OBJECTIVE:To observe the clinical effects of Sanqi shengji ointment (SQSJO) preventing skin flap necrosis. METHODS:75 cases of local flap transfer in head and facial operation were randomly divided into blank group,treatment group, control group and with 25 cases in each group. After surgery,except for conventional treatment,treatment group was coated with SQSJO additionally;control group was additionally treated with rh-bFGF. The dressing was changed with iodophor in blank group conventionally till the wound healed. The skin flap necrosis,healing course,local skin temperature,swelling disappearance time and wound healing time were observed and compared among 3 groups after surgery. RESULTS:Compared with blank group,the incidence of skin flap necrosis decreased significantly in other 2 groups,there was statistical significance(P<0.05). There was sta-tistically significant difference in healing rate ameng 3 grups(P<0.05);there was no statistical significance in local skin tempera-ture among 3 groups after surgery (P>0.05);the flap swelling disappearance time and wound healing time of treatment group were significantly shorter than those of control group and blank group;there was statistical significance (P<0.05). CONCLU-SIONS:SQSJO can prevent skin flap necrosis accurately after flap transfer,and has no significant difference from rh-bFGF in pre-vention effect. Moreover,it is better than rh-bFGF in improving local swelling and shortening wound healing.

Object To study the inhibitory effect of Anti-EB Virus Liquor (AEVL) on EB (Epstein-Barr) virus antigen expression and its cytotoxicity. Methods The effect of AEVL on Raji cell early antigen (EA) expression and B 95-8 cell virus capsid antigen (VCA) expression was assayed by indirect fluorescent technique; the cytotoxicity on nasopharyngeal carcinoma CNE 2 cells was determined by MTT method. Results At the non-toxic concentration, AEVL had markedly inhibitory effect on Raji cell EB-virus, IC 50 was 0.667 mg/mL and showed powerfully inhibitory effect on B 95-8 cell EB-virus VCA expression, IC 50 was 0.89 mg/mL; and it had strongly inhibitory effect on B 95-8 cell EB virus VCA expression stimulated by sodium n-butyric acid, IC 50 was 1.4 mg/mL. The IC 50 of AEVL on human nasopharyngeal carcinoma CNE 2 cell was 7.57 mg/mL. Conclusion AEVL could inhibit EB virus antigen expression and have cytotoxicity on nasopharyngel carcinoma cells at high concentration.

Objective To investigate the growth inhibition of human osteosareoma cell line(MG-63) intervened by Hydralazine and 5'-aza-2'-deoxycytidine (5-Aza-CdR),and the effect on the mRNA expression of gene WW domain-containing oxidoreductase (WWOX).Methods Certain volume of 5 × 104/ml of human osteosarcoma cell line MG-63 in logarithmic growth phase were added into 96-well plate.There were Hydralazine group (drug concentration,0.1,1.0,10 μmol/L),5-Aza-CdR group (drug concentration,5,10,20 μmol/L),Hydralazine combined with 5-Aza-CdR group (drug concentration,0.1 μmo/L + 5 μmol/L,1.0 μmol/L + 10 μmol/L,10 μmol/L + 20 μmol/L) and control group (culture medium).Methyl thiazol tetrazolium(MTT) colorimetric methods were used to test the growth inhibition of MG-63 cells intervened by different concentrations of Hydralazine and 5'-aza -2'-deoxycytidine (5-Aza-CdR).Flow cytometry AnnexinV-FITC/PI methods were used to assay the effects of Hydralazine and 5-Aza-CdR inducing apoptosis in osteosarcoma cells in vitro.Real-time polymerase chain reaction (Real-Time PCR)methods were used to detect amplification of WWOX mRNA induced by Hydralazine combined with 5-Aza-CdR or alone.Western-blotting methods were used to examine the expression of WWOX in MG-63 cells.Results Hydralazine and 5-Aza-CdR effectively inhibited the growth of MG-63 cells in a concentration and time-dependent manner.Combined effect was more obvious.Further more the expression levels of WWOX mRNA and protein were increased significantly in combined groups as compared with other groups.Conclusion Hydralazine and 5-Aza-CdR could effectively inhibit the proliferation of MG-63 cells and induce apoptosis which is concurrent with the promotion of the expression of WWOX.The mechanism may be that Hydralazine/5-Aza-CdR effectively cause the demethylated of WWOX gene CpG-rich promoter regions,leading to the high expression of WWOX and inhibit the growth of MG-63 cells.The use of hydralazine in the treatment of osteosarcoma is worthy of further investigation.

