Objective　To investigate the expression of protein tyrosine kinase 2 (MFGE8) in patients with ischemic brain injury (IBI) and its regulation on macrophage polarization.Methods　ELISA was used to detect the expression of MFGE8 protein in peripheral blood of patients with ischemic stroke and middle cerebral artery occlusion (MCAO) rat models;IF was used to detect the localization and expression of MFGE8 in brain;BV-2 microglia was treated with the culture supernatant of N2a neuronal cells (Mfge8CA) stably transformed with Mfge8.The polarization ratio of macrophages was detected by flow cytometry.Western blot detection Mfge8,αv/β 3-integrin,FAK,NF-κB.ERK1/2,JNK1/2,P38,PI3K,AKT,mTOR protein expression.Results　The relative content of MFGE8 in peripheral blood of IBI patients and MCAO model rats was significantly lower than that of the control group (Ctrl,P=0.0446,P=0.0259).MFGE8 was highly co-localized with neuron cell marker (NeuN);the proportion of M1 type (CD45+F4/80+iNOS+Arginase1-) macrophages in the brain tissue of MCAO model was significantly higher than that in the Ctrl (P=0.0004).The proportion of M2 type (CD45+F4/80+iNOS-Arginase1+) macrophages was significantly lower than that of the Ctrl (P<0.0001).The proportion of M1 macrophages of BV-2 microglia after supernatant of N2a (mfge8CA) treatment was significantly lower than that of Wild type (WT,P=0.0230).The proportion of M2 macrophages was significantly higher (P<0.0001).The protein expressions of α v/β3-integrin,FAK,p-P85,P85,p-AKT (Ser473),p-mTOR (Ser2481) and p-mTOR (Ser2488) in BV-2 microglia after supernatant of N2a (mfge8CA) treatment were significantly higher than those in WT group.The expression of p-P65 protein was significantly lower than that in WT group.Conclusion　MFGE8 is highly expressed in peripheral blood of patients with IBI.MFGE8 derived from neuronal cells may promote BV-2 microglia M2 macrophages polarization by activating PI3K/AKT/mTOR signals,and inhibit the polarization of M1 macrophages.

Objective To evaluate the effect of parathyroid hormone (PTH) on the expression of connective tissue growth factor (CTGF) in human renal tubular epithelial cells, and to explore the role of MAPK signaling pathway. Methods Real time RT-PCR, Western blot, and reporter gene assay were employed to detect PTH-induced CTGF expression in HK-2 cells. Inhibitors (PD98059 and U0126) of MAPK signaling pathway were used to confirm involved signal pathway. Results HK-2 cells had basic expression level of CTGF mRNA and protein, which were increased significantly after treatment with PTH. The luciferase activity was up-regulated to a higher level as compared with control group after treatment with 10-10 mol/L PTH for 12 h [(1.8884±0.0780) vs (0.9891±0.0300) A, P<0.01]. Moreover, a small amount of p-ERK1/2 was detected in normal HK-2 cells, but it was increased significantly in response to PTH activation, most remarkably when treated with 10-10 mol/L PTH for 30 min. Inhibitors of MAPK signaling pathway, PD98059 and U0126, noticeably inhibited the expression of CTGF mRNA and protein as well as gene promoters in HK-2 cells. Conclusion PTH can induce higher expression of CTGF in HK-2 cells probably via MAPK signaling pathway.

Objective To investigate the effect of parathyroid hormone (PTH) on the transition and connective tissue growth factor (CTGF) expression of human renal proximal tubular epithelial cell line HK-2 . Methods The expression of CTGF mRNA and protein of HK-2 cells were measured by real time RT-PCR and Western blot respectively . The effect of PTH on the phenotypic transformation of HK-2 cells was examined by light microscopy . The expression of α-smooth muscle actin (α-SMA) in HK-2 cells was detected by immunofluorescence . Results Basal level of CTGF mRNA and the protein expression were detected in HK-2 ceils . PTH upregulated the expression of CTGF mRNA and protein with the maximal response at the concentration of 10-10 mol/L and the best stimulating time was at 72 h . After exposure to PTH (10-10tool/L) for 12 hours, the highest level of luciferase activity was 1 .96 fold as compared to control (1 .888±0 .078 vs 0 .989±0 .030, P＜0 .01 ) . Untreated cells showed negligible expression of ±-SMA,whereas ±-SMA expression was significantly increased in cells treated with PTH . Conclusion PTH up-regulates CTGF expression and induces transition of HK-2 cells .

