AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.

Objective To explore the clinical application of microsurgical techniques to therapy the femoral neck fracture in younger patients and prevent the necrosis of femoral head.Methods Seventy-four younger adults,age from 23 to 50,with femoral neck fractures were treated by open veducition,internal fixa- tion and pedicled bone transplantation from Jan.1995 to Dec.2004.All of the 74 patients were reviewed clin- ically and radiologically after an average of 3.2 years.Results In this group,19/74 cases(25.68%)had avaseular necrosis of the femoral head,which were diagnosised after an average of 28.5 months following inju- ry.Despite these results,these patients assessment with Harris system had been very good or good in 55/74 patients(74.32%).Conclusion It's an effective method to decrease the incidence of necrosis of femoral head after management the femoral neck fracture in younger patients by microsurgical techniques.

Objective To evaluate the clinical effect of restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) for refractory ulcerative colitis (UC).Methods In this study 60 refractory UC patients received IPAA operation during the period of 1990 to 2010.Data were collected regarding early and late postoperation complications,anal continence function,and characteristics of feces.The patients' quality of life was objectively accessed using the Clevend Global Quality of Life (CGQL)index.0ne-way analysis of variance was used.Results Mean follow-up period was 2 years.Early postoperative complication rate developed in 15％ (9/60),including abdominal or pelvic infection,anastomotic leak,pouch bleeding,pouch-vaginal fistula,and intestinal obstruction.Late postoperative complication rate was 12％ (7/60),including pouchitis,intestinal obstruction,and male sexual dysfunction.Stool frequency per 24 hours and that at night was 3.5 ± 1.3 and 1.4 ±0.6.93％ (56/60)patients differentiated gas and feces well and 3％ (2/60) needed daily pads.According to Kirwan Grading Scale,anal function outcomes were Grade Ⅰ:54 (90％),Grade Ⅱ:4 (7％),and Grade Ⅲ:2 (3％).According to Bristol Stool Form Scale,characteristics of feces were Grade Ⅳ:31 (52％),Grade Ⅴ:25(42％),and Grade Ⅵ:4 (6％).Postoperative CGQL result showed a much better quality of life than preoperative CGQL (F =12.368,P ＜ 0.05).Conclusions Refractory UC is surgically indicated and IPAA is the operation of choice with safety,satisfactory long-term outcome and improved quality of life.

OBJECTIVETo observe the effect of Yishen Zhongzi Pill (YZP) on the sperm motility of Shen-essence deficiency syndrome (SEDS) patients.

METHODSTotally 144 patients were randomly assigned to the treatment group (71 cases) and the control group (73 cases). Ten cases dropped out during the therapeutic course. A total of 134 patients completed this clinical study, 67 in each group. Patients in the treatment group took YZP (6 g each time, twice daily), while those in the control group took Wuzi Yanzong Pill (6 g each time, twice daily). The therapeutic course for all was one month. Changes of the sperm motility, clinical efficacy, and Chinese medical symptoms scores were observed between the two groups before and after treatment.

RESULTSCompared with before treatment in the same group, percentages of class a spermatozoa and class a + b spermatozoa were enhanced in the two groups after treatment (P <0. 01). Besides, they were higher in treatment group than in the control group (P <0. 05). The total effective rate of the treatment group was 77. 61 % (52/67 cases), while it was 58. 21 % (39/67 cases) in the control group. It was better in the treatment group than in the control group (Z= -2. 914, P =0. 004). Compared with before treatment in the same group, scores for Chinese medical symptoms decreased in the two groups (P <0. 01). The difference of pre-post integrals was higher in the treatment group than in the control group (P <0. 01).

CONCLUSIONYZP showed favorable effect in treating asthenospermia patients with SEDS and could effectively elevate the motility of sperm.

Asthenozoospermia ; therapy ; Biomedical Research ; Drugs, Chinese Herbal ; Humans ; Male ; Medicine, Chinese Traditional ; Oils, Volatile ; Phytotherapy ; Sperm Motility ; Spermatozoa ; Syndrome

To study the chemical constituents of Dipsacus asper, chromatographic methods such as D101 macroporous resin, silica gel, octadecylsilyl (ODS) column chromatographic techniques and preparative HPLC were used, and five compounds were isolated from 70% (v/v) ethanol extract of the plant. By using spectroscopic techniques including 1H NMR, 13C NMR, 1H-1H COSY, HSQC, HMBC and TOF-MS, the compounds were identified as 3beta-hydroxy-24-nor-urs-4 (23), 12-dien-28-oic acid (1), ursolic acid (2), oleanolic acid (3), 3-O-alpha-L-rhamnosyl(1 --> 3)-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl (1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranosyl ester (4), 3-O-[beta-D-xylopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 4)] [alpha-L-rhamnosyl(1 --> 3)]-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl(1 --> 2)-alpha-L-arabinopyranosyl hederagenin (5), separately. Among them, 1 is a new compound, and 2 is isolated from this plant for the first time.

