[Potential molecular mechanism of Banxia Xiexin Decoction in treatment of colon cancer based on network pharmacology and molecular docking technology].

The aim of this paper was to explore the potential molecular mechanism of Banxia Xiexin Decoction in the treatment of colon cancer through pharmacology network and molecular docking methods. The chemical constituents and action targets of 7 herbs from Banxia Xiexin Decoction were collected by using TCMSP database,Chinese Pharmacopoeia and literatures consultation. GeneCards database was used to predict the potential targets of colon cancer. GO biological process analysis and KEGG pathway enrichment analysis of the disease and drug intersection targets were carried out through DAVID database. "Component-target-pathway" network and protein-protein interaction(PPI) network were construction by using Cytoscape and STRING database,and then the core components and targets of Banxia Xiexin Decoction in the treatment of colon cancer were selected according to the topological parameters. Finally, Autodock Vina was used to realize the molecular docking of core components and key targets. The prediction results showed that there were 190 active compounds and 324 corresponding targets for Banxia Xiexin Decoction,involving 74 potential targets for colon cancer. Cytoscape topology analysis revealed 11 key targets such as STAT3,TP53,AKT1,TNF,IL6 and SRC, as well as 10 core components such as quercetin,ß-sitosterol,baicalein,berberine,and 6-gingerol.In bioinformatics enrichment analysis, 679 GO terms and 106 KEGG pathways were obtained, mainly involving PI3 K-AKT signaling pathway,TNF signaling pathway and TP53 signaling pathway. The results of molecular docking showed that baicalein,berberine,licochalcone A and 6-gingerol had a high affinity with SRC,STAT3,TNF and IL6. The results suggested that Banxia Xiexin Decoction could play an anti-colon cancer effect by inhibiting cell proliferation, regulating cell cycle, inducing apoptosis and anti-inflammatory function. The study revealed the multi-components,multi-targets and multi-pathways molecular mechanism of Banxia Xiexin Decoction,which could provide scientific basis and research ideas for the clinical application of Banxia Xiexin Decoction and the treatment of colon cancer with compound Chinese medicines.

Correlation of the predisposition of Chinese children to cerebral palsy with nucleotide variation in pri-miR-124 that alters the non-canonical apoptosis pathway.

Cerebral palsy is a group of non-progressive motor impairment syndromes caused by brain lesions during development. Herein, we investigated the relationship between nucleotide variations in a miRNA coding region and the predisposition of Chinese children to cerebral palsy. A total of 233 CP patients and 256 healthy participants were enrolled, and 60 children were selected from each group for plasma miRNA detection. We screened the coding regions of pri-miR-124-1, -2, and -3 using PCR and sequencing. The expression of miR-124 was determined by qRT-PCR. Luciferase assays and Western blots were used to confirm the regulation of target genes by miR-124. The function of miR-124 was further identified in SH-SY5Y cells by detecting cell viability and apoptosis. We revealed that the rare alleles T of rs3802169 and G of rs191727850 were found to be associated with an increased risk of cerebral palsy (OR=3.71, 95% CI 1.74-7.92 and OR=2.18, 95% CI 1.36-3.49, respectively). We further showed that the levels of mature miR-124 were down-regulated by the C-to-T variation in vitro. More importantly, the reduction of miR-124 resulting from the C-to-T change led to the less-efficient inhibition of the target genes ITGB1, LAMC1 and BECN1, which may play important roles during the development of the nervous system. Meanwhile, the reduction in the expression of miR-124 was also related to the increased nuclear translocation of apoptosis-inducing factor (AIF) under oxidative stress, thereby inducing more cell apoptosis. Our results suggest that one functional polymorphism in pri-miR-124-1 might contribute to the genetic predisposition of Chinese children to cerebral palsy by disrupting the production of miR-124, which consequently interfered in the expression and function of the target genes of miR-124.

[Effect of intensive insulin therapy on apoptosis-related ligands in serum in rats with severe scald].

