Objective To investigate the effect of microRNA-204(miR-204)on autophagy in U251. Methods Inhibition of miR-204 in U251 cell lines was done by transfection of miR-204 inhibitor(AMO-204) .Cell viability was detected by MTT assay.The autophagy of U251 was tested by immunofluorescence tech-nique.The protein level of Beclin 1,LC3 and Bcl-2 was detected by Western blot.Results Cell viability was markedly increased after inhibition of miR-204 in U251 cells.The number of autophagosome was decreased.The levels of Beclin 1 and LC3 were decreased,the protein level of Bcl-2 was significant increased by transfection of AMO-204 in U251 cells(P<0.05).Conclusion MiR-204 might at least in part promote glioma via inhibi-ton autophagy,indicating that miR-204 might be a potential target for the treatment of glioma.

BACKGROUND: The blocking treatment can improve the clinic symptom of facial spasm. But the site, depth and the angle of puncturing point are very difficult to be defined, which will seriously infect the curative effect.OBJECTIVE: To study the applied anatomy of the related structures of facial muscle for blocking the facial nerve, and to provide anatomical bases for accurate puncturing point and preventing complications.DESIGN: An observation study based on cadavers and making the mimic puncture and measuring the correlative structures with anatomical method.SETTING: Department of anatomy in a medical college.PARTICIPANTS: Totally 28 adult male cadavers(56 sides) were used and the correlative index were measured.METHODS: The blocking point was located at the convergent point of the interior edge of cartilage of the external acoustic canal, the anterior fringe of mastoid processes and the posterior fringe of mandible ramus, the needle must be thrust vertically to the median sagittal plane. The puncturing needle stopped until it was barred. A longitudinal incision was made from the puncturing point to mandibula angle, and skin, superficial fascia were cut separately. Then carotid superficial fascia was cut from the posterior fringe of parotidean. The facial nerve trunk and the posterior auricular artery were preserved in site. A blunt isolation was made along its incision. Jugular glomus was appeared. Then the correlative index were measured and dealt with statistics method.MAIN OUTCOME MEASURES: The puncturing point, angle and depth,the distance between facial nerve trunk and puncturing point, the position relationship between facial nerve and puncturing needle and the distance relationship between facial nerve and principal structures adjacent to it.RESULTS: The blocking point was located at the convergent point of the interior edge of the external acoustic canal, the anterior fringe of mastoid processes and the posterior fringe of mandible ramus, the needle must be thrust vertically to the median sagittal plane. On the left side, the puncturing depth was (19.91 ± 0.09) mm, On the right side, the puncturing depth was(19.28±0. 10) mm.CONCLUSION: The experimental study could provide accurate puncturing point, angle and depth for blocking the facial nerve and bring the blocking treatment into full play.

Objective To study the expression of CD44 gene in non-small cell lung cancer(NSCLC) tissue and the relationship with prognosis.Methods The expressions of CD44 gene in 36 specimens from 36 patients with NSCLC were determined by RT-PCR and all the patients were followed up for 3 years.Results CD44 gene was excessively expressed in 21 of 36 specimens of NSCLC tissues,excessive expression rate was 58.3%.The expression of CD44 mRNA in NSCLC tissue was related to metastasis of lymph node(?2 =9.787,P

Objective To evaluate the therapeutic effect of hydrogen-rich saline on allergic contact dermatitis (ACD) in mice and to explore its underlying mechanisms.Methods Forty mice were equally divided into 4 groups:control group,control treatment group,ACD group and ACD treatment group.ACD was induced by repetitive topical application of dinitrofluorobene (DNFB) to the left ear of mice on day 1,2 and 5.Hydrogen-rich saline was intraperitoneally given to the mice in the ACD treatment group at a dose of 5 ml/kg per day from day 1 to 5.On day 6,the mice were sacrificed,ear tissue was removed from them and subjected to measurement of thickness and weight,detection of tumor necrosis factor (TNF)-α,interleukin (IL)-6,IL-17 and interferon (IFN)-γ expression by enzyme linked immunosorbent assay,as well as numeration of inflammatory cells in lesions after hematoxylin-eosin (HE) staining.Data were processed by SPSS 18.0 software,and statistical analysis was carried out by one-way analysis of variance and least significant difference (LSD) procedure.Results Compared with the control group,the ACD group showed a significant increase in lesion score (7.33 ± 1.53 vs.0,P ＜ 0.05),differences in the thickness ((0.73 ± 0.15) mm vs.(0.13 ± 0.05) mm,P ＜ 0.05) and swelling degree (expressed as tissue weight:(18.67 ± 3.05) mg vs.(3.33 ± 1.52) mg,P ＜ 0.05) between the left and right ear,expressions of TNF-α ((1475.52 ± 233.81) pg/mg vs.(239.01 ± 52.39) pg/mg,P＜ 0.05),IL-6 ((184.65 ± 78.39) pg/mg vs.(42.28 ± 17.64) pg/mg,P＜ 0.05),IL-17 ((628.56 ± 201.44) pg/mg vs.(127.58 ± 50.28) pg/mg,P＜ 0.05) and IFN-γ ((197.72 ± 37.81) pg/mg vs.(24.57 ± 8.31) pg/mg,P ＜ 0.05),and the number of inflammatory cells per square millimetre in the left ear tissue (752.00 ± 166.06 vs.127.33 ± 77.18,P ＜ 0.05).However,hydrogen-rich saline treatment induced a statistical decrease in all of these parameters in the ACD treatment group compared with the ACD group,including lesion score (3.33 ± 0.58,P ＜ 0.05),difference in thickness ((0.46 ± 0.11) mm,P ＜ 0.05) and swelling degree ((11.00 ± 2.64) mg,P ＜ 0.05),expressions of TNF-α ((817.72 ± 101.13) pg/mg,P＜ 0.05),IL-6 ((95.86 ± 36.65) pg/mg,P＜ 0.05),IL-17 ((373.38 ± 126.74) pg/mg,P＜ 0.05),IFN-γ ((63.31± 17.38) pg/mg,P ＜ 0.05) and the number of inflammatory cells per square millimetre (384.00 ± 97.35,P ＜0.05).Conclusion Hydrogen may inhibit the release of inflammatory factors in ACD.