Aim To investigate apoptosis induced by 3,3'-diethyl-9-methylthia-carbocyanine iodide(DMTCCI) , an inhibitor of DNA primase found in our previous study, and the mechanism of DMTCCI in human myelogenous leukemia HL-60 cells. Methods HL-60 cells were cultured in RPMI-1640 medium and treated with different concentrations of DMTCCI. MTT assay was used to detect growth inhibition.Flow cytometry and DNA ladders were used to detect apoptosis. Western blotting was used to observe the expression of survivin, Bcl-xL, Bad, Bax, Bcl-2, caspase-9, caspase-3, caspase-6, PARP, DFF45 and lamin B protein. Caspase-3 activity was measured by ApoAlert Caspase-3 Assay Kit. Results DMTCCI inhibited proliferation of human leukemia HL-60 cells with IC50 value of 0. 24 μmol · L-1. The results of flow cytometry and DNA ladders showed that DMTCCI could induce apoptosis of HL-60 cells. The expression levels of protein survivin and Bcl-xL were down-regulated, Bad and Bax were up-regulated,while Bcl-2 protein had no change in response to DMTCCI treatment in HL-60 cells. Treatment of HL-60cells with DMTCCI induced the proteolytic cleavage of caspase-9, caspase-3, caspase-6, PARP, DFF45and lamin B protein. Caspase-3 activity apparently increased at 3 h and reached a peak at 12 h after exposure to 1 μmol · L-1 of DMTCCI in HL-60 cells. Conclusion DMTCCI inhibited proliferation and induced apoptosis of human leukemia HL-60 cells. Bcl-2 family proteins, survivin and caspases family proteins might playa role in the apoptosis process induced by DMTCCI.

Objective To investigate the effects of different concentrations of botulinum toxin type A on hypertrophic scar fi-broblasts ,and to explore the molecular mechanism of botulinum toxin type A in the treatment of scar and prevention of postopera-tive scar hyperplasia .Methods Different concentrations of botulinum toxin A (0 .01 ,0 .1 ,1 U/L and 10 U/L) were used on hyper-trophic scar fibroblasts for 24 hours ,to observe the changes of cell adhesion and cytoskeleton under laser confocal microscopy .MTT and flow cytometry were used to detect the proliferation ,apoptosis and cycle of change ,at the same time real time fluorescence quantitative PCR and Western blot were conducted to detected the expression of TGF-β,matrix metalloproteinase MMP-1 ,MMP-2 and MMP-9 gene and protein expression changes .Results With the increase of botulinum toxin A dose ,the number of cell adhesion and cytoskeletal fluorescence intensity decreased ,cell proliferation ability decreased and mainly blocked at G0-G1 phase ,and the ap-optosis also increased with the dose increased .The results of qPCR and Western blot showed that MMP-1 and MMP-9 gene and protein were highly expressed with the increase of botulinum toxin A dose ,while TGF-βand MMP-9 showed low expression .Con-clusion Botulinum toxin A can inhibit the proliferation of hypertrophic scar fibroblasts and inhibit the expression of MMP-1 and MMP-2 ,which can inhibit scar formation .It plays a positive role in the treatment of scar .

