Objective To determine the role of the platelet activation and the expression of platelet Toll-like receptor 4 (TLR4) in thrombocytopenia induced by lipopolysaccharide (LPS) in mice.Methods ICR mice were randomly (random number) divided into control group,model 3,6,12,24,48,72h groups and neutrocytopenia syndrome (NEP) group.The blood samples of mice in model groups were detected 3,6,12,24,48,and 72 hours after intravenous injection of LPS respectively.Anti- neutrophil monoclonal antibody was administered in the mice of NEP group 24 h before injection of LPS,and blood samples were collected 24 h after injection of LPS.The determination of platelet count (PC),mean platelet volume (MPV) and platelet distribution width (PDW) was carried out with full automatic hemocyte analyzer.The plasma tumor necrosis factor- α (TNF- α),macrophage colony stimulating factor (M-CSF) and soluble CD40 ligand (sCD40L) levels were measured by using ELISA.The rate of platelet TLR4 expression was detected by flow cytometry.Comparison between groups was carried out by using ANOVA statistical methods.Results PC was reduced by 30％ 3 h after intravenous injection of LPS,and reached the lowest level within 24 h (P ＜0.01 ).MPV and PDW were increased compared with healthy controls (P ＜0.01 ).Plasma TNF - α,M - CSF and sCD40L were significantly increased after LPS stimulation (P ＜0.01 ),and reached the peak during 6 ～ 24 h after LPS administration.PC had correlation with plasma levels of M - CSF ( r =- 0.746) and sCD40L ( r =- 0.573 ).The platelet TLR4 expression was significantly increased 6 h after LPS stimulation.The platelet TLR4 expression in NEP group was significantly lower than that in 24 h model group ( P ＜ 0.01 ).Conclusions The excessive activation platelet represented by increase in sCD40L,MPV,PDW and high levels of M-CSF in macrophages along with platelet TLR4 expression participate in the pathogenesis of thrombocytopenia.The up - regulation of the platelet TLR4 expression is dependent on neutrophils in case of thrombocytopenia induced by LPS.

Objective To study the iodine deficiency disorders (IDD) situation in Wenzhou City during 1995-2014.Method According to National IDD Surveillance Project,IDD surveillance had been consecutively carried out during the past 20 years,which consisted of goiter rate in 8-10 years old children,iodized salt and urinary iodine levels.Results The goiter rate of 8-10 years old children was decreased from the highest of 31.09％ (2 190/7 043) in 1995 to the lowest of 2.28％ (77/3 378) in 2014;the highest level of median urinary iodine was 214.78 μg/L,and the lowest level was 74.48 μg/L,and which was increased from 74.48 μg/L in 1995 to 187.00 μg/L in 1996,and then had been maintained at the appropriate level recommended by World Health Organiation (WHO),except that in 1998,2003,2004 and 2006.The qualified rate of iodized salt was increased from 54.95％ (1 471/2 677) in 1996 to 95.52％ (2 548/2 754) in 1999,but decreased to 62.75％ (768/1 224) in 2003,however it was fluctuated from 78.61％ (2 503/3 184) to 92.48％ (2 989/3 232) from 2004 to 2013,and it was 90.43％ in 2014 (2 983/3 300).Conclusions The comprehensive measures for controlling IDD,with universal salt iodization,has been gradually achieved remarkable effect in Wenzhou City,but the non-iodization salt existing in the market is still a problem,and people have misunderstandings about iodization salt.Iodine supplementation had better be conducted according to local conditions and based on scientific policy.

Objective To know the establishment of the flap pedicle blood supply and the right moment of the cutting off of pedicle by means of laser Doppler blood perfusion imaging instrument on the random flap blood flow changes.Methods 20 adult healthy rabbits were divided in to four groups and each 5 have unilateral flaps.1,3,5,7,9,12,14,18 days after operation,the pedicle blood perfusion values (PU) measurement of the distal pedicle on the flap and midpoint of both ends of the pedicle were performed,and PU values were analyzed.Results Pedicle of the PU values at the different time points changed little (P＞0.05).The PU value in the distal flap reached the minimum 1 day after operation and then increased gradually [3 d (1.24±0.07),5 d (1.57±0.15),7 d (1.79±0.08),9 d(1.89±0.13),12 d(2.01±0.16),14 d(2.18±0.09) and 18 d(2.40±0.18),P＜0.05].When distal PU values/pedicle PU value≥1.2,the flap survival rate reached 99%.Conclusions The establishment of random skin flap blood circulation,as well as the ratio of PU values of distal to the pedicle flap pedicle can be used as the timing of an important indicators.