OBJECTIVE: To investigate changes in apoptosis-related ligands in serum in rats with severe scald and the effect of intensive insulin therapy on the changes. METHODS: One hundred and fifty Wistar rats were randomly divided into 3 groups: sham burn (SB), scald (S) and treatment (T) groups. Rats in S and T groups were inflicted with 40% TBSA full-thickness burn, followed by intraperitoneal injection with 40 mL/kg of isotonic saline for resuscitation. Rats in T group were subcutaneously injected insulin in a dose of 0.25 U/100 g 24 hours after burn injury, and every 12 hours for 5 days (0.25, 0.50, 0.75, 1.00, 1.25 U/100 g each day, respectively) to control the level of blood glucose between 3 and 6 mmol/L. Rats in SB group were sham scalded at 37 degrees C without resuscitation. Blood was drawn from abdominal aorta on 1, 4, 7, 10, 14 post burn day (PBD) for determination of serum levels of TNF-alpha, soluble Fas ligand (sFasL) and soluble Fas receptor (sFas) by enzyme-linked immunosorbent assay (ELISA), and insulin by radioimmunity assay (RIA). RESULTS: The serum level of TNF-alpha in S group peaked on 1 PBD (30.9 +/- 8.7) ng/L, which showed statistically significant difference when compared with that of SB and T groups (12.7 +/- 2.8) ng/L, (16.8 +/- 4.7) ng/L, respectively, P < 0.01), then lowered gradually to become similar to that of SB group on 7 PBD. The level of TNF-alpha in T group increased gradually, but was obviously lower than that of S group on 1, 4, 7 PBD (P < 0.01). The level of sFasL in S (on 7-14 PBD) and T (4-10 PBD) groups was significantly higher than that in SB group (P < 0.05), then lowered to normal level. The levels of sFas on 4-10 PBD in T group were obviously higher than that in S and SB group (P < 0.05). Ratio of sFasL to sFas in serum of S group was higher than that in SB group on 7, 10 PBD, which was higher than that in T group on 7 PBD (P < 0.05). There was significant decrease in serum level of insulin in S group compared with that of SB group on 4-10 PBD (P < 0.05). The level of insulin in T group increased on 1 PBD, peaked on 4 PBD (327 +/- 15 microU/mL), which was significantly higher than that in SB and S groups (42 +/- 15, 28 +/- 10 microU/mL, respectively, P < 0.01), then decreased gradually to normal level. CONCLUSIONS: Insulin may inhibit apoptosis after burn by down-regulating secretion of apoptotic ligands.

[Preliminary study of skeletal muscle apoptosis after severe thermal injury in rats].

OBJECTIVE: To investigate changes in skeletal muscle apoptosis after a severe thermal injury in rats. METHODS: One hundred Wistar rats were randomly divided into two groups: sham thermal injury group and severe thermal injury group. They were subdivided into 1, 4, 7, 10, 14 days post-injury with 10 rats in each subgroup. Rats in severe thermal injury group were subjected to a 40% total body surface area full-thickness scald injury. Both weight and tibialis anterior (TA) mass of rats were weighed on 1, 4, 7, 10, 14 days post-injury. Electron microscope was used for observing ultrastructural changes in skeletal muscle, including apoptosis. Tissues of tibialis anterior from burn and sham burn animals were then examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunohistochemistry. RESULTS: Compared with sham thermal injury group, body weight and TA mass of rats were decreased from first day on, and it dropped to the lowest level at 4 days (P<0.05 and P<0.01), and started to regain from 7 days on (all P<0.01). Electron micrographs showed condensation of chromatin around the periphery of the nucleus, blebbing of the sarcolemma, and free of myofibrils near myonuclei in a large area in skeletal muscle of thermally injured rats. Sporadic TUNEL positive myonuclei were also seen under light microscope in skeletal muscle in thermal injury group. There were no characteristic signs of apoptosis in skeletal muscle in rats of sham group. CONCLUSION: There are skeletal muscle apoptosis after severe thermal injury. It may contribute to atrophy of skeletal muscle after burn injury in rats.

Regulation of signaling pathways downstream of IGF-I/insulin by androgen in skeletal muscle of glucocorticoid-treated rats.

