Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

Purpose: Hungry bone syndrome after parathyroidectomy is an important clinical problem in patients on maintenance hemodialysis. We examined the effect of an enhanced recovery after surgery (ERAS) program on the incidence of hungry bone syndrome after parathyroidectomy in this population. Methods: This single-institution, retrospective study analyzed 108 patients on hemodialysis who underwent parathyroidectomy for secondary hyperparathyroidism. Patients were classified into the pre-ERAS (n = 52) and post-ERAS (n = 56) groups. The ERAS program identified high-risk patients and enforced aggressive measures to normalize calcium levels following parathyroidectomy. Results: There was no significant difference in age, sex, body weight, presenting symptoms, preoperative calcium and alkaline phosphatase levels, postoperative intact parathyroid levels, postoperative calcium levels at 1 and 24 hours after parathyroidectomy, and 30-day readmission rates between the groups. The post-ERAS group had significantly higher levels of postoperative calcium at 48 and 72 hours after parathyroidectomy, but a lower incidence of hungry bone syndrome and shorter postoperative length of stay. Patients with hungry bone syndrome had higher preoperative levels of alkaline phosphatase and intact parathyroid, longer postoperative length of stay, and were less likely to have been part of the ERAS program. High preoperative alkaline phosphatase levels and absence of the ERAS program were independent risk factors for hungry bone syndrome after parathyroidectomy. Conclusion The ERAS program reduced the incidence of hungry bone syndrome and shortened the postoperative length of stay in patients on maintenance hemodialysis who underwent parathyroidectomy for secondary hyperparathyroidism.

【Objective】 To observe the effects of hsa-miR-124-3p.1 in inhibiting epithelial-mesenchymal transition （EMT）， migration and invasion of human gastric cancer cells induced by transforming growth factor β1 （TGF-β1） by targeting tumor necrosis factor receptor-associated factor 6 （TRAF6）. 【Methods】 A total of 43 gastric cancer tissues and 43 normal para-carcinoma tissues were collected. The human gastric mucosal epithelial cells GES-1 and gastric cancer cells （NCI-N87， MGC-803， BGC-823， SGC-7901， and MKN-45） were cultured. The expressions of miR-124-3p.1 and TRAF6 in tissues and cells were detected by fluorescent quantitative PCR and Western blotting. The targeted relationship between miR-124-3p.1 and TRAF6 was verified by dual-luciferase reporter gene system assay. SGC-7901 cell lines with miR-124-3p.1 and TRAF6 overexpression were constructed. The cells were induced by TGF-β1. The invasion and migration abilities of the cells were evaluated by Transwell chamber assay and scratch test. 【Results】 Compared with normal para-carcinoma tissues and normal gastric mucosal cells， the expression of miR-124-3p.1 was downregulated， while the expressions of TRAF6 mRNA and protein were upregulated in gastric cancer tissues and cells （P<0.05）. Compared with control group， expression of E-cadherin in cells was downregulated， expressions of N-cadherin and Vimentin were upregulated， invasion and migration rates of cells were increased in TGF-β1 group （P<0.05）. Compared with TGF-β1 group， after cells were transfected with miR-124-3p.1 mimic， the expression of E-cadherin was upregulated， the expressions of N-cadherin and Vimentin were down-regulated， and invasion and migration rates of cells were decreased （P<0.05）. Compared with miR-124-3p.1 mimic group， invasion and migration rates of cells were increased in TGF-β1+mimic+TRAF6 group， expressions of TRAF6， N-cadherin and Vimentin were up-regulated， and the expression of E-cadherin was down-regulated （P<0.05）. 【Conclusion】 hsa-miR-124-3p.1 is lowly expressed in gastric cancer. Overexpression of miR-124-33p.1 can inhibit EMT， cell invasion and migration induced by TGF-β1. And the action mechanism may be related to the downregulated expression of TRAF6.

Objective To investigate the application of MSCT three-dimensional digital navigated biopsy in subcarinal lesions.Methods 82 patients were enrolled.Study subjects were randomly divided into control group and research group.Three-dimensional positioning and three-dimensional navigation needle biopsy were used in research groups, while CT cross-sectional image positioning with conventional puncture needle was used in control group.Puncture accuracy, one-time success rate of puncture, complications, diagnosis accuracy and operation time were compared between the two groups.Results Puncture success rate, definite diagnosis rate were 87.80%(36/41) and 97.56%(40/41) for the research group,and 60.97%(24/41) and 80.49% (33/41) for the control group, respectively,which on the research group were higher than that on the control group(χ2=8.945, 6.116;P<0.05).Complication rate and operating time were 14.63% (6/41) and (11.64±2.76) min for the research group, and 41.45% (17/41) and (22.22±6.31) min for the control group, respectively, which were lower on the research group than that on the control group (χ2=7.31,t=-11.70,P<0.05).Conclusion MSCT three-dimensional digital navigated biopsy technique could promote the efficiency of subcarinal space puncture biopsy significantly,which is a novel, convenient, precise and safe method.

OBJECTIVE:To provide reference for rational drug use in the clinic. METHODS:Irrational medication orders eval-uated by the Affiliated Cancer Hospital of Zhengzhou University during Oct. 2014 to Sep. 2015 were arrangemented,summarized and analyzed. RESULTS:A total of 515 inpatient medical records were reviewed and analyzed,among which there were 165 unrea-sonable medical records and 185 irrational medication orders. Irrational medical records of general surgery department were the most(38 items,accounting for 23.03%). Irrational drug use mainly included irrational usage and dosage(80.00%),drug use with-out indications or not suit indications (7.57%),inappropriate solvent selection (4.86%). Including 66.22% of single overdose, 18.92% of longer medication duration. CONCLUSIONS:There are many irrational medical orders which should be standardized in our hospital,especially overdose and longer medication duration,which increase financial burden of patient. Pharmacists should strengthen communication with clinicians,and hold rational drug use trainings regularly base on the types of the irrationality. These can help to improve rational drug use and guarantee the safety of drug use.

