Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1α) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1α anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1α sense oligonueleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5 % glucose solution caudalis veins injection, group C and group D rats received HIF-1α ASODN and HIF-1α SODN caudalis veins injection, respectively(0.25 mg/kg). The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD 18 positive neutrophil cell was (44.93±3. 60)% in group B, (18.66±1.52)% in group A, (31.66±4.72)% in group C,(51.00±5.66)% in group D. Compared with each other groups,the differences are statistically significant (F= 42. 46, P<0.001). The number of positive staining cells of retinal leukocyte was (46.16±10.68)in group A, (133.83±20.43)in group B, (99.83±9.28)in group C, (121.33±10. 23) in group C. Compared group B with group C,the number of positive staining cells raised about 2.89 times;compared group B with group C and D,the differences are statistically significant (P= 0.12, 95% confidence interval -3.69～28. 69 ). Conclusions In vivo, HIF-1α can decreased the expression of CD18 on neutrophils from diabetic rats' peripheral blood and the collection of retinal leukos- tasis in the diabetic animals. HIF-1α may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy.

Objective To explore the effects of ferulic acid on gastric cancer cell line MGC-803 proliferation;To discuss the mechanism of apoptosis induced by ferulic acid. Methods Ferulic acid with a variety of concentrations was used to treat gastric cancer cell line MGC-803 for 24, 48, 72 h;MTT experiment was used to detect the growth of gastric cancer cells. After gastric cancer cell line MGC-803 were treated with ferulic acid with a variety of concentrations (0, 5, 7.5, 10 mg/mL) for 48 h, all gastric cancer cells were collected and stained by Annexin V-FITC/PI for the detection of apoptosis by flow cytometry. RT-qPCR and Western-blot methods were used to detect mRNA and protein levels of Caspase-3, Caspase-9, Bax, Bcl-2 and Xiap. Results Compared with the control group, the OD value of cell line MGC-803 treated by ferulic acid with a variety of concentrations (5, 7.5, 10, 12.5 mg/mL) decreased significantly;the anti-tumor effect of ferulic acid was obvious;IC50 of 24, 48, and 72 hours after treated by ferulic acid was 12.93, 9.73 and 5.52 mg/mL respectively. Cell lineMGC-803 treated by ferulic acid with a variety of concentrations (5, 7.5, 10 mg/mL) after 48 h could induce the apoptosis of MGC-803 cells, up-regulate mRNA and protein levels of Caspase-3, Caspase-9, and Bax, down-regulate mRNA and protein levels of Bcl-2 and Xiap. Conclusion Ferulic acid can inhibit the proliferation of MGC-803 cells effectively and induce the apoptosis of MGC-803 cells, which mechanism is related to mitochondria apoptosis pathway and the down-regulation of Xiap expression.

Objective To explore the mechanisms and the significance of compatibility of Heixiaoyao Powder by observing the effect of Heixiaoyao Powder on mice model of senile dementia induced by scopolamine. Methods Kunming mice were randomly divided into five groups including control group, model group, Heixiaoyao Powder group, Xiaoyao powder group and streamlined formula group, with 10 mice of each group. Scopolamine hydrobromide intraperitoneal injection was adopted to make memory impairment in mice model, and followed by drugs treatment with corresponding decoction of the crude drug 2.314 g /(kg·d). Control group and model group were given normal saline orally once a day, all groups were treated for 10 days continuously. After 6 and 8 days of the administration, water maze test, stair jumping test and darkness avoidance test were conducted to test behavior proficiency, and the activity of acetylcholinesterase (AchE) activity and choline acetyltransferase (ChAT) in brain tissue were tested also. Results Compared with model group, Heixiaoyao Powder group, Xiaoyao Powder group and streamlined formula group could significantly reduce the error number and shorten latency in the water maze test, decrease the time of electrical shocks and prolong the incubation period of time in both stair jumping test and darkness avoidance test, and could increase ChAT activity and lower AchE activity in brain tissue of mice (P<0.01 or P<0.05), especially the effects of Heixiaoyao Powder group. Conclusions Heixiaoyao Powder and its analogous formula have the function of preventing and treating senile dementia.

