Objective:To explore the expression pattern of aryl hydrocarbon receptor (AhR) in mice peritoneal macrophages (PMs) after major trauma and analyze the effects of enhanced AhR expression on the inflammatory cytokine level and bactericidal ability after trauma.Methods:The experimental study method was used. Forty 6-8-week-old male C57BL/6J mice (the same mouse age, sex, and strain below) were divided into control group, post trauma hour (PTH) 2 group, PTH 6 group, and PTH 12 group according to the random number table (the same grouping method below), with 10 mice in each group. Mice in the latter 3 groups were constructed as severe trauma model with fracture+blood loss, while mice in control group were left untreated. The primary PMs (the same cells below) were extracted from the mice in control group, PTH 2 group, PTH 6 group, and PTH 12 group when uninjured or at PTH 2, 6, and 12, respectively. Then the protein and mRNA expressions of AhR were detected by Western blotting and real-time fluorescence quantitative reverse transcription polymerase chain reaction, respectively, and the gene expressions of AhR signaling pathway related molecules were analyzed by transcriptome sequencing. Twenty mice were divided into control group and PTH 6 group, with 10 mice in each group, and the PMs were extracted. The level of ubiquitin of AhR was detected by immunoprecipitation. Twelve mice were divided into dimethyl sulfoxide (DMSO) alone group, PTH 6+DMSO group, MG-132 alone group, and PTH 6+MG-132 group, with 3 mice in each group. After the corresponding treatment, PMs were extracted, and the protein expression of AhR was detected by Western blotting. Twenty mice were constructed as PTH 6 model. Then, the PMs were extracted and divided into empty negative control adenovirus (Ad-NC) group and AhR overexpression adenovirus (Ad-AhR) group. The protein expression of AhR was detected by Western blotting at 36 h after some PMs were transfected with the corresponding adenovirus. The rest cells in Ad-NC group were divided into Ad-NC alone group and Ad-NC+endotoxin/lipopolysaccharide (LPS) group, and the rest cells in Ad-AhR group were divided into Ad-AhR alone group and Ad-AhR+LPS group. The expressions of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in the cell supernatant were detected by enzyme-linked immunosorbent assay at 12 h after the corresponding treatment ( n=6). Twenty mice were obtained to extract PMs. The cells were divided into control+Ad-NC group, PTH 6+Ad-NC group, control+Ad-AhR group, and PTH 6+Ad-AhR group, and the intracellular bacterial load was detected by plate spread method after the corresponding treatment ( n=6). Data were statistically analyzed with one-way analysis of variance, least significant difference test, analysis of variance for factorial design, and independent sample t test. Results:Compared with 1.16±0.28 of control group, the protein expressions of AhR in PMs in PTH 2 group (0.59±0.14), PTH 6 group (0.72±0.16), and PTH 12 group (0.71±0.17) were all significantly decreased ( P<0.05). The overall comparison of the difference of AhR mRNA expression in PMs among control group, PTH 2 group, PTH 6 group, and PTH 12 group showed no statistical significance ( P>0.05). The AhR signaling pathway related molecules included AhR, AhR inhibitor, cytochrome P450 family member 1b1, cytochrome P450 family member 11a1, heat shock protein 90, aryl hydrocarbon receptor-interaction protein, and heat shock protein 70 interaction protein. The heat shock protein 90 expression of PMs in PTH 2 group was higher than that in control group, while the expressions of other molecules did not change significantly after trauma. Compared with that in control group, the level of ubiquitin of AhR in PMs in PTH 6 group was increased. Compared with that in DMSO alone group, the protein expression of AhR in PMs in PTH 6+DMSO group was decreased, while that in PMs in MG-132 alone group had no significant change. Compared with that in PTH 6+DMSO group, the protein expression of AhR in PMs in PTH 6+MG-132 group was up-regulated. At transfection hour 36, compared with that in Ad-NC group, the protein expression of AhR in PMs in Ad-AhR group was increased. At treatment hour 12, compared with those in Ad-NC+LPS group, the expressions of IL-6 and TNF-α in PM supernatant of Ad-AhR+LPS group were significantly decreased (with t values of 4.80 and 3.82, respectively, P<0.05). The number of intracellular bacteria of 1×10 6 PMs in control+Ad-NC group, PTH 6+Ad-NC group, control+Ad-AhR group, and PTH 6+Ad-AhR group was (3.0±1.8), (41.8±10.2), (1.8±1.2), and (24.2±6.3) colony forming unit, respectively. Compared with that in PTH 6+Ad-NC group, the number of intracellular bacteria of PMs in PTH 6+Ad-AhR group was significantly decreased ( t=3.61, P<0.05). Conclusions:Ubiquitin degradation of AhR in PMs of mice after major trauma results in decreased protein expression of AhR. Increasing the expression of AhR in post-traumatic macrophages can reduce the expressions of LPS-induced inflammatory cytokines IL-6 and TNF-α, and improve the bactericidal ability of macrophages after trauma.

