Objective To investigate the status of risk population of diabetic foot in community and analyze the influencing factors.Methods 210 diabetes patients living in a community in Shanghai were enrolled,acoording to Gavin's weighted integral method with risk factors of diabetic foot,Doppler blood flow detector,Semmes Weistein 5.07 (10 g) nylon monofilament and 128Hz tuning fork were used to screen for the risk population of diabetic foot.Results Risk population of diabetic foot were 174 patients (82.9％),wherein the low risk group were 112 patients (53.4％),in the middle and high risk group were 62 people (29.5％).In addition to the Gavin's diabetic foot risk factors,different ages,cultural degrees,the values of FBG,PBG and HbAlc would also affect the risk level of diabetic foot.While the values of FBG,PBG and HbAlc of diabetes patients in the moderate and high risk group were significantly higher than those in the normal group.Conclusions We should make early screening for risk factors of diabetic foot and strengthen health education and management in order to effectively prevent diabetic foot.

ObjectiveTo determine whether glutamate induces the alteration of aquaporin 4 (AQP4) expression in cultured rat astrocytes. MethodsThe secondary cultured astrocytes were treated with 1 mmol/L L-glutamate for 1 h, 3 h, 6 h, 12 h, 24 h and 48 h. The morphologic changes of astrocytes were observed through microscope after GFAP immunostaining and AQP4 mRNA expression were detected with real-time PCR. ResultsThe astrocytes swelled when exposed to glutamate for 1 h and remained with prolonged treatment. Meanwhile, the AQP4 mRNA expression were early down-regulated and subsequently up-regulated, featured with the lowest AQP4 mRNA level at 12 h after treatment (P<0.01) and higher at 48 h (P<0.05). ConclusionAquaporin 4 may be involved in the occurrence and development of astrocyte swelling induced by glutamate.

Objective To compare the differences between the cell swelling of cultured astrocytes ( AST) from Wistar and Sprague-Dawley ( SD) rats after incubation with glutamate.Methods Primary cultured AST derived from the cerebral cortex of one-day-old Wistar or SD rats were prepared.The cultured AST received 1 or 10 mmol/L glutamate treat-ment for 48 h on the tenth day after subculture.The viability of AST was determined by lactate dehydrogenase ( LDH) kit to assess the cell injury, and the perimeter of AST was measured using Image Pro Plus software after glial fibrillary acidic protein immunofluorescence staining to evaluate the astrocyte swelling.Then, the expression of aquaporin 4 ( AQP4 ) in cultured AST was detected by quantitative reverse transcription polymerase chain reaction.Results No significant differ-ence was found in the LDH release after the glutamate treatment in cultured AST from these two strains (P>0.05).The perimeter of AST from normal Wistar rats was shorter than that from SD rats, but was longer after the treatment of glutamate (P<0.05).Meanwhile, AQP4 expression in the Wistar rats was significantly higher than that from SD rats after incuba-tion with 1 mmol/L glutamate ( P<0.05 ) .Conclusions These results suggeste that cultured AST from Wistar rats are more susceptible to glutamate-induced swelling than that from SD rats, and there are differences between the effects of glu-tamate on AQP4 expression in astrocytes of Wistar and SD rats.

Objective To explore the expression of aquaporin 4 (AQP4) in primary and secondary cultured rat astrocytes. Methods Rat cortical astrocytes from a newborn (one day) Wistar rat were cultured. Astrocytes were identified with immunofluorescence staining of glial fibrilillary acidic protein (GFAP). The expression of AQP4 was determined with real-time quantitative polymerase chain reaction and immu-nofluorescence staining three, five, seven and nine days of primary culture, and nine days of secondary culture. Results The purity of GFAP-positive cells was more than 95％. The expression of AQP4 mRNA was found three days of primary culture, remained unchanged five days of primary culture (P>0.05), and increased seven and nine days of primary culture (P<0.05). The expression of AQP4 mRNA was not different between nine days of primary culture and nine days of secondary culture (P>0.05). AQP4 immunofluorescence staining showed the same trend of AQP4 mRNA. Conclusion AQP4 may express since three days of primary culture in rat astrocytes in vitro, and increase slowly until nine days of primary culture.