Objective:To investigate the predictive value of procalcitonin to platelet ratio (PPR) and C-reactive protein to albumin ratio (CAR) in severe acute pancreatitis (SAP) and the value of SAP and concomitant acute liver injury (ALI).Methods:Total of 195 patients with AP from June 2021 to December 2022 from 374 patients were screened for inclusion in the study and were divided into non-severe acute pancreatitis (NSAP) and SAP groups. The ALI group was divided into non-acute liver injury (NALI) and ALI groups according to ALI criteria, and then into hepatocellular ALI subgroup, cholangiocellular ALI subgroup and mixed ALI subgroup. Laboratory tests for procalcitonin (PCT), C-reactive protein (CRP), albumin and platelet (PLT) were completed within 48 h. Risk factors for SAP, ALI and each subgroup of ALI were analysed by binary logistic regression. Subject work characteristic (ROC) curves were plotted and the optimal thresholds for PPR and CAR were calculated. The predictive value of PPR, CAR and their combination for SAP, ALI and each type of ALI was determined.Results:The AUCs for predicting SAP by plotting ROC curves and calculating the bedside index score of acute pancreatitis severity (BISAP score), PPR, CAR, PPR combined with CAR, PPR combined with BISAP score, CAR combined with BISAP score and combined PPR, CAR and BISAP score were 0.82, 0.85, 0.79 and 0.86. The areas under the ROC curves for PPR, CAR and combined prediction of ALI were 0.81, 0.85 and 0.88, respectively; the areas under the ROC curves for PPR, CAR and combined prediction of hepatocellular ALI were 0.93, 0.77 and 0.92, respectively; and the areas under the ROC curves for PPR, CAR and combined prediction of cholangiocellular ALI were 0.76, 0.76 and 0.77, respectively. The area under the ROC curves for PPR, CAR and combined prediction of mixed ALI were 0.83, 0.76 and 0.82Conclusions:Elevated PPR and CAR are risk factors for SAP and for the development of ALI in AP. PPR has better predictive value than CAR for hepatocellular and mixed ALI, and CAR has better predictive value than PPR for cholangiocellular ALI.

OBJECTIVETo study whether recombinant adeno-associated virus (rAAV) mediated foreign gene, LacZ, could pass the blood brain barrier by intra-carotid artery delivery and express in vivo in ischemic brain of the focal embolic stroke rats to investigate a possibility of delivering foreign gene through carotid artery to treat acute ischemic stroke.

METHODSThe carotid artery territory in 41 rats was embolized with or without arterial-like fibrin rich clots to make a model of focal embolic stroke rat. rAAV containing LacZ gene (rAAV-LacZ) was constructed in 293 cells by calcium phosphate cotransfection. The rats were assigned to one of the following treatments: 1 control (without embolism) groups, including PBS treated (n = 6), pLacZ treated (n = 6 ) and rAAV-LacZ treated (n = 6): 2 embolic groups, including embolism + PBS (n =7),embolism + pLacZ (n = 8) and embolism + rAAV-LacZ (n = 8). Brains were cryosectioned and kappa-Gal stain was performed at 2, 4, and 8 weeks, respectively, after transfection, and then infarct volume was measured and the percentage of LacZ staining-positive cells was calculated.

RESULTSIn all the control groups and embolism + PBS treated animal, no kappa-Gal staining-positive cells were found, but in embolism + pLacZ (n = 8) and embolism+rAAV-LacZ groups a lot of kappa-Gal staining-positive cells were found. The expression cells were in the tissues around the infarction. The gene expression persisted only nearly four weeks in embolic group with pLacZ. In the embolic group with rAAV-LacZ the expression was very stable during the experiment course (eight weeks) and the percentage of the expressed cells was significantly higher than that of its contralateral areas at the same time points, respectively.

CONCLUSIONSThe plasmid vector and rAAV could enter the brain through the ischemia-damaged blood barrier and foreign gene can be expressed in brain. The positive gene expression is mainly in the peripheral areas of the infarction. rAAV as a permanent expression vector may ultimately be used for gene therapy of human ischemia cerebravascular diseases.

Animals ; Blood-Brain Barrier ; Brain ; virology ; Carotid Arteries ; Dependovirus ; genetics ; Genetic Therapy ; Genetic Vectors ; Intracranial Embolism ; therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Stroke ; therapy