Abdomen/diagnostic imaging* ; Artificial Intelligence ; Body Composition ; Magnetic Resonance Imaging ; Practice Guidelines as Topic

This study was aimed to explore the in vitro antibacterial,anti-inflammatory and antitussive effects of water extract of Styrax japonicus leaves and to verify its concerning efficacies.In vitro antibacterial,anti-inflammatory and antitussive effects were observed by in vitro antibacterial experiment,cotton ball implantation,carrageenan-induced paw edema together with xylene-induced ear edema experiments with rats or mice,and ammonia hydroxide-induced coughing experiment,respectively.The results showed that water extract of Styraxjaponicus leaves can inhibit the growth of Escherichia coli,Klebsiella pneumoniae,Salmonella and Staphylococcus aureus except candida albicans.Its high-dose group can reduce the weight of cotton ball granuloma in rats obviously.And its low-dose and high-dose groups can inhibit carrageenan-induced paw edema apparently.Meanwhile,its low-dose and high-dose groups can improve xylene-induced ear edema distinctively.Its low-dose and high-dose groups can evidently prolong the ammonia hydroxide-induced coughing incubation periods.And all groups can reduce coughing times.It was concluded that the water extract of Styrax japonicus leaves coincided with the relative clinical efficacy of traditional Chinese medicine (TCM) and provided experimental evidences for its clinical application to some degree.

Previous studies have demonstrated that integrin-linked kinases (ILKs) are abundantly expressed in extracellular matrix (ECM) riche dermis, hair follicles, and basal cells of epidermis. ILKs are not only essential for the maintenance of skin structure, but also play important roles in wound healing. ILKs can promote the formation of granulation tissue by stimulating the proliferation of fibroblasts and secretion of ECM, accelerate wound contraction by inducing the differentiation of fibroblasts to myofibroblasts, and boost reepithelization by promoting proliferation, migration, and differentiation of keratinocytes and follicle epidermal stem cells.

OBJECTIVETo investigate the role of integrin-linked kinase (ILK) signaling pathway in the skin lesions and wound healing in diabetic rats.

METHODSThirty-six SD rats were divided into diabetic wound group (D) and non-diabetic wound group (N) according to the random number table, with 18 rats in each group. 10 g/L streptozocin (60 mg/kg) was intraperitoneally injected in rats in group D, while the rats in group N were given same quantity of sodium citrate buffer. Two weeks after successful reproduction of diabetic model of rats in group D, two full-thickness skin of an area of 2 cm × 2 cm was resected on both sides of back of rats in the two groups. Wounds of three rats of each group were photographed and examined on post injury day (PID) 1, 3, 7, 10, 14, and 21, and the wound healing rates were calculated. The non-injured skin and wound tissue (central part) on back of three rats of the rest 15 rats in the two groups were harvested on PID 3, 7, 10, 14, and 21, respectively. Morphology of the non-injured skin tissue was observed with HE staining, and the thickness of full-thickness skin and epidermis were measured. The mRNA expression levels of ILK, protein kinase B (Akt), and glycogen synthase kinase-3β (GSK-3β) in non-injured skin tissue were determined with real-time fluorescent quantitative RT-PCR. The protein expression levels of ILK, Akt, phosphorylated Akt, GSK-3β, and phosphorylated GSK-3β in non-injured skin tissue, and ILK, phosphorylated Akt in wound tissue were assessed with Western blotting. Data were processed with two independent-sample t test, one-way analysis of variance, SNK test and analysis of variance of factorial design.