BACKGROUND: The mechanisms by which androgens ameliorate glucocorticoid-induced muscle wasting are still under investigation. In the present study, we tested the hypothesis that androgen's effects in reversing muscle wasting are related to activating the signaling pathways downstream of insulin-like growth factor-1 (IGF-I)/insulin. METHODS: Forty female Sprague-Dawley rats were randomly divided into four groups: control group, dexamethasone (DEX) group, testosterone (TES) group, and TES + DEX group. Each group was injected with saline or DEX (0.1 mg/100 g/d) for 10 days and sesame oil or TES (0.5 mg/100 g/d) for 13 days. Several downstream targets of IGF-I/insulin in skeletal muscle including protein kinase B (Akt), p70 ribosomal protein S6 kinase (p70S6K), and glycogen synthase kinase-3beta (GSK-3beta) that are associated with protein synthesis were examined. Two proteolysis-related ubiquitin E3-ligases, muscle atrophy F-box, and muscle RING finger-1 that are also regulated by IGF-I/insulin were also assessed. RESULTS: TES attenuated gastrocnemius muscle atrophy induced by DEX. TES prevented the DEX-induced decrease of IGF-I expression in gastrocnemius muscle, but not in serum. TES ameliorated DEX-induced dephosphorylation of Akt and p70S6K and promoted the phosphorylation of GSK-3beta in gastrocnemius muscle. The total amount of Akt, p70S6K, or GSK-3beta proteins was not changed among these groups. TES did not show any effects on the DEX-induced upregulation of muscle atrophy F-box, and muscle RING finger-1 mRNA in gastrocnemius muscle. CONCLUSION: This findings suggest that the effects of TES in reversing DEX-induced muscle atrophy are related to signaling pathways downstream of IGF-I/insulin that are associated with protein synthesis.

[Repairing of lower eyelid ectropion with expanded flap].

OBJECTIVE: To investigate the effect of reconstruction of lower eyelid ectropion with expanded flap. METHODS: Fourty patients with lower eyelid ectropion were reconstructed using tissue expander. The volume of the smallest expander was 30 ml, and that of the biggest one was 150 ml. The expand time was from 2-months to 3-months, then advancement or transposition flaps were created and employed in the defected lesion where the scar was removed just in one operation. RESULTS: All patients have been followed up for 2-year with satisfactory results and no recurrences was appearance. CONCLUSIONS: Application of expander reasonable may get satisfactory result in reconstruction of lower eyelid ectropion. The incision in donor site is hidden and the symptom seldom recurs.

[Expression of ephrin-B3 mRNA and cellular apoptosis in the brain of neonatal rats with periventricular leukomalacia].

OBJECTIVE: To study the changes of ephrin-B3 mRNA expression and cellular apoptosis in the brain of neonatal rats with periventricular leukomalacia (PVL) and to explore the possible role of ephrin-B3 in the pathogenesis of PVL. METHODS: Two-day-old SD rats were randomly assigned to two groups: PVL and control. PVL model was prepared by right common carotid artery ligation followed by 4-hr 6% oxygen exposure. The control group, without ligation of the artery and hypoxia treatment, was sham operated. The rats were then sacrificed and brain tissues were collected at 0, 8, 24, 48 and 72 hrs and at 7 days of hypoxic-ischemia (HI). Hematoxylin and eosin staining was used for pathologic studies. Real time RT-PCR was applied to detect brain ephrin-B3 mRNA expression. DAPI staining was applied to detect neuronal apoptosis. RESULTS: The brain ephrin-B3 mRNA expression increased significantly in the PVL group at 8, 24, 48 and 72 hrs and at 7 days of HI compared with that of the control group (P < 0.05). The apoptotic cells in the brain of the PVL group were significantly more than that of the control group at 8, 24, 48 and 72 hrs of HI (P < 0.05). CONCLUSIONS: ephrin-B3 mRNA expression and cellular apoptosis in the brain increased significantly in neonatal rats with PVL, which suggests that ephrin-B3 may participate in the pathogenesis of PVL in neonatal rats.

[Changes in the expression of rat hepatic lipid metabolism related genes in response to burn wound sepsis as assessed by DNA arrays].