Objective To establish the phenobarbital addiction rat model by the modified drug‐admixed‐food(DAF) method . Methods Rats were selected as the experimental animal .The modified DAF method was adopted to construct the model .The initial dose of phenobarbital 75 mg/kg was given mixed and the dase was gradually increased day by day ,which continued for 45 d .The es‐tablished model was verified by the precipitated withdrawal method and the natural withdrawal method for judging whether the model construction succeeding .Results The convulsion rate in the DAF precipitated withdrawal group was higher than that in the control group(P< 0 .01) ;the withdrawal symptoms scores in the DAF natural withdrawal group were higher than those in the con‐trol group(P< 0 .05) ;the relieving degree of body mass in the two withdrawal groups was greater than that in the control group (P< 0 .05) .The brain section in the mode group revealed obvious cellular apoptotic change .Conclusion DAF is simple and feasible for establishing the phenobarbital addiction rat model .

Objective To investigate whether hydrogen can regulate the expressions of inflammatory factors by ultraviolet B (UVB)-induced human HaCaT keratinocytes through the autophagy pathway.Methods Cultured HaCaT keratinocytes were divided into several groups:blank control group receiving no treatment,hydrogen group cultured in hydrogen-rich medium,three UVB groups irradiated with UVB at 1,10,50 mJ/cm2 respectively,three UVB + hydrogen groups irradiated with UVB at 1,10,50 mJ/cm2 respectively followed by culture in hydrogen-rich medium,UVB + 3MA group pretreated with the autophagy inhibitor 3MA for 1 hour followed by UVB radiation at 50 mJ/cm2,UVB + rapamycin group pretreated with the autophagy activator rapamycin for 1 hour followed by UVB radiation at 50 mJ/cm2,UVB + 3MA +hydrogen group pretreated with 3MA for 1 hour followed by UVB radiation at 50 mJ/cm2 and culture in hydrogen-rich medium,UVB + rapamycin + hydrogen group pretreated with rapamycin for 1 hour followed by UVB radiation at 50 mJ/cm2 and culture in hydrogen-rich medium.After additional culture with or without hydrogen for 12 hours,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cellular proliferative activity,Western-blot analysis to measure the expressions of autophagy-associated protein 1 light chain 3 (LC3) and Beclin 1,and enzyme-linked immunosorbent assay (ELISA) to measure the supernatant levels of inflammatory factors including tumor necrosis factor (TNF)-α,interleukin (IL)-1β,IL-6 and high mobility group protein B1 (HMGB1),and a test kit was used to determine the level of lactate dehydrogenase (LDH).Results Compared with the blank control group,the 10-and 50-mJ/cm2 UVB groups showed significantly increased release of LDH,expressions of LC3 and Beclin1 and supernatant levels of TNF-α,IL-1 β,IL-6 and HMGB 1,but decreased cellular proliferative activity (all P ＜ 0.05).Hydrogen significantly attenuated the release of LDH,down-regulated the supernatant levels of TNF-α,IL-1β,IL-6 and HMGB1,but up-regulated cellular proliferative activity as well as LC3 and Beclin1 expressions in the 10-and 50-mJ/cm2 UVB + hydrogen groups compared with the 10-and 50-mJ/cm2 UVB groups respectively (all P ＜ 0.05).In addition,the levels of TNF-α,IL-1β,II-6 and HMGB1 were significantly higher in the 50-mJ/cm2 UVB + 3MA group than in the 50-mJ/cm2 UVB group,and higher in the 50-mJ/cm2 UVB + 3MA + hydrogen group than in the 50-mJ/cm2 UVB + hydrogen group,but lower in the 50-mJ/cm2 UVB + rapamycin group than in the 50-mJ/cm2 UVB group (all P＜ 0.05).Conclusion UVB radiation can increase the expressions ofautophagy-associated proteins,and hydrogen-rich medium can down-regulate the expressions of inflammatory factors by UVB-induced HaCaT cells through the autophagy pathway.

