Aim To explore the effect of pemetrexed on autophagy and the function of autohpagy activation in A549 cells.Methods The anti-proliferative effect of pemetrexed on A549 cells was detected by MTS. The formation of autophagosome was observed by trans-mission electron microscopy (TEM)and the expression of autophagy-related proteins was determined by West-ern blot.The inhibitory effects of the combination of pemetrexed and a selective autophagy inhibitor on A549 cells were measured by MTS.Results Peme-trexed inhibited the proliferation of A549 cells.Auto-phagy was activated in A549 cells by exposure to pemtrexed.Autophagosome was increased in peme-trexed treatment cells compared with control group. Pemtrexed treatment led to the formation of autophago-some and the conversion of LC3-Ⅰto LC3-Ⅱ.The in-crease of Beclin-1 and the decline of autophagy sub-strate of p62 were observed in pemtrexed treated A549 cells.The pharmacological autophagy inhibitor chloro-quine (CQ)sensitized A549 cells to pemetrexed treat-ment.The inhibitory rate was (28.42 ±2.45 )% for pemetrexed only,and (45 .36 ±3.52 )% for peme-trexed combined with CQ.Conclusions Pemetrexed could activate autophagy in A549 cell lines.The dis-ruption of autophagy facilitates the anti-proliferative effect of pemetrexed on A549 cells,which provides a novel strategy in lung cancer treatment.

Objective To analyze the relationship between T lymphocyte subsets,neutrophil to lymphocyte ratio(NLR) and lung cancer pathological classification,clinical stage of non-small cell lung cancer.Methods The proportion of T lymphocyte subsets in 60 patients with lung cancer were detected by flow cytometry,the NLR were measured by blood sampling.Results The proportion of CD4+T lymphocytes,the proportion of CD8+T lymphocytes,CD4+/CD8+ and NLR in patients with lung adenocarcinoma,lung squamous cell carcinoma,small cell lung cancer patients were not statistically significant.The proportion of CD4+T lymphocytes and CD4+/CD8+ in patients with advanced non-small cell lung cancer(stage Ⅲ~Ⅳ) were lower(P<0.05) than in the early group(stage Ⅰ~Ⅱ).The proportion of CD8+T lymphocytes and NLR in patients with advanced non-small cell lung cancer(stage Ⅲ~Ⅳ) were higher(P<0.05) than that of early stage group(stage Ⅰ~Ⅱ).Conclusion The T lymphocyte subsets and NLR in the peripheral blood of non-small cell lung cancer patients have a certain correlation with the clinical stage of non-small cell lung cancer,which can be used as a reference index for clinical staging.

Objective To study how to reduce the complication rate of laparoscopic cholecystectomy (LC) for acute cholecystitis. Methods A retrospective analysis was made on clinical data of 158 cases of acute cholecystitis treated by LC from September 2001 to December 2003. Results Operations were accomplished laparoscopically in 151 cases, while conversions to open surgery were required in 7 cases (1 case of Mirizzi’s syndrome, 2 cases of gallbladder carcinoma, 1 case of cholecystoduodenal fistula, 2 cases of "ice-like" adhesions in the Calot triangle, and 1 case of common bile duct stones). Open re-exploration was performed in 1 case because of biliary leakage. Intraoperative cholangiography (IOC) was successfully conducted in 10 cases, 3 of which were found common bile duct stones. Out of the 3 cases, 2 cases underwent an intraoperative endoscopic sphincterotomy and 1 received a conversion to open choledochotomy with T-tube drainage. Conclusions When utilizing LC for acute cholecystitis, the rate of conversion and the incidence of complication may be reduced as long as the patients were properly selected, the principles of safety were abided by, and the intraoperative cholangiography was performed routinely.