OBJECTIVE: To examine the expression profile of lipid metabolism-related genes in liver of scalded rats with wound sepsis, and to analyze its significance. METHODS: Sixty male Wistar rats with 30% TBSA full-thickness scald wound on the back were enrolled in the study and randomly divided into wound sepsis group (n = 30) and control group (n = 30, with scald). Pseudomonas aeruginosa was inoculated to the wounds in sepsis group. Corresponding indices were determined to verify the diagnosis of wound sepsis. The rats were sacrificed and fresh liver tissues were obtained at 96 post-scald hours (PSH). Total RNA of liver was isolated with Trizol and the different expression of lipid metabolism related genes in response to burn wound sepsis was assessed by DNA microarray. RESULTS: By comparing expression profile of the two groups, totally 47 genes were observed to be differentially expressed in rat hepatic tissues, among them 9 genes were related to lipid metabolism. Among them, those which were upregulated were genes in relation with transportation and activation of fatty acid, and those downregulated were genes related in providing energy for fatty acid oxidation in mitochondria. CONCLUSION: The occurrence of wound sepsis in scalded rats can induce changes in the expression of hepatic lipid metabolism related genes in hepatic tissues, and aggravate

[Effect of ubiquitin-proteasome pathway on inflammatory reaction in intestine and its barrier function in rats with postburn sepsis].

OBJECTIVE: To study the effect of ubiquitin-proteasome pathway inhibition on intestinal nuclear factor-KappaB (NF-KappaB) activity and tumor necrosis factor-alpha (TNF-alpha) release as well as plasma diamine oxidase (DAO) activity in rats with postburn sepsis. METHODS: Rats were subjected to 30% total body surface area (TBSA) full-thickness scald injury, followed by intraperitoneal injection of lipopolysaccharide (LPS) to mimic postburn sepsis. Sixty Wistar rats were randomly divided into normal control group, sepsis group, sepsis with proteasome inhibitor N-Acetyl leucinyl leucinyl norleucinal (ALLN) treatment group and sepsis with NF-KappaB inhibitor pyrrolidine dithiocarbamate (PDTC) treatment group. NF-KappaB activity, TNF-alpha protein content, and plasma DAO activity were determined by electrophoretic mobility shift assay (EMSA), enzyme-linked immunosorbent assay (ELISA), and spectrophotometric method, respectively. RESULTS: The results showed that NF-KappaB activity was markedly activated and reached its peak 1 hour after scalding and injection of LPS in each group (all P<0.01), then reduced gradually. Both ALLN and PDTC could decrease intestinal NF-KappaB activity at 1 hour and 2 hours after injury. TNF-alpha release was reduced by ALLN at 1 hour after injury (P<0.01). Plasma DAO activity was significantly elevated after scalding and injection of LPS (P<0.01). Pretreatment with PDTC or ALLN could not lower the activity of DAO. CONCLUSION: The results suggest that early treatment with inhibitor of ubiquitin-proteasome pathway might decrease the intestinal inflammatory reaction, but exert no effect on intestinal barrier function in rats with postburn sepsis.

[The influence of lipopolysaccharide on adipose metabolism in liver during shock stage of scalded rats].

OBJECTIVE: To investigate the influence of lipopolysaccharide (LPS) on adipose metabolism in liver during shock stage of scalded rats. METHODS: Sixty adult Wistar rats were inflicted with 30% TBSA full thickness scald and were randomly divided into 3 groups: i. e. sham group (control, n = 20), simple scald group [(n = 20) and LPS group (n = 20, with intra-peritoneal injection of 3.0 mg/kg LPS at 2 postscald hour (PSH)]. The contents of LPS, tumor necrosis factor alpha (TNF-alpha), free fatty acids (FFA) in plasma and adenosine triphosphate (ATP), triglyceride (TG), malonaldehyde (MDA) in liver in each group were determined at 24 and 48 PSH. The histological changes in hepatic tissue in each group were also observed. RESULTS: The plasma contents of FFA in LPS group at 24 and 48 PSH were 2.3 +/- 0.3 mmol/L and 2.5 +/- 0.4 mmol/L, respectively, which were obviously higher than those in control (0.4 +/- 0.3 mmol/L, 0.5 +/- 0.3 mmol/L) and scald (0.9 +/- 0.3, 1.2 +/- 0.5 mmol/L, P <0.01) groups. Meanwhile, there was obvious difference in the contents of TG and ATP in liver between LPS group (TG: 530 +/- 30 mmol/g, ATP: 1.7 +/- 0.5 micromol/g) and scald group (TG: 242 +/- 27 mmol/g, ATP: 6.0 +/- 2.4 micromol/g, P < 0.01). Pathological examination revealed that adipose denaturalization and injury to mitochondria in hepatocytes in scald group were significantly milder than those in LPS group. The morphology of hepatocyte in control group appeared normal. CONCLUSION: LPS challenge to burn subjects could induce impairment in utilizing fat derived energy, and it would aggravate adipose denaturalization in the liver.

