve (AUC) of CPT for DPN was 0.815 (95% CI 0.735 ~ 0.895). The AUC of NCV for DPN was 0.875 (95% CI 0.813 ~ 0.944). The AUC of CPT and NCV for DPN was 0.923 (95% CI 0.876 ~ 0.970). Conclusions:DPT patients have abnormal CPT and NCV, and nerve damage occurs mostly in myelinated nerve fibers and SCV. Diagnosing DPN by combining CPT and NCV is helpful to improve the detection rate of DPN.