RESULTS(1) After injury, the wound scabs of rats in group N were dry, and red granulation tissue with no excretion were seen when the scabs fell off, and the wound healed fast. After injury, excretion under the wound scabs of rats in group D was seen, and the scabs easily fell off with exposure of pink granulation tissue with much excretion, and the wounds healed slowly. Except for PID 3, the wound healing rate of rats in group D was significantly lower than that in group N on other PIDs (with t values from 3.858 to 13.738, P<0.05 or P<0.01). (2) On PID 3, the hair follicles and blood vessels in the non-injured skin tissue of rats in group N were rich, and the epidermis was composed of stratified cells in form of basal cells and keratinocyte, and the hair follicles and blood vessels in the non-injured skin tissue of rats in group D were scarce, and the epidermis was nearly composed of one-layer of cells. The thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group N was similar from PID 3 to 21, and the thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group D on PID 3 was respectively (1 074 ± 66) and (15.1 ± 3.8) μm, and they gradually thinned out to (785 ± 122) and (9.7 ± 2.1) μm on PID 21, respectively. The thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group N were significantly thicker than those in group D on each PID (with t values from 4.620 to 23.549, P values below 0.001). (3) From PID 3 to 21, the mRNA expression levels of ILK and Akt in non-injured skin tissue of rats in group D were significantly lower than those in group N (with t values respectively 4.779 and 3.440, P values below 0.05), the mRNA expression levels of GSK-3β in non-injured skin tissue of rats were similar in two groups (t=0.363, P>0.05). (4) From PID 3 to 21, the protein expression levels of ILK, Akt and phosphorylated Akt in non-injured skin tissue of rats in group D were significantly lower than those in group N (with t values from 2.630 to 6.209, P<0.05 or P<0.01); the protein expression levels of GSK-3β in non-injured skin tissue of rats in two groups were similar (t=0.652, P>0.05); the protein expression level of phosphorylated GSK-3β in non-injured skin tissue of rats in group D was significantly higher than that in group N (t=4.131, P<0.001). The protein expression levels of ILK in wound tissue of rats in two groups were similar on each PID (with t values from 0.381 to 2.440, P values above 0.05). Except for PID 3, the protein expression levels of phosphorylated Akt in wound tissue of rats in group N were significantly higher than that in group D on other PIDs (with t values from 4.091 to 20.555, P<0.05 or P<0.01). From PID 3 to 21, the protein expression levels of ILK in wound tissue and non-injured skin tissue of rats in group N were similar (F=2.522, P>0.05), and the protein expression level of phosphorylated Akt in wound tissue was significantly higher than that in non-injured skin tissue (F=117.329, P<0.001); the protein expression levels of ILK in wound tissue and non-injured skin tissue of rats in group D were similar (F=1.337, P>0.05), and the protein expression level of phosphorylated Akt in wound tissue was significantly higher than that in non-injured skin tissue (F=184.120, P<0.001).

CONCLUSIONSThe skin lesion of diabetic rats may be related to the declined expression levels of ILK, Akt and phosphorylated Akt in the ILK signaling pathway. The refractory healing of wound in diabetic rats may be related to the declined expression level of phosphorylated Akt.

Animals ; Diabetes Mellitus, Experimental ; enzymology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Phosphorylation ; Protein-Serine-Threonine Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Skin ; injuries ; Wound Healing