Objective:To study the clinical value of left ventricle Tei index in evaluating hemodynamics after interventional therapy of patent ductus arteriosus (PDA).Methods:From May 2017 to May 2019, 50 children with PDA who underwent interventional therapy (PDA group) and 27 healthy children (healthy control group) in Anhui Provincial Children’s Hospital were selected. The left ventricle Tei index, plasma brain natriuretic peptides (BNP), left ventricular end-diastolic dimension (LVDD), left ventricular ejection fraction (LVEF) were compared between 2 groups.Results:The left ventricle Tei index was not correlated with heart rate and age in 2 groups ( P>0.05). The left ventricle Tei index before operation in PDA group was significantly lower than that in healthy control group: 0.20(0.16, 0.25) vs. 0.27(0.20, 0.30), and there was statistical difference ( P<0.05). In PDA group, the left ventricle Tei index immediately, 3 d, 1 month and 3 months after operation was significantly higher than before operation: 0.38(0.29, 0.47), 0.32(0.26, 0.40), 0.30(0.27, 0.35) and 0.32(0.26, 0.37) vs. 0.20(0.16, 0.25), and there was statistical difference ( P<0.05); the plasma BNP immediately after operation was significantly lower than before operation: 288 (126, 433) ng/L vs. 582 (303, 1 675) ng/L, and there was statistical difference ( P<0.05); the LVDD 3 months after operation was significantly lower than before operation: (3.03 ± 0.54) cm vs. (3.38 ± 0.51) cm, and there was statistical difference ( P<0.05); the LVEF immediately after operation was significantly lower than before operation: (54.24 ± 6.09)% vs. (59.45 ± 5.93)%, the LVEF 1 and 3 months after operation was significantly higher than that immediately after operation: (63.18 ± 4.71)% and (65.46 ± 4.78)% vs. (54.24 ± 6.09)%, and there were statistical differences ( P<0.05). The left ventricle Tei index before operation was negatively correlated with inner diameter of PDA and plasma BNP ( r = -0.362 and -0.388, P = 0.013 and 0.009), and there was no correlation between LVDD and LVEF ( r = -0.192 and -0.283, P = 0.229 and 0.053); the differences of Tei index before operation and immediately after operation (ΔTei) was positively correlated with inner diameter of PDA ( r = 0.325, P = 0.030), and there was no correlation with BNP, LVDD and LVEF ( r = 0.234, 0.283 and -0.039, P = 0.126, 0.076 and 0.798). Conclusions:The left ventricle Tei index can quickly and accurately assess the change of hemodynamics after interventional therapy of PDA.

OBJECTIVETo observe the effects of adipose-derived mesenchymal stem cells (ADSCs) with continous over-expression of glial cell line-derived neurotrophic factor (GDNF) on the motor function recovery and nerve regeneration of sciatic nerve of rats after electrical injury.

METHODSFive SD rats were collected to prepare ADSCs with over-expression of GDNF. One hundred and fifty SD rats were divided into normal control group (N), GDNF-ADSCs group (GA), ADSCs group (A), GDNF group (G), and physiological saline group (P) according to the random number table, with 30 rats in each group. Rats in group N were routinely fed without treatment, and rats in the other 4 groups were inflicted with electrical injury on sciatic nerve of thigh of the right hind leg. Rats in groups GA, A, G, and P were respectively injected with 100 µL suspension of ADSCs with over-expression of GDNF (1 x 10(7) cells per mL), 100 [µL ADSCs suspension (1 x 10(7) cells per mL), 100 µL GDNF solution (100 mg/L) , and 100 µL physiological saline to the surface of the injured nerves immediately after injury. Six rats of each group were collected for measuring hind limb stride from post injury week (PIW) 1 to 8, and morphology of the sciatic nerves was observed in PIW 8. In PIW 4, the protein expression of GDNF of sciatic nerves of the rest rats in each group was determined with Western blotting. Data were processed with one-way analysis of variance, analysis of variance of repeated measurement, and SNK test.

RESULTSCompared with that of group N, the hind limb stride values in groups GA, A, G, and P were significantly lower at each time point (with P values below 0.05). Compared with those of group P, the hind limb stride values in group GA from PIW 3 to 8, in group A in PIW 3, 5, and 7, and in group G in PIW 3, 5, 7, and 8 were significantly longer (with P values below 0.05). The hind limb stride values in group GA from PIW 4 to 8 were respectively (10.83 ± 0.97), (13.25 ± 1.40), (12.86 ± 1.42), (14.06 ± 1.50), and (15.09 ± 1.17) cm, which were significantly longer than those in group A [(8.90 ± 0.82), (9.03 ± 0.57), (9.27 ± 0.36), (9.86 ± 0.36), and (9.52 ± 0.58) cm] and group G [(8.87 ± 0.69), (8.51 ± 1.18), (9.34 ± 0.87), (9.76 ± 0.67), and (9.50 ± 1.22) cm], with P values below 0.05. Compared with that of group N, the number of myelinated nerve fibers of sciatic nerves was obviously decreased in group P but obviously increased in groups GA, A, and G; the diameter of axons was obviously shorter, and the myelin thickness was obviously increased in groups GA, A, G, and P in PIW 8 (with P values below 0.05). The number of myelinated nerve fibers in group GA was 31.2 ± 0.8, which was significantly higher than that in group A (23.7 ± 2.7), group G (22.3 ± 2.7), or group P (9.3 ± 2.8), with P values below 0.05. The diameter values of axons among groups P, A, G, and GA were similar (with P values above 0.05). The myelin thickness of rats in group GA was (3.41 ± 0.34) µm, which was significantly thicker than that in group A [(2.64 ± 0.37) µm] or group G [(2.41 ± 0.34) µm], with P values below 0.05. In PIW 4, the protein expression of GDNF of sciatic nerves was significantly higher in groups P, A, G, and GA than in group N (with P values below 0.05), and the protein expression of GDNF in group GA was significantly higher than that in group P, A, or G (with P values below 0.05).

CONCLUSIONSADSCs over-expressing GDNF protein can obviously promote the motor function recovery and nerve regeneration of sciatic nerve of rats after electrical injury.