Objective To compare the three different methods of enzyme linked immunosorbent assay(ELISA),to select the best method for clinical diagnosis and treatment.Methods Addcare ELISA800,TECAN freedom evolyzer and manual ELISA method were used to detect hepatitis B virus Pre S1 antigen(preS1Ag) hepatitis B virus Pre S2 antigen(preS2Ag) in confrontation control product samples and serum specimens from patients with HBV,and the results were analyzed by statistical methods.Results The batch precisions of the three methods to detect pre-S1Ag were 4.73%,5.38%,11.87%,the batch precisions of the three methods to detect pre-S2Ag were 4.91%,5.04%,11.75%.The inter batch precisions of the three methods to detect pre-S1Ag were 6.63%,7.90%,13.26%,the inter batch precisions of the three methods to detect pre-S2Ag were 6.74%,7.81%,12.59%.All the sensitivities were 100.00%.Conclusion All the three methods have good consistency,which could be used in the detection of Pre-S1Ag and Pre-S2Ag.The precision of Addcare ELISA800 is the best,which could further improve the quality of clinical testing.

Objective To observe the effect of hydrogen on ultraviolet B (UVB)-induced oxidative damage to skin fibroblasts.Methods Primary human skin fibroblasts from foreskin tissues were divided into five groups:normal control group receiving no treatment,hydrogen control group treated with hydrogen-rich saline,UVB group receiving irradiation only,post-treatment group irradiated with UVB followed by hydrogen-rich saline treatment,and pre-treatment group treated with hydrogen-rich saline followed by UVB irradiation.The dose of UVB was 30,60 and 90 mJ/cm2 in the cell proliferation assay and 90 mJ/cm2 in the other experiments.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of fibroblasts,a chemiluminescence method to estimate the activity of superoxide dismutase (SOD) and catalase as well as to determine the level of malondialdehyde in the culture supernatant of fibroblasts,enzyme linked immunosorbent assay (ELISA) to determine the supernatant level of 8-isoprostane-prostaglandin F2α (8-iso-PGF2α),Western blot to detect the expression of heme oxygenase-1 (HO-1) in fibroblasts.One-factor analysis of variance was conducted to assess differences in these parameters among these groups.Results UVB irradiation decreased the proliferative activity (absorbence value at 490 nm) of fibroblasts in a dose-dependent manner.Both the pre-treatment group and post-treatment group showed a statistical increase in proliferative activity of cells compared with the corresponding UVB control groups (all P ＜ 0.05).The activity of SOD and catalase as well as the protein expression of HO-1 were significantly higher (all P ＜ 0.05),whereas the supernatant levels of malondialdehyde and 8-iso-PGF2α were statistically lower (both P ＜ 0.05) in the pre-treatment group and post-treatment group than in the UVB control group.Conclusion Hydrogen may mitigate UVB-induced oxidative damage to skin fibroblasts.

Objective To evaluate the effect of operation on breast duct fishtula. Methods 41 patients with breast duct fitula were subjected to fistulectomy or mastectomy. Results All patients had no re ccurrence after operation from 0.5 to 17 years. The clinical analysis showed that the causes of breast duct fistula were bacterial infection, retracted nipple, tissuration in the middle of nipple and breast duct dialation. Conclusions Fistulectomy or mastectomy is the most effective treatment of breast duct fistula.

Objective To evaluate the effects of modified method of fixation and withdrawal of needles in intravenous infusion.Methods The modified method of two-step fixation and withdrawal of needles was adopted in emergency transfusion room of our hospital from September 2013.In January 2015,during every day's low peak period 12:00-14:00,246 emergency transfusion patients were chosen and divided into the traditional group (122 cases,using three-step fixation and withdrawal of needles) and the modified group (124 cases,using two-step fixation and withdrawal of needles) according to transfusion order.The pain degree caused by withdrawal of needles using Numeric Rating Scale (NRS),time consumption of fixation and withdrawal of needles and rate of adhesive pastes abscission was assessed by nurses and compared between two groups.Results The incidence of pain in the modified group was lower than that of the traditional group [48.39％ (60/124) vs.81.97％ (100/122)],x2=30.49,P＜0.05.The time consumption of fixation and withdrawal of needles in the modified group was shorter than that of the control group [(7.55 ±2.01) seconds vs.(10.88 ±2.72) seconds;(2.44 ±0.84) seconds vs.(11.55 ± 4.62) seconds],Z=8.70,13.55,P＜0.05.Therate of adhesive pastes abscission in the modified group was lower than that of the control group [4.0％(5/124) vs.18.9％ (23/122)],x2 =13.39,P＜0.05.All the difference between two groups was statistically significant.Conclusions The modified method of fixation and withdrawal of needles can relieve the pain caused by withdrawal of needles.Nurses can operate easily,adhesive pastes is fixed sturdily,which is popular among nurses and patients.