BACKGROUND:Stem cel is a kind of pluripotent cels with self-replication ability, which can differentiate into various cels under certain conditions. Furthermore, stem cels are rich in a variety of growth factors, which can induce the generation of vessels and nerves, and improve the blood supply of lower limbs, thereby achieving the treatment and preventions of lower limb ischemia OBJECTIVE:To summarize and compare the recent achievements in the theory and therapeutic efficacy of stem cels from different sources in the treatment of diabetic foot. METHODS:The first and second authors retrieved PubMed, Sciencedirect and Medline databases for relevant articles published from January 2000 to January 2015. The key words were “diabetic foot, pathogenesis, stem cel therapy” in English. Initialy, 186 articles were retrieved, and finaly 44 articles were included in result analysis. RESULTS AND CONCLUSION:Stem cels can be a new choice for the treatment of diabetic foot. After stem cel therapy, corresponding symptoms have been aleviated, including the generation of new blood vessels and the reshaping of the colateral vessels, the improvement of motor nerve conduction velocity and nerve reflex, the improvement of the sense of skin pain and temperature, and pain relief. It is stil unclear whether alogeneic stem cels are safe or not, but autologous stem cels, especialy bone marrow mesenchymal stem cels, can be better able to repair damaged vessels and nerves and restore the microcirculation of blood supply. Currently, we need to do more basic and clinical researches to solve the folowing problems: to confirm the effectiveness and safety of stem cel therapy for diabetic foot; to identify whether there is a difference in the differentiation and secretory activity between stem cels in diabetic patients and ordinary people; to give ful play to the treatment of diabetic foot.

Laser photocoagulation is one of the important methods for treating retinal diseases, and retinal laser technology continues to advance. For decades, researchers have been striving to find a laser treatment that can minimize tissue damage while achieving optimal results. With low toxicity, low scattering light, strong penetrating power, small compared with the traditional laser damage, light reaction and no pain, the 577 nm subthreshold micropulse laser(SML)turns this goal into reality and ushers in a new era of laser treatment for fundus diseases. This article reviews the concept, mechanism, related parameters and clinical application progress of 577 nm SML in a variety of retinal diseases, aiming to provide references for clinical treatments.

AIM: To investigate the characteristics of changes in corneal epithelial thickness at the early postoperative stage of femtosecond assisted laser in situ keratomileusis(FS-LASIK)and its related influencing factors.METHOD: Retrospective study. A total of 120 patients(240 eyes)of myopia undergoing FS-LASIK from May 2021 to June 2022 were selected. The corneal epithelium thickness in the central area, inner ring area, and outer ring area of patients before and at 1d, 1wk, 1 and 3mo after operation was recorded. The relationship between the variation of corneal epithelium thickness and spherical equivalent, optical zone diameter, depth of cut and cutting ratio was analyzed by Pearson correlation.RESULTS: There was no statistical significance in corneal epithelial thickness in the central area, inner ring area and outer ring area at 1d after FS-LASIK compared with that before operation(P>0.05). At 1wk, 1 and 3mo after surgery, the corneal epithelial thickness in the central area, inner ring area and outer ring area increased compared with that before surgery, and the corneal epithelial thickness in the central area and inner ring area at 1 and 3mo after surgery was greater than that in the outer ring area(all P<0.05). The corneal epithelial thickness in the central, inner and outer ring areas of high myopia patients was thicker than that of low and moderate myopia patients before operation. The corneal epithelial thickness in the central, inner and outer ring areas of high myopia patients was thinner than that of low and moderate myopia patients at 1wk after operation(P<0.05). At 1 and 3mo after operation, the corneal epithelial thickness in the central, inner and outer ring areas of patients with high myopia was greater than that of patients with low and moderate myopia, and the changes of corneal epithelial thickness in the central, inner and outer ring areas were greater than those of patients with low and moderate myopia(P<0.05). The results of Pearson correlation showed that the changes in corneal epithelial thickness in the central and inner ring area were positively correlated with the corneal curvature, depth of cut and cutting ratio at 3mo after surgery, and they were in negative correlation with the age, spherical equivalent and optical zone diameter(P<0.05).CONCLUSION: The corneal epithelial thickness of patients thickened after the FS-LASIK operation, and it was correlated with age, corneal curvature, preoperative depth of cut, cutting ratio, spherical equivalent and the optic zone diameter.