[The role of tyrosine phosphorylation proteins in procollagen gene expression of fibroblasts in wound healing].

OBJECTIVE: To investigate the role of adhesion between fibronectin and fibroblasts in wound healing as well as tyrosine phosphorylation proteins in procollagen mRNA expression. METHODS: The level of proalpha1 (I) mRNA and tyrosine phosphorylation protein were detected employing the techniques of RT-PCR and immunoblotting. After inhibition of tyrosine kinases, herbimycin A was added to the medium to block the pathway of tyrosine phosphorylation, the changes of procollagen mRNA and tyrosine phosphorylation proteins were further investigated. RESULTS: The adhesion between fibroblasts and fibronectin in wound healing not only induced the production of 98kd and 65kd tyrosine phosphorylation protein, but also enhanced obviously the expression of procollagen alpha1 (I) mRNA. When the pathway of tyrosine phosphorylation was blocked, the level of procollagen alpha1 (I) mRNA lowered remarkably, accompanied by the decrease of 98kd, 65kd tyrosine phosphorylation proteins. CONCLUSION: The adhesion between fibronectin and fibroblasts plays an important role in expression increase of procollagen mRNA during wound healing, in the process of which tyrosine phosphorylation is a key step.

[The treatment of deformity of axillary scar contracture after burns].

OBJECTIVE: To explore the best methods to repair the deformity of axillary scar contracture after burns. METHODS: Ninety cases in 78 patients with axillary scar contracture after burns from January 1998 to January 2002 were analyzed. According to the severity of the deformity and its influence on the function of the shoulder joint. 46 cases suffered from mild degree axillary scar contracture, 26 cases with moderate and 18 with severe degree. The deformities of axillary scar contracture were repaired by Z plasty (18 cases), five-flap plasty (14 cases), skin graft (23 cases), Z plasty and skin graft (14 cases), transfer of scapular skin flaps (5 cases), lateral throatic skin flaps (4 cases) and scar flaps (12 cases), respectively. Exopexy, anti-scar drug and functional exercises were applied postoperationally. RESULTS: All the flaps were survived with first intention, except for necrosis of the split skin graft occurring in 4 cases. The function and configuration in all the cases were satisfactory after 6 months to 4 years follow-up. CONCLUSION: Appropriate methods should be chosen to restore the function and configuration of the shoulder joint and improve patients' living condition according to the size, degree of the deformity of axillary scar contracture after burns.

[Defensin-5 and Matrilysin mRNA expression in the intestine of scalded rats and its relation to bacterial translocation].

OBJECTIVE: To investigate the effect of burn injury on the expression of rat defensin-5 (RD-5) and Matrilysin mRNA in the intestine of scalded rats and to determine its relation to bacterial translocation. METHODS: Thirty-two Wistar rats were divided randomly into sham injury group (n=8) and burn group (n=24). Rats of the burn group were subjected to 30% total body surface area (TBSA) full-thickness scald injury. At 8, 24 and 72 hours after the injury, the animals (n=8 at each time point) were sacrificed and ileum was sampled for the determination of the expression of RD-5 mRNA and Matrilysin mRNA with reverse transcription-polymerase chain reaction (RT-PCR) technique. Cultures of mesenteric lymph nodes (MLN), liver, spleen and lung were carried out at 24 hours after the injury to assess the rate of bacterial translocation to visceral organs,and the morphological changes in ileal Paneth cells were observed. RESULTS: The expression of RD-5 mRNA was significantly up-regulated at 8 hours when compared with control group. It was down-regulated sharply at 24 hours but was still higher than control group, and at 72 hours it was down-regulated to a level lower than control. On the contrary, the expression of Matrilysin mRNA was up-regulated continuously to 72 hours, and it was significantly higher than control. Bacterial translocation rate was significantly higher in rats of the burn group at 24 hours (58.3%) than in control group (8.3%, P<0.01), but there were no obvious morphological changes in ileal Paneth cells at the same time point. CONCLUSION: The expressions of both RD-5 mRNA and Matrilysin Mrna are up-regulated in the early stage after the injury, but their time phases are different. The phenomenon might imply that there is a host protective response against bacterial translocation.