Objective:To investigate the protective effect of overexpressed tripartite motif containing (TRIM27) on severe acute pancreatitis (SAP) in mice and its possible mechanism.Methods:Twenty-four mice were randomly divided into the sham operation + control virus group (AAV-GFP group), sham operation + overexpression of TRIM27 group (AAV-TRIM27 group), SAP + control virus group (SAP+AAV-GFP group), SAP + overexpression of TRIM27 group (SAP + AAV-TRIM27 group), with 6 mice in each group. SAP model of mice was established by intraperitoneal injection of L-arginine (4 mg/kg). The sham operation group was injected with equal volume of normal saline, and the virus group was injected with control or TRIM27 overexpression adeno-associated virus (2×10 11 μg/ per mice). The serum and pancreatic tissue samples were collected 72 h after modeling. The levels of serum amylase, lipase, tumor necrosis factor α (TNF-α), interleukin-1b (IL-1b), IL-6, macrophage chemoattractant protein-1 (MCP-1) and the expression of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) in pancreatic tissue were detected by enzyme-linked immunosorbent assay. Hematoxylin eosin staining was used to observe the pathological damage of pancreatic tissue. The expressions of myeloperoxidase (MPO) and Ly6g positive inflammatory cells in mouse pancreas were observed by immunohistochemistry. The expression of p-p65, p65, p-ASK1, ASK1, p-JNK, JNK, p-p38 and p38 in pancreatic tissue were detected by Western blot. Results:The expression of TRIM27 in pancreatic of mice was significantly down regulated after SAP ( P<0.05); after overexpression of TRIM27 by adeno-associated virus, the expression of TRIM27 in mouse pancreas was significantly up-regulated ( P<0.05). There was no significant difference in the indexes of mice between the AAV-GFP group and AAV-TRIM27 group ( P>0.05). Compared with the SAP + AAV-GFP group, the levels of serum amylase, lipase, TNF-α, IL-1b, IL-6 and MCP-1 in mice of the SAP + AAV-TRIM27 group were significantly decreased, MDA in pancreatic tissue was decreased, SOD and GSH were increased, MPO and Ly6g inflammatory cells were significantly decreased, and p-p65, p-ASK1, p-JNK, and p-p38 protein expression were down regulated. Conclusions:Overexpression of TRIM27 alleviates SAP in mice by inhibiting inflammatory response and oxidative stress, and its mechanism may be through inhibiting NFκB/MAPK signaling pathway.

Objective:To investigate the predictive value of dynamic platelet and hemagglutination-related parameters in patients with acute pancreatitis (AP).Methods:The patients admitted to the Department of Emergency Surgery in the First Affiliated Hospital of Zhengzhou University from January 2020 to December 2020 were analyzed. According to the inclusion criteria and exclusion criteria, patients with AP were retrospectively enrolled. According to the Chinese Guidelines for the Diagnosis and Treatment of Acute Pancreatitis (Shenyang, 2019), the patients were divided into two groups: severe acute pancreatitis (SAP group) and non-severe acute pancreatitis (non-SAP group) [including mild acute pancreatitis (MAP) and moderate severe acute pancreatitis (MSAP)]. A normal distribution of the maximum and mean aggregation rates of dynamic platelets (arachiidonic acid), plateletcrit (PCT) and bedside index for severity in acute pancreatitis (BISAP) scores and other measurement data were tested by t test, while measurement data of prothrombin time (PT), fibrinogen (FIB) and D-dimer that did not conform to normal distribution were tested by Mann-Whitney U test. χ 2 test was used for the counting data such as sex, age and etiology of patients in the two groups. The prognostic value of statistically significant indicators for non-SAP group and SAP group was further analyzed by receiver operating characteristic (ROC) curve. Results:A total of 146 patients with AP were enrolled, including 50 patients in SAP group and 96 in non-SAP group. The maximum and average aggregation rates of dynamic platelet (aracidonic acid) in the SAP group were (71.76±17.62) % and (67.91±18.10) %, PT (12.02±1.33) s, FIB (4.76±2.08) g/L, D-dimer (3.75±6.04) μg/L, PCT (0.23±0.08) %, and BISAP scores (1.42±1.18), which were all significantly higher than those in the non-SAP group [the maximum and average aggregation rates of dynamic platelet (arachiidonic acid) (46.65±20.11) % and (42.50±20.71) %, PT (11.50±1.51) s and FIB (3.91±1.48) g/L, D-dimer (1.00±1.37) μg/L, PCT (0.19±0.06) %, BISAP scores (0.45±0.66)] (all P<0.05). According to area under the ROC curve, the maximum and average aggregation rates of dynamic platelets (arachiidonic acid) in serum of patients with SAP were 0.83 and 0.82, respectively, and the sensitivities were 0.56 and 0.68, respectively. The specificity was 0.99 and 0.81, respectively, which was better than PT, FIB, D-dimer, PCT and BISAP scores in predicting the severity of AP. Conclusions:The maximum and average aggregation rates of dynamic platelets (arachidonic acid), PT, FIB, D-dimer, PCT and BISAP scores can be used as predictors of the severity of acute pancreatitis. The maximum and average aggregation rates of dynamic platelets (arachiidonic acid) were the best in predicting the severity of AP.