Alzheimer's disease （AD） is a neurological disease highly related to age， which is the main cause of senile dementia and the most common disease leading to the loss of daily living ability of the elderly. AD brings heavy mental burden and economic pressure to patients， families， and society. Traditional Chinese medicine （TCM） ascribes AD to category of "dementia"， believing that the treatment should start from kidney because kidney deficiency is the root cause. Combined with the physiological and pathological characteristics of liver， this paper proposed that liver-kidney homology was an important idea for the prevention and treatment of AD. The main pathological manifestations of AD were amyloid β-protein （Aβ） deposition and neurofibrillary tangles （NFT）， and the pathogenesis was complex. A growing number of studies showed that immune inflammation played an important role in the pathogenesis of AD. The important target of treating AD was the regulation of neuro-immune inflammation through the nuclear factor kappa B （NF-κB）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/Caspase-1/interleukin-1β （IL-1β） signaling pathway. Based on the idea of liver-kidney homology， this paper selected the representative formula Hei Xiaoyaosan to explore its effect on the prevention and treatment of AD and the mechanism from the perspective of regulating NF-κB/NLRP3/Caspase-1/IL-1β signaling pathway and inhibiting neuro-immune inflammation， expecting to further promote the in-depth study on the prevention and treatment of AD， and provide references for the prevention and treatment of AD by TCM.

Objective: To explore the effects of scar excision combined with negative-pressure on repair of hypertrophic scar in burn children. Methods: From October 2010 to August 2016, 25 children with hypertrophic scar after deep burn were hospitalized, with scar course ranging from 3 months to 11 years and scar area ranging from 35 to 427 [83(51, 98)]cm². A total of 35 scars of 25 children were located in trunk (11 scars), upper limb (11 scars), and lower limb (13 scars). All children received scar excision operation and negative-pressure treatment (negative-pressure value ranged from -40 to -20 kPa), among which 6 cases received scar excision operation and negative-pressure treatment for two times for further removal of scars. After scar excision, electronic spring scale was used to measure the tension of the incision. The tension value of children ranged from 3.43 to 23.84 [7.16 (5.59, 9.12)] N, and then the incision was closed with appropriate suture according to the value of the tension. The incision with smaller tension was firstly opened on post operation day (POD) 8. After removing the suture, negative-pressure was conducted to POD 14. The incision with larger tension was firstly opened on POD 12. After removing the suture, biological semi-membrane was used to reduce tension to POD 16. All healed incisions were performed with anti-scar treatment for 1 year and relaxation and fixation for 3 months. General condition of the incision was observed after operation. The reduction percentage of scar area was calculated half-year after operation. The Patient and Observer Scar Assessment Scale was used to record the overall score of scar and scar score of trunk, upper limb, and lower limb before operation and half-year after operation. Data were processed with paired t test and Wilcoxon rank sum test. Results: After removing the suture, all incisions of children healed well without redness, effusion, and rupture. Half-year after operation, the appearance and deformity of incision were obviously improved, and the symptoms including pruritus and pain were basically relieved. Half-year after operation, the scar area of children ranged from 0 to 174 [21(9, 47)]cm^2, which was significantly decreased as compared with that before operation (Z=-5.16, P<0.05). The reduction percentage of scar area ranged from 36% to 100% [(73±19)%]. Half-year after operation, the overall score of scar and scar score of trunk, upper limb, and lower limb of children were obviously decreased as compared with those before operation (with t values from 6.42 to 17.37, P values below 0.05). Conclusions Scar excision combined with negative-pressure treatment has a good clinical effect on repair of hypertrophic scar in burn children, which is suitable for clinical application.