Adipose Tissue ; Animals ; Electrophysiology ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; metabolism ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; metabolism ; Nerve Crush ; Nerve Regeneration ; physiology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; pathology ; physiology

Exosomes are generated by endosomal pathway when the multivesicular bodies fuse with the plasma membrane.Exosomes modulate gene regulatory network by transforming biological proteins,mRNA or miRNA to receptor cells.The research progress of stem cell-derived exosomes in the application of tissue repair and regeneration is reviewed in this paper.

BACKGROUND: Mesenchymal stem cells (MSCs), with low immunogenicity, can regulate cellular immunity and mitigate graft rejection, which has a good prospect in tissue engineering. However, it is rarely present in bone marrow. OBJECTIVE: To explore an isolation and culture method of the rabbit bone marrow-derived MSCs, to observe the biological characteristics and differentiation potential of bone marrow-derived MSCs.METHODS: MSCs were isolated from rabbit tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferation in vitro. Morphology was examined by phase contrast microscopy, and the growth curve of cultured MSCs was drawn via MTT results. MSCs were treated with osteogenetic inductor (L-DMEM/F12, 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 200 μmol/L vitamin C, 10 mmol/L β-phosphoglycerol), adipose inductor (L-DMEM/F12, 10% FBS, 1 μmol/L dexamethasone, 200 μmol/L antifani, 0.5 mmol/L IBMX, 10 μg/mL insulin), and chondrocytes inductor (L-DMEM/F12, 10% FBS, 10 μg/L TGF-β1, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C, 6.25 mg/L insulin) to differentiated into osteoblast, dipocytes and chondrocytes. And the differentiated cells were identified by alkaline phosphatase staining, oil red O staining, and toluidine blue staining, respectively.RESULTS AND CONCLUSION: Bone marrow-derived MSCs can be isolated and cultured by the combination of gradient centrifugation and different adherent method in vitro, which have the better potentiality of proliferation and multi-directional differentiation. Mostly of the primary and passaged cells were spindle-shaped. After osteogenetic induction, cells were positive to alkaline phosphatase staining. Oil red O staining showed that red lipid droplet existed in adipose cells, and toluidine blue staining showed that toluidine blue was positive after chondrocytes induction.

Objective To summarize the operational experience and explore the best surgery strategy in cicatricial alopecia.Methods Clinical date of 80 cases of cicatricial alopecia during January 2012 to December 2016 were analyzed retrospectively.The operation methods and related adverse events were recorded.Patients were followed-up on the postoperative 1 week,1 month,3 months,6 months and 1-2 years.The outcomes were evaluated by a 4-levels questionnaire:very satisfied,satisfied,not satisfied,and no effect.Results Forty cases were operated with expanded skin flap + Follicular unit extraction (FUE) transplantation,10 cases with scar resection + FUE repair,and 30 cases only with FUE.Twenty cases were completed treatment with single-stage surgical operation,and 60 cases with two-stage surgical operation.A percentage (70％) of patients was very satisfied and 30％ were satisfied after one-stage surgical operation.A percentage (85％) of patients was very satisfied and 15％ were satisfied after two-stage surgical operation.Conclusions The cicatricial alopecia needs comprehensive surgical treatment.FUE is a best additional operation technology.The effect of combined treatment is better than single therapy method in large area cicatricial alopecia.

Hirsutine,an indole alkaloid extracted from the traditional Chinese herb Uncaria,has exhibits diverse pharmaco-logical activities,including antihypotensive,antiinfective,and cardioprotection.In cervical cancer,breast cancer,colon cancer,and oth-er malignant tumors,hirsutine has effects on anti-tumor by regulating cell apoptosis,immune regulation and other pathways,laying the foundation for the application of hirsutine in the field of cancer therapy.This article reviews relevant literature both domestically and internationally to elucidate the research progress of hirsutine in anti-tumor therapy,providing new ideas and strategies for cancer treat-ment.

OBJECTIVE:To provide reference for rational use of tigecycline.METHODS:Based on tigecycline instructions,referring to related specifications and literatures,through pharmacy and clinical expert discussion,DUE criteria for tigecycline was established from medication indications,medication process,medication results and administrative supervision.In retrospective study,DUE criteria was used to evaluate medical records of inpatients in our hospital during Jan.2015-Dec.2016.RESULTS:A total of 71 medical records of inpatients receiving tigecycline were included.The use of tigecycline in our hospital was basically up to DUE criteria.But there still was unreasonable phenomenon,such as microbial inspection rate was 81.7％ (aiming at 90％);the rate of medication course meeting the criteria was 76.1％ (aiming at 90％);the proportion of patients with consultation records was 81.7％ (aiming at 100％);the rate of prescribing authority meeting the criteria was 85.9％ (aiming at 100％).CONCLUSIONS:The established DUE criteria for tigecycline shows strong operability and practicability,and provide reference for the work development of doctors and clinical pharmacists.