Objective To investigate the effects of gastric bypass on glycometabolism and improvement of islet β cell function and insulin resistance in patients with type 2 diabetes. Methods Eight patients with type 2 diabetes combined with gastric carcinoma who treated with gastric bypass were studied prospectively. Fasting and postprandial plasma glucose levels, fasting and postprandial insulin C-peptide levels, and body mass index (BMI) were measured right before the surgery and at intervals of 1 week, 2 weeks, 1 month and 3 months after the surgery. Glycosylated hemoglobin (HbA1c) levels were measured before and 3 months after the surgery. The outcome of the diabetes after 3 months of the surgery was also monitored. Results Fasting and postprandial plasma glucose levels decreased (P ＜ 0.05) and fasting and postprandial insulin C-peptide levels increased (P ＜ 0.05) after the surgery. HbA1c levels also decreased (P ＜ 0.05) after 3 months of the surgery. There was no significant change of BMI at all intervals after the surgery(P＞ 0.05). All of the 8 patients reached the total effective standard and 6 patients reached the clinical remission standard after 3 months of the surgery. Conclusions It suggests that gastric bypass can significantly lower plasma glucose levels in type 2 diabetes, which does not depend on the loss of weight. The control of plasma glucose by gastric bypass may be due to the improvement of islet β cell function and increasing secretion of endogenous insulin.

Objective To explore the method of isolation, identification and in vitro differentiation of islet stem cells in neonatal rat. Methods The whole pancreata of neonatal rats were digested with collagenase, then,under the (condition) of pH 7.4～7.6,the digested tissue fragments were cultured with serum and serum-free RPMI 1640 (( bFGF), EGF, N2).The whole formation process of new islet cell-like cell masses was examined.The insulin (release) test was used to detect islet function. The expression of nestin was tested by immunocytochemistry. Results The nestin positive cells can be found within 36 h of culture of the digested pancreatic fragments.After addition of bFGF,ECF,N2,nestin positive cells proliferated fast and formed new islets-like cell masses after 18～24 d, and (insulin)ssion could be confirmed. Conclusions The nestin positive cells of pancreatic cells possess the character of islet stem cells, and can form islet like cell masses through culture in vitro.

Objective The evaluation and possibility of percutaneous fenestration for ischemic complications of aortic dissection were disccused. Methods A male patient with aortic dissetion (type: DeBakey Ⅲb) accompanyed by lower extremities pararises was undertaken percutaneous fenestration. Results Both the lower leg's blood flow was recovered with symptom free. Bilateral femoral and dorsal foot pulsations could be feeled. Conclusion Percutaneous fenestration for ischemic complications of aortic dissection is a safe and effective management, but should be done earlier.

Objective To explore the isolation, culture of nestin positive cells of the pancreas in newborn rats. Methods The whole pancreas of neonatal rats were digested with collagenase, followed by incubation under pH 7. 6 serum RPMI 1640 for 24 - 36 h and then under pH 7. 4 serum free RPMI 1640 (bFGF.EGF 20 ng/ml.1% N2) for 18-24 d. The expression of insulin.glucagon and nestin were detected by immunofluorescence technique. Nestin.CK19 were detected by RT-PCR. Resides Cells attached to the surface of plates after 24 h incubation under pH 7. 6 condition, and islet-like masses were obtained after 18 -24 d incubation. A monolayer of cells grew out after 24 h of passage of islet-like masses. Nestin positive cells was detected after 24 - 36 h incubation, with no expression of insulin and glucagon. Positive cells of insulin and glucagon were detected in islet-like masses after 24 h passage. Nestin positive cells were detected by RT-PCR in islet-like masses after 24 h passage, but no CK19. Conclusion Insulin and glucagon were expressed in islet-like masses after passage. Nestin positive cells in the pancreas of neonatal rats possessed character of islet stem cells.

Forty-eight cases of atrial septum secundum defect were repaired and followed up with M-mode echocardiography postoperatively for three months to three yaers respectively.All the cases showed a significant reduction of the inner diameter of the right ventricle after operation,and in 23% of the cases, the inner diameter even returned to the normal range. In 92% of the acses, the intervetricular septal paradoxical motion resumed the normal pattern. In most cases, the inner diameter of the left ventricle increased markedly.In 11 cases with repeated follow-up studies, it was found that three months after operation, the interventricular septal paradoxical motion became normal in 7 cases and the motion flattened in three cases. In 18 cases with a 3-year follow-up history, it was found that the recovery of the heart condition showed no relation to the age of the patient. This is likely to suggest that the age factor exerts no influence on the postoperative recovery of the cases having the atrial septal defect repaired.