Objective: To establish a method for repairing extremities with extensively deep burn using large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin and observe its effect. Methods: Medical records of two male patients with extremely extensive deep burn admitted to our hospital from May to November in 2018 were retrospectively analyzed. Two patients aged 44 and 25 years respectively, with total burn area of 90% and 97% total body surface area (TBSA) and full-thickness burn area of 85% and 70% TBSA, respectively. Preoperatively, the surgical area on the extremities was calculated to estimate the necessary amount of allogeneic scalp and Meek miniature skin. The large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin was prepared according to the methods described as follows. Thin medium-thickness fresh scalps with 3% TBSA and 0.30-0.35 mm in depth were harvested from each donor and spliced into a large piece with epidermis upward by spraying Meek glue. Then the spliced scalp was punched after covered with a single-layer gauze. Autologous microskin was transported onto the dermis of fresh large piece of allogeneic scalp by traditional floating method. Bilateral extremities with full-thickness burn of two patients were selected for self-control. The left upper extremity was denoted as treatment group while the right upper extremity was denoted as control group in Patient 1. The right lower extremity was denoted as treatment group while the left lower extremity was denoted as control group in Patient 2. Wounds in the treatment group were treated with fresh large piece of allogeneic scalp spliced by Meek glue and autologous microskin with expansion ratio of 1∶15 after escharectomy, while wounds in control group received grafting of Meek miniature skin with expansion ratio of 1∶6 and or 1∶9 after escharectomy. The donors of allogeneic scalp were 32 males who were the relatives or friends of the patients, aged 21-50 years, with scalp area of (548±48) cm². The healing conditions of donor sites of scalp were observed on post operation day 10, and were followed up within 3 months after operation to observe whether forming alopecia and hypertrophic scar or not. Wound healing condition was evaluated during follow-up in post operation week (POW) 2-5 and 4 months after operation. Wound coverage rates were calculated in both treatment and control groups in POW 2, 3, 4, and 5. Results: The donor sites of all allogeneic scalp of donors healed completely on post operation day 10. There was no alopecia or hypertrophic scar within 3 months after operation for follow-up. In POW 2, allogeneic scalp grafts basically survived in treatment group without obvious exudation, and most of the Meek miniature skin survived in control group with obvious exudation. Part of allogeneic scalp grafts dissolved and detached in treatment group in POW 3, and the surviving grafts scabbed. The eschar detached and new epithelium was observed in treatment group in POW 4 and 5. In POW 3-5, surviving Meek miniature skin in control group creeped and was incorporated, and the wounds shrank. Hypertrophic scar was observed in both treatment and control groups 4 months after operation, without obvious difference in scar as a whole. The wound coverage rates were respectively 84%-98% and 76%-92% in treatment group of two patients in POW 2-5, close to or higher than those of control group (35%-97% and 28%-81%, respectively). Conclusions The study establishes a novel method for splicing fresh allogeneic scalps into a large piece as the covering of microskin, which has good effect for repairing extensively deep burn wounds. Considering that allogeneic skin is scarce, this method may be a new option in clinical treatment for extensively deep burn patients.

Objective To explore the delayed discount and personality characteristics of impulsive and premeditated male prisoners and their correlation.Methods A total of 120 male prisoners with aggressive behaviors were tested by Self-made General Situation Questionnaire,Impulsive / Premeditated Aggression Scale (IPAS),NEO Five-Factor Inventory (NEO-FFI),Delay Discount Task (DDT).The effective participants were ninety,including 51 in impulsive group and 39 in premeditated group.Analyzed the difference of delayed discount and personality characteristics between the two group.Results (1) In the delayed discount task,the delay discount rat (K) of the impulsive violence group (0.0278 ± 0.0555) was significantly higher than that of the premeditated violence group (0.0042±0.0078).The difference was statistically significant (t=3.004,P =0.004).(2) The scores of agreeableness (35.84 ± 4.08) and conscientiousness (37.04±4.83) in impulsive violence group were lower than that in premeditated violence group(38.46±5.53,42.31±5.96),and the difference was statistically significant (t=-2.587,P=0.011;t =-4.634,P＜0.01).(3) Correlation analysis showed that K value was not significantly correlated with all dimensions of the big five personality(P＞0.05).Conclusion Male impulsive prisoners have faster tendency to delay discounting compared with male premeditated prisoners.Lower agreeableness and conscientiousness are the personality characteristics of male impulsive prisoners.