The aim of this study was to establish an assessment method for determining α-Gal (α-1, 3-galactosyle) epitopes contained in animal tissue or animal tissue-derived biological materials with ELISA inhibition assay. Firstly, a 96 well plate was coated with Gal α-1, 3-Gal/bovine serum albumin (BSA) as a solid phase antigen and meanwhile, the anti-α-Gal M86 was used to react with α-Gal antigens which contained in the test materials. Then, the residual antibodies (M86) in the supernatant of M86-Gal reaction mixture were measured using ELISA inhibition assay by the α-Gal coating plate. The inhibition curve of the ELISA inhibition assay, the R2 = 0.999, was well established. Checking using both α-Gal positive materials (rat liver tissues) and α-Gal negative materials (human placenta tissues) showed a good sensitivity and specificity. Based on the presently established method, the α-Gal expression profile of rat tissues, decellular animal tissue-derived biological materials and porcine dermal before and after decellular treatment were determined. The M86 ELISA inhibition assay method, which can quantitatively determine the α-Gal antigens contained in animal tissues or animal tissue-derived biomaterials, was refined. This M86 specific antibody based-ELISA inhibition assay established in the present study has good sensitivity and specificity, and could be a useful method for determining remnant α-1, 3Gal antigens in animal tissue-derived biomaterials.
Animals ; Antibodies ; Biocompatible Materials ; Enzyme-Linked Immunosorbent Assay ; methods ; Epitopes ; analysis ; Humans ; Rats ; Sensitivity and Specificity ; Serum Albumin, Bovine ; Trisaccharides ; analysis

BACKGROUND: Pentoxifylline (PTX), as an effective drug to improve blood rheology, has been used as a vasodilator for the treatment of vascular diseases and peripheral vascular disease science 1960s. But the role of PTX on skin blood flow to improve flap survival remains still unclear. OBJECTIVE: Through the clinical application of PTX therapy, the dynamic observation of random flap to understand that the PTX can promote flap survival and improve blood circulation of flap. METHODS: A total of 39 patients (27 males and 12 females aging 7-54 years) with skin tissue defect undergoing random flap repairing were selected from Department of Burn and Plastic Surgery, First Hospital of Hebei Medical University. The 39 cases were randomly divided into control group and drug group. Patients in the drug group were given intravenous injection of 250 mL PTX at day 2 after random flap operation, once a day, until 14 days after flap repairing surgery. On the first day after flap pedicle surgery, 250 mL PTX sodium chloride injection was intravenously given, once a day, until 7 days after flap pedicle surgery. The control group was not given PTX treatment. The value of blood perfusion (PU) was measured using laser-Doppler blood reperfusion image after flap transplantation, before and after pedicle division. RESULTS AND CONCLUSION: All 39 patients were completely cured and discharged, with no interruption experiments. Before pedicle division, PU value at distal flap in both drug and control groups were increased obviously, and the PU value in the drug group was significantly higher than control group (P < 0.05); after pedicle division, the PU value in the two groups were decreased, and there was no significant difference between the two groups (P > 0.05). Prior to pedicle division, the PU value of pedicle was gradually decreased and then increased in the drug group, and that in the control group was gradually increased. On the seventh day, the PU value of pedicle was stable in the drug group, and there was no significant difference between the two groups (P> 0.05); after pedicle division immediately, the PU values of pedicle were decreased in the two groups, and then the increase in the drug group was remarkable compared to control group (P < 0.05). Two sets of random flaps all survived, and skin defects were successfully repaired after pedicle division. PTX can markedly increase blood perfusion after random flap transplantation, promote flap survival and pedicle division in an early stage, and effectively shorten the healing time.