Objective:To investigate the effect of sleeve gastrectomy on blood glucose in obese rats with diabetes mellitus.Methods:Thirty-two 12 week old Goto Kakizaki (GK) diabetic rats were randomly divided into 4 groups: sham operation group (A) , sleeve stomach operation group (B) , sleeve stomach operation+external bile drainage group (C) and sleeve stomach operation + external bile drainage + oleanolic acid group (D) . The changes of fasting blood glucose and blood bile acid were compared before and after operation. The expression level of GLP-1 in serum of each group was detected by ELISA, and the difference of protein expression of bile acid G protein coupled receptor (TGR5) was detected by Western blot.Results:The blood glucose level of group A had no significant change after operation. But blood glucose level in group B and group D was significantly lower than that before operation. Blood glucose in group C was lower than that before operation, but there was no significant difference in comparison. Serum total bile acid level in group A had no significant difference before and after operation. Bile acid level in group B was significantly higher than that before operation, while bile acid level in group C and group D was significantly lower than that before operation. There was no significant difference in the level of serum GLP-1 in group A before and after operation. The level of serum GLP-1 in group B and group D was significantly higher than that before operation. The level of serum GLP-1 in group C was lower than that before operation, but there was no significant difference. The expression of TGR5 protein in terminal ileum in group B and group D was higher than that in group A, but the expression in group C was similar with that in group A.Conclusions:Sleeve gastrectomy has definite hypoglycemic effect on T2DM rats. Bile acid and its related receptor TGR5 should play an important role in the mechanism of sleeve gastrectomy in treatment of diabetes.

Objective:To compare the predictive value of Oxford acute severity of illness score (OASIS) and simplified acute physiology score Ⅱ (SAPSⅡ) for in-hospital mortality in intensive care unit (ICU) patients with sepsis.Methods:A retrospective cohort study was conducted using the data in the Medical Information Mart for Intensive Care-Ⅳ0.4 (MIMIC-Ⅳ 0.4). Based on Sepsis-3 diagnostic criteria, the basic information of ICU adult sepsis patients with infection and sequential organ failure assessment (SOFA) score ≥ 2 within 24 hours of ICU admission admitted for the first time in the database was extracted, including gender, age, vasopressor drugs, sedative drugs, mechanical ventilation, renal replacement therapy, length of ICU stay, OASIS, SAPSⅡ scores, etc. The primary outcome was in-hospital mortality. A receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) was calculated to compare the prognostic value of OASIS score and SAPSⅡ score.Results:A total of 11 098 adult ICU sepsis patients were enrolled in the final analysis, of which 2 320 died and 8 778 survived in hospital, with a mortality of 20.90%. Compared with the survivors, the non-survivors were older [years old: 71 (60, 81) vs. 67 (56, 78)], had longer length of ICU stay [days: 6.95 (3.39, 13.07) vs. 4.23 (2.19, 9.73)] and higher proportions of using vasopressor drugs, sedative drugs, mechanical ventilation and renal replacement therapy [vasopressor drugs: 50.65% (1 175/2 320) vs. 33.05% (2 901/8 778), sedative drugs: 58.53% (1 358/2 320) vs. 48.41% (4 249/8 778), mechanical ventilation: 89.57% (2 078/2 320) vs. 81.66% (7 168/8 778), renal replacement therapy: 11.98% (278/2 320) vs. 6.57% (577/8 778), all P < 0.01]. Moreover, the non-survivors had higher OASIS score [43 (36, 49) vs. 35 (29, 41), P < 0.01] and SAPSⅡ score [49 (40, 60) vs. 38 (31, 47), P < 0.01] as compared with the survivors. ROC curve analysis showed that the AUC of OASIS score and SAPSⅡ score for predicting in-hospital death of ICU patients with sepsis was 0.713 [95% confidence interval (95% CI) was 0.701-0.725] and 0.716 (95% CI was 0.704-0.728), respectively, and the Delong test showed no significant difference in AUC between the two scoring systems ( P > 0.05). Conclusions:OASIS score has a good predictive value for in-hospital mortality in sepsis patients, which is similar to SAPSⅡ score. OASIS score is simpler and has a broader clinical application prospect than SAPSⅡ score.

Objective To investigate anti-cancer effects and mechanism of caffeine on promoting apoptosis of gastric cancer.Methods The viability and proliferation of gastric cancer cells were evaluated by harvesting and counting cells at preset time points after treatment with specific concentrations of caffeine.Flow cytometry was performed to assess cell cycle dynamics and apoptosis.Western blot analysis was used to detect the activity of the above signalling pathway.Results Caffeine treatment significantly suppressed GC cell growth and viability and induced apoptosis by enhancing the Caspase-9/3 pathway.Conclusion Caffeine can effectively promote the apoptosis of gastric cancer cells.

Objective To investigate anti-cancer effects and mechanism of caffeine on promoting apoptosis of gastric cancer.Methods The viability and proliferation of gastric cancer cells were evaluated by harvesting and counting cells at preset time points after treatment with specific concentrations of caffeine.Flow cytometry was performed to assess cell cycle dynamics and apoptosis.Western blot analysis was used to detect the activity of the above signalling pathway.Results Caffeine treatment significantly suppressed GC cell growth and viability and induced apoptosis by enhancing the Caspase-9/3 pathway.Conclusion Caffeine can effectively promote the apoptosis of gastric cancer cells.