ObjectiveTo investigate the effect of Danzhi Xiaoyaosan on the phosphorylation of tau protein and different sites of glycogen synthase kinase-3β （GSK-3β） and phosphoseryl/suanyl phosphate protein phosphatase 2A （PP2A） in the hippocampus of rats with Alzheimer's disease （AD） and its mechanism. MethodThe rat model of AD was established by injecting okadaic acid into the bilateral hippocampus of 90 male Wistar rats in SPF grades. The rats with successful modeling were selected and randomly divided into model group， aricept group （0.5 mg·kg^-1）， and Danzhi Xiaoyaosan high， medium， and low groups （17.55， 8.77， and 4.38 g·kg^-1）， and then gavaged for 42 d， once a day. Morris water maze was used to detect the learning and memory ability of rats， Nissl's staining was used to observe the morphological structure of neurons in the hippocampus， and Real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of tau protein， GSK-3β， and PP2A. Western blot was used to determine the protein expression levels of tau protein， GSK-3β， and PP2A. ResultAs compared with the control group， the learning and memory abilities of the rats in the model group were significantly decreased （P<0.01）， and the hippocampal CA3 region cells had abnormal structure， disorderly arrangement， and decreased number. The expression levels of GSK-3β mRNA， GSK-3β， p-GSK-3β-Tyr216， p-PP2A， and p-tau were increased in the model group as compared with the control group （P<0.01）， and those of p-GSK-3β-Ser9 and PP2A decreased significantly （P<0.01）. As compared with the model group， the learning and memory ability of the Aricept group and the Danzhi Xiaoyaosan groups were improved （P<0.05， P<0.01）， and the cell morphology and the number of hippocampal CA3 regions were better. The mRNA expression levels of PP2A and tau in the Aricept group were significantly up-regulated （P<0.05）， the mRNA expression level of GSK-3β was significantly down-regulated （P<0.01）， and the protein expression levels of GSK-3β， p-GSK-3β-Tyr216， and p-PP2A were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of PP2A in the high-dose Danzhi Xiaoyaosan group was significantly up-regulated （P<0.01）， and that of GSK-3β was significantly down-regulated （P<0.01）， whereas the protein expression levels of p-PP2A， p-GSK-3β-Tyr216， and p-tau were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of GSK-3β was significantly down-regulated in the medium-dose Danzhi Xiaoyaosan group （P<0.01）， the protein expression levels of GSK-3β， p-GSK-3β-Tyr216， and p-tau were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of PP2A was significantly up-regulated in the low-dose Danzhi Xiaoyaosan group （P<0.01）， and that of GSK-3β was significantly down-regulated （P<0.01）， whereas the protein expression levels of GSK-3β and p-GSK-3β-Tyr216 were down-regulated （P<0.05， P<0.01）， and those of p-GSK-3β-Ser9 and PP2A were significantly up-regulated （P<0.01）. ConclusionDanzhi Xiaoyaosan can improve the learning and memory ability of rats with AD， and its mechanism may be related to the regulation of the activities of GSK-3β and PP2A protein-related sites and the phosphorylation of tau protein.

ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in regulating the tumor necrosis factor receptor superfamily member 6 （Fas）/Fas ligand （FasL）/cysteine protease-8 （Caspase-8）/cysteine protease-3 （Caspase-3） signaling pathway to intervene in neuronal apoptosis and prevent Alzheimer's disease （AD）. MethodNinety SPF-grade SD male rats of 4 months old were selected and randomly grouped as follows： 10 rats in the blank group， 10 rats in the sham group （bilateral hippocampus injected with 1 μL normal saline）， and 70 rats in the modeling group ［bilater hippocampus injected with 1 μL amyloid-beta protein 1-42 （Aβ_1-42） solution for the modeling of AD］. Fifty successfully modeled rats were selected and randomly assigned into model， donepezil hydrochloride （0.45 mg·kg^-1）， and high-， medium-， and low-dose （15.30， 7.65， 3.82 g·kg^-1） Hei Xiaoyaosan groups. Rats were administrated with corresponding agents by gavage once a day for 42 days. Terminal-deoxynucleoitidyl transferase-mediated nick end labeling （TUNEL） was employed to observe the apoptosis of neurons in the cortex and hippocampus， and immunohistochemistry （IHC） was used to detect the expression of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to determine the mRNA levels of Fas， FasL， and Fas-associated protein with death domain （Fadd）. Western blot was used to determine the protein levels of Fas， FasL， Fadd， Caspase-3， cleved Caspase-3， Caspase-8， and cleved Caspase-8. ResultCompared with the blank group and sham group， the model group showed increased apoptosis rate in the cortex and hippocampus （P<0.01）， elevated Bax level （P<0.01）， lowered Bcl-2 level （P<0.01）， up-regulated mRNA levels of Fas， FasL， and Fadd in the hippocampus （P<0.01）， and up-regulated protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.01）. Compared with the model group， donepezil hydrochloride and Hei Xiaoyaosan at high and medium doses decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bax level （P<0.01）， elevated the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of Fas， FasL， and Fadd and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05， P<0.01） in the hippocampus. Low-dose Hei Xiaoyaosan decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of FasL and Fadd （P<0.05） and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05） in the hippocampus. ConclusionHei Xiaoyaosan can protect neurons in the cortex and hippocampus of AD rats by inhibiting the apoptosis mediated by the Fas/FasL/Caspase-8/Caspase-3 signaling pathway.

ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in modulating the synaptic plasticity in APP/PS1 mice by regulating the cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA）/N-methyl-D-aspartate receptor （NMDAR） signaling pathway. MethodTwelve 4-month-old male C57BL/6J mice were selected as the blank control group， and 60 4-month-old male APP/PS1 double transgenic mice were randomized into model， KW-6002 （adenosine receptor antagonist， 3 mg·kg^-1）， and high-， medium-， and low-dose （22.10， 11.05， 5.53 g·kg^-1， respectively） Hei Xiaoyaosan groups， with 12 mice in each group. Mice were administrated with corresponding drugs for 90 days. Transmission electron microscopy was employed to observe the synaptic ultrastructure of hippocampal neurons， and Golgi staining was used to observe the dendritic spine density of neurons in hippocampal CA1 region. Western blot was employed to measure the protein levels of cAMP， PKA， N-methyl-D-aspartate receptors 1， 2A， and 2B （NR1， NR2A， and NR2B， respectively）， postsynaptic density protein 95 （PSD95）， and synapsin 1 （SYN1）. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was performed to determine the mRNA levels of cAMP， PKA， and NR1. Enzyme-linked immunosorbent assay was employed to determine the content of interleukin-12 （IL-12） and interleukin-4 （IL-4） in the hippocampus. ResultCompared with the blank group， the model group showed blurred boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region， reduced and scattered synaptic vesicles， and decreased density of postsynaptic membrane， and irregular， disarranged， and loosened dendritic spines of neurons in hippocampal CA1 region （P<0.01）. In addition， the model group presented down-regulated protein levels of cAMP， PKA， NR1， NR2A， NR2B， PSD95， and SYN1 and mRNA levels of cAMP， PKA， and NR1， elevated IL-12 level， and lowered IL-4 level in the hippocampus （P<0.01）. Compared with the model group， the drug intervention groups showed clear and intact boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region， increased synaptic vesicles with dense arrangement， increased density of postsynaptic membrane， and improved morphology， arrangement， and density of neuronal dendritic spines （P<0.05， P<0.01）. In addition， the drug interventions up-regulated the protein levels of cAMP， PKA， NR1， NR2A， NR2B， PSD95， and SYN1 （P<0.05，P<0.01） and mRNA levels of cAMP， PKA， and NR1 （P<0.01）， lowered the IL-12 level （P<0.01）， and elevated the IL-4 level （P<0.01） in the hippocampus. ConclusionHei Xiaoyaosan can improve the structure and morphology of hippocampal neurons in APP/PS1 mice by activating the cAMP/PKA/NMDAR signaling pathway and repairing synaptic plasticity.