BACKGROUND: Random flap as a primary means of wound healing, is widely used at present, its blood circulation to establish the situation is also researched a lot, but not yet the system of random skin flap perfusion were observed and measured. In addition, the timing of pedicle division of a pedicle flap random is also a hot topic, but not yet a mature clinical testing method has been discovered to determine the best timing.OBJECTIVE: By means of laser Doppler blood perfusion imaging, this study was designed to dynamically observe random flap microcirculation, to understand the changes on random flap blood flow, and to determine the best timing of pedicle division. METHODS: A total of 18 cases were divided into traditional pedicle division group and early pedicle division group. Pedicle flap blood perfusion values were statistically measured immediately after surgery, at 3, 7,11,15, and 19 days after surgery, before division, immediately after division, and at 24 hours after pedicle division, 8-9 phases in total.RESULTS AND CONCLUSION: Distal blood perfusion value was increased with the time prolongation in both groups; while, the blood perfusion in various time phases was significantly different from that after surgery (P < 0.05); but, the blood perfusion was decreased immediately after surgery, which was still significantly compared with traditional pedicle division group (P< 0.05). There was no significant different in blood perfusion between early pedicle division and immediate after surgery of pedicle division (P > 0.05), but there was significant difference between 24 hours after pedicle division and immediate after surgery of pedicle division (P< 0.05). Blood perfusion values were less changed in both groups (P> 0.05). The ratio of both groups peaked before pedicle division and then gradually decreased after pedicle division. The best timing of pedicle division was the ratio of 1.2.

Objective To investigate the effect of cerebral ischemia on functional parameters of affected arteries and probe into the possible pathogenesis of ischemia-reperfusion injury.Methods Intraluminal suture ischemic model was used by occlusion of left middle cerebral artery in rats.Two hours later,the middle cerebral artery segments were isolated from both ischemia and control groups for measurement of changes in vessel diameter induced by increasing pressure and vasoactive compounds.And then,distensibility,myogenic tone,reactivity to 5-HT and ACh were calculated and compared between groups.Results In lower pressure range,ischemic vessels showed an increased myogenic tone(at 40 mm Hg,1 mm Hg=0.133 kPa,19.3%±0.4% vs 10.0%±0.2%,t=20.568,P=0.000)and decreased diameter.In higher pressure range,ischemic vessels showed an increased diameter.distensibility and decreased myogenic tone(at 120 mm Hg,12.0%±0.2% vs 21.8%±0.4%,t=-23.575,P=0.000).In normal pressure range,myogenic tone was not altered after ischemia. Both groups constricted to 5-HT and dilated to ACh,however,the response was significantly diminished after ischemla.Conclusion These findings demonstrate that contractile and diastolic function of affected artery was impaired after ischemia,a result that may contribute to ischemia-reperfusion injury by losing upstream cerebrovascular resistance and increasing perfusion on the microcirculation.

Objective:To investigate the safety and short-term efficacy of laparoscopic Nissen fundoplication in the treatment of refractory gastroesophageal reflux disease (rGERD).Methods:The clinical data of 61 patients underwent laparoscopic Nissen fundoplication from March 2018 to March 2022 in Jiangyin People′s Hospital were retrospectively analyzed. Among them, 14 patients had significant symptom relief after using proton pump inhibitor (PPI) before operation (group A), 30 patients had partial symptom relief after using PPI (group B), and 17 patients had persistent symptoms despite regular treatment with double-dose PPI for more than 8 weeks (group C). The surgical outcomes and recovery were compared among the three groups.Results:For the 61 patients, the surgical time was (117.46 ± 28.50) min, the intraoperative blood loss was 23.00 (8.00, 34.00) ml, and the postoperative hospital stay was 3.00 (2.00, 5.00) d. There were no statistically significant differences in surgical time, intraoperative blood loss, postoperative hospital stay, concurrent hiatal hernia repair and mesh placement among the three groups ( P>0.05). No short-term severe complications such as abdominal bleeding, abdominal infection and gastrointestinal perforation occurred in any group. There were no statistical differences in satisfaction score, subjective relief of overall postoperative symptoms, reflux symptoms, PPI usage, dysphagia, abdominal distention, diarrhea or constipation among the three groups ( P<0.05). No upper abdominal pain, recurrence and reoperation occurred in the three groups. Conclusions:Laparoscopic Nissen fundoplication has a definite therapeutic effect on rGERD, with significant anti reflux effects. There are no serious complications after surgery, and there are no recurrence or reoperation.