Objective To investigate the protective effect of methylene blue (MB) on blood-brain barrier (BBB) injury after focal cere-bral ischemia-reperfusion in rats. Methods 18 male Sprague-Dawley rats were randomly divided into sham-operated group (n=6), model group (n=6) and MB treatment group (n=6). The left middle cerebral arteries were occluded for 1 hour and reperfused. MB was infused intra-venously immediately after reperfusion (3 mg/kg) and again 2 hours post-reperfusion (1.5 mg/kg), while normal saline was administered in the model group. The sham-operated group was treated as same as the model group without occlusion and infusion. HE staining was used to observe the histological injury in the cortex around the infarcted region 47 hours after reperfusion, while albumin immunohistochemistry was used to evaluate the permeability of the BBB, and immunohistochemistry and double immunofluorescence staining were used to exam-ine the expressions of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4). Results HE staining showed that cells and blood ves-sels were not intact in the cortex around the infarcted region in the model group and they were better in the MB treatment group. The expres-sions of the albumin, GFAP and AQP-4 were higher in the model group than in the sham-operated group (P<0.01), and were lower in MB treatment group than in the model group (P<0.05). The double immunofluorescence staining showed the colocalization of GFAP and AQP-4 in the astrocytes. Conclusion MB may ameliorate the BBB disruption induced by focal cerebral ischemia-reperfusion through reducing glio-cyte proliferation and down-regulation of AQP-4 expression in rats.

Objective To examine the sensitivity of human glioblastoma multiforme cell line BT325 to 5 antineoplastic drugs, including cisplatin (DDP), teniposide (VM26), nimustine (ACNU), temozolomide (TMZ) and vincristine (VCR). Methods BT325 cells were incubated in DMEM with 10% or 20% fetal bovine serum (FBS) or without FBS respectively. The cell numbers were counted at 24 h, 48 h, 72 h,96 h, 120 h, and 144 h, then platting and growth curve were drafted. Cell counting kite-8 was used to detect the influence of 5 drugs with different concentrations on human glioma cell line BT325. Results DDP and VM26 significantly suppressed BT325 cells(>75%) viability in a dose-dependent manner, while VCR inhibited BT325 cells (50%) growth without dose-effect relationship. In contrast, ACNU and TMZwere not effective on the viability of BT325 cells. Conclusion BT325 cells were very sensitive to chemotherapeutic drugs DDP amd VM26.

Objective To investigate the expression changes of astrocytic syntrophin in hippocampus from human mesial temporal lobe epilepsy (MTLE). Methods From April, 2015 to July, 2016, 17 cases of hippocampus, collected from temporal lobectomy, were divided into MTLE group (n=13) and non-MTLE group (n=4) according to hematoxylin and eosin staining, glial fibrillary acidic protein and neuronal nu-clei immunohistochemical staining. Immunofluorescence double labeling and immunofluorescence histochemistry were used to observe the expression of syntrophin. Results The proliferation of astrocytes increased and neurons reduced in the hippocampus of MTLE group. Syntro-phin was found in the membrane and foot processes of astrocyte, that was enriched along perivascular astrocyte end-feet domain in non-MTLE group, but lost in MTLE group. While the whole expression of syntrophin was more in MTLE group than in non-MTLE group (t=5.421, P<0.001). Conclusion The distribution of syntrophin in hippocampus astrocytes may be related to the development of MTLE.

AIM: To study the distribution of CYP2C9∗3 and VKORC1-1639G>A gene polymorphism in Han population in Anhui province and their influence on the stable dose of warfarin. METHODS: The blood samples of 1 169 patients from 6 tertiary general hospitals in 5 areas of Anhui province from January 2020 to December 2021 were selected, the genotype of CYP2C9∗3 and VKORC1-1639G>A was detected by fluorescent staining in situ hybridization technique. RESULTS: The distribution of CYP2C9∗3 genotypes in 1 169 patients: the frequencies of AA, AC and CC genes were 90.16%, 9.24% and 0.60%, respectively; The distribution of VKORC1 genotype: the frequencies of AA, AG and GG genes were 84.26%, 14.71% and 1.03% respectively; There was no significant difference between the two genotypes in gender, age and regional distribution (P>0.05). The average daily warfarin dose of CYP2C9∗3 AA genotype in 755 patients with stable warfarin dose was (3.02±0.59) mg/d, which was significantly higher than patients with AC genotype and CC genotype; The average daily warfarin dose of patients with VKORC1-1639AA genotype was (2.72±0.40) mg/d, which was significantly lower than that of patients with AG genotype and GG genotype (P<0.05). And the difference was statistically significant (P<0.05); There are significant differences in gender, age and clinical diagnosis between patients with stable dose of warfarin and those without stable dose (P<0.05). CONCLUSION: CYP2C9 and VKORC1 genotypes are associated with the stable dose of warfarin. Clinical anticoagulation therapy guided by CYP2C9 and VKORC1 genotypes can provide guidance for individualized medication of warfarin.