Objective To study the effect of clinical pharmacist interventions on the rational use of antibiot-ics.Methods 1 000 hospitalized patients before the implementation of intervention in April 2012 to April 2013 (control group)and the other 1 000 cases of hospitalized patients in May 2013 to May 2014 after the implementation of the intervention (study group)were selected,the antibiotics use rate,hospitalization time and cost,cost of using antibacterial drugs were compared between the two groups.Results The antibiotics use rate of the study group was 56.0%(560/1 000),significantly lower than 78%(780/1 000)of the control group,with significant differences between the two groups(χ2 =11.089,P=0.032),the hospitalization time,cost of using antibacterial drugs of the study group were (12.6 ±0.8)days,(912.2 ±13.2)yuan,significantly better than (16.9 ±0.7)days,(1 528.1 ± 32.5)yuan of the control group,with significant differences between the two groups (t=9.892,10.142,P=0.028, 0.014);The two groups of hospitalization expenses showed no statistical significance (t =4.984,P =0.072 ). Conclusion Clinical pharmacist intervention has a positive effect on the application of antibacterial drugs,which can significantly reduce the use of antimicrobial drugs and reduce the hospitalization days.

Objective To investigate the correlation between asymmetric dimethylarginine (AD-MA) and endothelial dysfunction in patients with uremia.Methods Uremic patients who did not receive hemodialysis were defined as A group (n =40) ; uremic patients who had received hemodialysis were divided into B group (n =45) ;healthy people were defined as C group (n =20) ;and chronic kidney disease (stage 2 ～ 4) patients were defined as D group (n =20).The diameter of intima-media thickness,and endothelium-dependent or independent dilation (EDD or EID) of radial artery in right forearm were detected with diasonography.The levels of ADMA were determined by high-performance liquid chromatography.Results Compared to C group,the levels of ADMA in A,B and D groups were significantly increased [C:(0.78 ±0.19) μmol/L,A:(1.51 ±0.16) μ mol/L,B:(1.13 ±0.14) μmol/L,D:(0.92 ±0.11) μmol/L; P ＜0.05].Compared to A group,the levels of ADMA were significantly decreased in B group (P ＜0.05).EDD and EID were decreased significantly in A,B and D groups compared to C group [EDD:C:(13.52±1.73)％ vs A:(7.32 ±0.54)％,B:(9.02 ±0.86)％,D:(10.13 ±1.25)％,P ＜0.05;EID:C:(14.45±1.85)％ vsA:(10.37 ±1.51)％,B:(9.54±1.39)％,D:(11.17±1.56)％,P ＜0.05].EDD in B group was significantly lower than A group (P ＜0.05).In group A,a negative correlation was found between EDD and the level of ADMA (r =-0.81,P =0.020).Conclusions ADMA level was significantly increased in uremic patients.A close correlation existed between ADMA and endothelial dysfunction of radial artery.

Objective To investigate the role of surfactant protein (SP)-A and SP-D in urinary tract infection mouse model,and evaluate the effects of SP-A and SP-D absence on urinary tract infection.Methods SP-A and SP-D double knockout (SP-A/D KO) mice were made.SP-A/D KO and wild-type (WT) C57BL/6 female mice were used for this study.The expression of SP-A and SP-D in kidney was detected by immunohistochemistry (IHC).The levels of p-p38 and p38 protein in kidneys were measured by Western blotting.Uropathogenic Escherichia coli or buffer was delivered into the bladder of female mice.At 24 and 48 h after inoculation,CFU of Escherichia coli in the kidney and urine of the treated and control mice were measured.Histological,cellular and molecular analysis were performed by several methods of H/E staining,IHC and Western blotting.The effects of SP-A and SP-D on bacterial growth were studied in vitro.Results SP-A and SP-D in kidney were located in the proximal tubules and collecting tubules.Compared with WT mice,infected SP-A/D KO mice with UPEC had higher CFU in kidneys and urine at 24 h and 48 h,increased inflammatory cells infiltration in kidneys (P＜0.05).Compared with WT mice,SP-A/D KO mice had higher p38 MAPK phosphorylation levels in kidneys (P ＜ 0.05).Growth of Escherichia coli was greatly inhibited by both SP-A and SP-D (P＜0.05).Conclusions Both SP-A and SP-D are expressed in kidney.SP-A and SP-D can attenuate UTI induced by UPEC which may be through inhibiting bacterial growth and modulating renal inflammation.

Objective To investigate the effect of angiotensin 1-7(Ang 1-7) on renal proximal tubular epithelial cell(HK-2) transdifferentiation induced by high glucose.Methods All the raised HK-2 cells were divided into 5 groups: normal control group,high glucose group,high glucose with Ang1-7 group,high glucose with Ang1-7 and A779 group,high glucose with pioglitazone group.Expression of peroxisome proliferator activated receptor-γ(PPAR-γ) and α-smooth muscle actin(α-SMA) was detected by Western blotting,real-time PCR and immunofluorescence.Results The levels of PPAR-γ protein and mRNA in HK-2 cells were significantly increased after treatment with high glucose and Ang 1-7.Expression of α-SMA protein and mRNA was inhibited remarkably after treatment with high glucose and Ang 1-7.These effects of Ang 1-7 on HK-2 cells could be reversed by Mas receptor antagonist A779.Conclusion Ang 1-7 inhibits high glucose-induced expression of o-SMA in HK-2 cells,which is in part through the Mas.