Objective To explore the expression of long non-coding ribonucleic acid(lncRNA)metastasis-associated lung adenocarcinoma transcript 1(MALAT1)and nuclear paraspeckle assembly transcript 1(NEAT1)in the hippocampus and HT22 cells of hypoxia pre-acclimated(HPC)mice and their relationship with neuroprotection.Methods(1)Thirty-six male Institute of Cancer Research(ICR)mice were randomly divided into three groups according to the random number table method of complete randomization:the control group,the hypoxia group and the hypoxia preconditioning group,with 12mice in each group.Mice in the control group were not exposed to hypoxia,mice in the hypoxia group were exposed to hypoxia once,and mice in the hypoxia preconditioning group were exposed to hypoxia four times.Immediately after the end of hypoxia treatment,all mice were decapitated and killed and hippocampal tissues were isolated and preserved in groups.(2)HT22 cells were cultured in medium containing 10％foetal bovine serum and 100 U/ml penicillin-streptomycin.When cell confluence was greater than 90％,they were transferred to 24-well plates for culture and then processed in 2 batches.6 pmol disordered small interfering RNA(siRNA),MALAT1 siRNA(siMALAT1),NEAT1 siRNA(siNEAT1),siMALAT1+siNEAT1 were transfected into the negative control group,siMALAT1 group,siNEAT1 group,and siMALAT1+siNEAT1 group of the first batch of HT22 cells one by one by transfection reagent,and the blank group did not have any treatment;then they were cultured under normal conditions(5％CO2 and 95％air)for 48 h.In the second batch of HT22 cells,6 pmol of disordered siRNA,disordered siRNA,siMALAT1,siMALAT1,siNEAT1 and siNEAT1 were transfected one by one correspondingly to the negative control group and the negative control+oxygen-glucose deprived/reoxygen(OGD/R)group,siMALAT1 group,siMALAT1+OGD/R,siNEAT1 group,siNEAT1+OGD/R group.48 h after transfection,HT22 cells of negative control group,siMALAT1 group and siNEAT1 group were cultured under normal conditions(5％CO2 and 95％air),and the cells of negative control+OGD/R group,siMALAT1+OGD/R group and siNEAT1+OGD/R group were treated with OGD/R.That is,under low oxygen conditions(1％O2+5％CO2+94％N2)exposure for 8 h,and then culture under normal conditions for 16 h.(3)The real-time fluorescence quantitative polymerase chain reaction(PCR)and Western blot was used to determine the expression of MALAT1,NEAT1,N-methyl-D-aspartate receptor subunit 2B(NR2B)messenger RNA(mRNA)and NR2B protein in the hippocampus of mice,the relative expression levels of NR2B mRNA and NR2B protein after transfection of HT22 cells in each group,and the relative expression levels of haemoglobin breakdown products and activated cysteine protease protein 3 after transfection and OGD/R of HT22 cells in each group.The survival rate of HT22 cells in each group was calculated.Results(1)The differences in relative expression of MALAT1(F=43.92),NEAT1(F=506.4),NR2B mRNA(F=50.64)and NR2B protein(F=41.24)in the hippocampus of mice in the three groups were statistically significant(all P＜0.05).The relative expression of MALAT1([1.68±0.06]vs.[1.00±0.08]),NR2B mRNA([1.26±0.06]vs.[1.00±0.01]),and NR2B protein([1.47±0.05]vs.[1.00±0.01])was increased in the hypoxia group as compared to the control group(all P＜0.05),whereas the relative expression of NEAT1([1.02±0.10]vs.[1.00±0.03])were not statistically significant(P＞0.05),and the relative expression of MALAT1([1.12±0.13]vs.[1.00±0.08])and NEAT1([2.88±0.10]vs.[1.00±0.03])were increased in hypoxic preconditioned group.Compared with hypoxia group,the relative expression of NR2B mRNA([0.54±0.07]vs.[1.26±0.06])and NR2B protein([1.17±0.07]vs.[1.47±0.05])were decreased(both P＜0.05).