[ ABSTRACT] AIM: To study the protective effects of cannabinoid CB2 receptor agonist JWH133 on rat acute lung injury induced by paraquat (PQ).METHODS: Male Sprague-Dawley rats (n=72) were randomly divided into 4 groups.PQ group:PQ was administered intraperitoneally at the dose of 20 mg/kg;Low-dose JWH133 pretreatment group ( L-JWH133 group):JWH133 (5 mg/kg, ip) was administered 1 h before PQ exposure;high-dose JWH133 pretreatment group ( H-JWH133 group):JWH133 (20 mg/kg, ip) was administered 1 h before PQ exposure;control group:1 mL sa-line was administered intraperitoneally.Arterial blood, bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected at 8 h, 1 d and 3 d after PQ exposure.PaO2 and the levels of TNF-αand IL-1βin BALF were measured via blood gas analyzer and ELISA, respectively.The pathological changes and lung injury scores were assessed at 3 d after PQ expo-sure.NF-κB and AP-1 protein levels were also determined by Western blotting.RESULTS:The decrease in PaO2 , struc-tural injury of the lung tissues, interstitial pulmonary edema, and the increase in IL-1βand TNF-αin BALF were observed in PQ-treated rats compared with control group.JWH133 pretreatment reduced the degree of lung tissue injury, decreased the levels of IL-1βand TNF-αin BALF and the NF-κB and AP-1 protein expression in the lung tissue compared with PQ group, especially in H-JWH133 group.CONCLUSION:CB2 receptor agonist JWH133 inhibits NF-κB and AP-1 protein expression in the lung tissues, and reduces the secretion of IL-1βand TNF-αin BALF after paraquat exposure, thus atten-uating paraquat-induced acute lung injury.

AIM: To determine the effect of suction duration on thickness and diameter of corneal flap created by microkeratome in porcine eyes in laser in situ kerato-mileusis (LASIK).METHODS: Sixty porcine eyes were randomly assigned to three groups according to different suction durations: group 1 (10 seconds), group 2 (20 seconds), and group 3 (30 seconds). A Moria M2 microkeratome (Moria, France) with a 160μm head was used to create a corneal flap. Corneal flap thickness was measured by automated ultrasonic pachymetry, and the flap diameter was measured by a vernier caliper.RESULTS: The flap thickness of group 1, group 2 and group 3 was 146.05±13.46μm, 157.35±18.95μm and 169.25±21.02μm, respectively. There was a statistically significant difference among three groups (P=0.001). The mean flap diameter in groups 1, 2 and 3 was 8.63±0.19mm, 8.89±0.24mm and 9.06±0.18mm, respectively. A statisti-cally significant difference was found among groups (P<0.01).CONCLUSION: In LASIK in porcine eyes, an increase in suction duration resulted in a thicker and greater flap.

entrate graft can block pathologic process and improve the inclusion of femoral head and increase blood circulation of femoral head, which is beneficial to its recovery.

Objective To investigate the effect of minocycline on isoflurane-induced hippocampal neuroapoptosis and cognitive dysfunction in aged rats. Methods Forty-five male SD rats were randomly assigned into 3 groups ( n=15): blank control group ( group C) , 1.5% isoflurane group ( group I) and 50 mg??kg-1 minocycline+1.5% isoflurane group ( group M+I) . Minocycline was injected intraperitoneally 12 h before the start of anesthesia for group M+I.Group I and group M+I were exposed to 1.5% isoflurane for 4 h, while group C were exposed to 30% O2-70% N2.At the end of anesthesia, five rats in each group were randomized to analyse arterial blood gas. The other rats in each group were sent back to their home cage until they were fully awake.Fourteen days after anesthesia, Morris water maze was used to assess the cognitive function, and then hippocampi of rats were dissected for detection of the expression of cleaved caspase3, Bax and Bcl-2. Results No difference was found in arterial gas analysis among the 3 groups (P>0.05).Compared with group C, the rats in the group I spent more time locating the platform on the third and fourth training days and the time percentage that the rats in group I spent in the target quadrant was much less (P<0.05).However, these changes were reversed in group M+I (P<0.05).The isoflurane-induced increased level of Bax and cleaved caspase3 and decline of anti-apoptotic factor Bcl-2 were restored by minocycline pretreatment ( P<0.05) . Conclusion Minocycline could attenuate cognitive dysfunction induced by isofluranein aged rats.The mechanism is associated with inhibition of hippocampal neuroapoptosis which is increased by isoflurane.

OBJECTIVETo discuss the operative techniques of endoscopic endonasal transsphenoidal surgery and perioperative management for treatment of acute pituitar y apoplexy (PA).METHODS A retrospective analysis was conducted on the clinical data of 51 pathologically diagnosed acute PA after transsphenoidal endoscopic endonasal resection at the TianJin HuanHu hospital between January 2000 and December 2013. There were 28 males and 23 females,with median age of 47 and disease course of 4h-7d.The typical clinical manifestations included headache, visual interference and pituitary dysfunction. CT scan, MRI scan and endocrinological examinations were performed in all cases before operation. Glucocorticoids were used during perioperatve period, The postoperation symptoms and the results follow-up visit after operation were recorded. RESULTS The tumors were totally removed in 42 cases and sub-totally removed in 9 cases without operative death and serious complications. The follow-up period was 1 year to 14 years. Of 47 patients with headaches before the operation, all patients were resolved after the operation; In 38 patients with sight disturbance before the operation,the sight was recovered very well in 34 patients after the operation. The visual field was recovered very well in all patients. Of 28 patients with endocrine disturbance before the operation, 22 were improved in endocrine symptom after the operation. No patient recurred.CONCLUSION The endoscopic endonasal transsphenoidal operation is safe and effective for treatment of acute PA.Appropriate perioperative management is important for the success of operation.