(2)The differences in the relative expression of NR2B mRNA(F=36.92)and NR2B protein(F=56.98)after transfection of HT22 cells in the five groups were statistically significant(both P＜0.05).Compared with the negative control group,siMALAT1 group(NR2B mRNA:[2.04±0.08]vs.[0.94±0.04],NR2B protein:[1.72±0.13]vs.[0.93±0.02]),siNEAT1 group(NR2B mRNA:[2.15±0.13]vs.[0.94±0.04],NR2B protein:[1.87±0.46]vs.[0.93±0.02]),siMALAT1+siNEAT1 group(NR2BmRNA:[2.09±0.16]vs.[0.94±0.04],NR2B protein:[2.07±0.30]vs.[0.93±0.02])showed the relative NR2B mRNA and NR2B protein expression were increased(all P＜0.05).(3)Differences in relative expression of haematopoietin breakdown product(145/150 kDa)protein(F=12.43),haematopoietin breakdown product(120 kDa)protein(F=7.15),and activated cysteamine protease protein 3 protein(F=6.61)were statistically significant in the 6 groups of HT22 cells transfected and treated with OGD/R(all P＜0.05).Compared with the siMALAT1 group,the siMALAT1+OGD/R group had 145/150kDa([1.42±0.48]vs.[0.85±0.34]),120 kDa([1.33±0.37]vs.[0.52±0.19])haematopoietin catabolism products and activated cysteamine protease protein 3([2.43±0.35]vs.[1.15±0.24])relative expression increased(all P＜0.05);compared with the negative control+OGD/R group,the siMALAT1+OGD/R group showed an increase in 145/150kDa([1.42±0.48]vs.[1.23±0.17]),120 kDa([1.33±0.37]vs.[0.80±0.21])relative expression of haematopoietin breakdown products and activated cysteamine protease protein 3([2.43±0.35]vs.[1.46±0.39])increased(all P＜0.05);compared with the siNEAT1 group,the siNEAT1+OGD/R group had a higher expression of 145/150 kDa([1.28±0.44]vs.[0.87±0.32]),120 kDa([0.81±0.36]vs.[0.63±0.16])relative expression of haematopoietic proteolytic products and activated cysteamine protease protein 3([1.51±0.45]vs.[1.01±0.27])increased(all P＜0.05).(4)The difference in HT22 cell survival rate among the 6 groups was statistically significant(F=5.54,P＜0.05).Compared with the negative control group,HT22 cell survival was decreased in the siMALAT1,siNEAT1,siMALAT1+OGD/R and siNEAT1+OGD/R groups([0.65±0.40],[0.76±0.35],[0.24±0.17],[0.23±0.16]vs.[0.84±0.04],all P＜0.05);cell viability was reduced in the siMALAT1+OGD/R group compared with the siMALAT1 group([0.24±0.17]vs.[0.65±0.40],P＜0.05);and cell viability was reduced in the siNEAT1+OGD/R group compared with the siNEAT1 group([0.23±0.16]vs.[0.76±0.35],P＜0.05).Conclusion HPC increased the expression of MALAT1 and NEAT1 in the hippocampus of mice,and MALAT1 and NEAT1 may participate in the neuroprotective effect of mice after ischemia and hypoxia by affecting the expression of NR2B.

OBJECTIVETo investigate the value of serum IgA/C3 ratio in the diagnosis of IgA nephropathy and explore its relationship with the clinicopathological features of the patients.

METHODSSixty-six patients with IgA nephropathy, 111 with other glomerular diseases, and 40 healthy control subjects without kidney disease were tested for serum IgA and C3 levels using CRM470 adjusted standardized immune turbidimetric method, and the IgA/C3 ratio was calculated. According to Oxford and Lee's classification criteria, we analyzed the pathological grades of the renal biopsy samples from patients with IgA nephropathy. The ROC curve was used to assess the value of serum IgA and IgA/C3 ratio in predicting IgA nephropathy.

RESULTSPatients with IgA nephropathy had an elevated serum IgA/C3 ratio than those with other glomerular diseases and the control subjects, with an area under the ROC curve of 0.776. An elevated serum IgA/C3 ratio was not found to significantly correlate with the pathological grade of renal biopsy samples in patients with IgA nephropathy.

CONCLUSIONIn the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.

Biopsy ; Case-Control Studies ; Complement C3 ; analysis ; Glomerulonephritis, IGA ; blood ; diagnosis ; Humans ; Immunoglobulin A ; blood ; Kidney ; pathology