OBJECTIVETo explore the effect of silencing protein kinase D (PKD)-2 on Tca8113 cell proliferation, programmed cell death, and chemosensitivity.

METHODSThe stable cell lines of pkd-2 gene silencing and empty vector plasmid group were established. The proliferation and 50% inhibitory concentration (IC50) of shRNA-mediated Tca8113 chemotherapy drugs were detected through methyl thiazolyl tetrazolium assay (MTT). The programmed cell death rate and sensitivity to Tca8113 chemotherapy drugs before or after pkd-2 gene silencing were measured through flow cytometry. P-glycoprotein (P-gp) expression of pkd-2 silencing cells was identified by immunohistochemical methods.

RESULTSStable cell lines of pkd-2 gene silencing were established. Compared with parental cells, the proliferation of shRNA-mediated Tca8113 was not significantly different, but its IC50 was lower. Meanwhile, cell programmed death rate and sensitivity to chemotherapeutic drugs of shRNA-mediated Tca8113 significantly increased. Compared with wild group Tca8113, a significant decrease in P-gp expression was induced by chemotherapy drugs with shRNA-pkd-2 gene silencing.

CONCLUSIONThe pkd-2 gene of shRNA interference silencing Tca8113 promotes programmed cell death of Tca8113, reduces the IC50 of the chemotherapy drugs, and significantly improves the sensitivity of Tca8113 to chemotherapeutic drugs while reducing the expression of P-gp.

Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Gene Silencing ; Genetic Vectors ; Humans ; Protein Kinase C ; RNA, Small Interfering ; Transfection

Objective:To study the influence of harvest time on the characters of Chaenomeles Spciosa ( Sweet) Nakai to determine the optimal harvest time with traditional characters of the Chinese medicine. Methods: In order to explore the optimal harvest time of Chaenomeles Spciosa ( Sweet) Nakai, the content of alcohol-soluble extract, weight and acidity of Chaenomeles Spciosa ( Sweet) Nakai with different harvest time were determined, the weather conditions in recent 3 years was summarized, and the drying process was also studied. Results:The average weight of Chaenomeles Spciosa(Sweet)Nakai was the lowest in June and highest in August, and the den-sity reached maximum in mid-July. During the whole harvest time, the content of alcohol-soluble extract was stable. The weather con-ditions from mid-July to late-July were with high temperature, dry and little rain, which was suitable for drying of Chaenomeles Spciosa ( Sweet) Nakai. Conclusion:The optimal harvest time of Chaenomeles Spciosa( Sweet) Nakai is confirmed in mid-July according to the traditional customs, drying conditions and characters of the Chinese medicine.

OBJECTIVE To analyze the contamination situation in the oxygen aspiration facility and the measure for infection control.METHODS The first half of the year 2006 was determined as a traditional disinfection supervision group,the second half of the year 2006 was an improvement disinfection group,65 and 62 unused damping bottles in traditional disinfection and improvement disinfection groups were detected.Sixty three and 62 samples of continuously used damping liquid in the two disinfection groups were detected daily.The damping liquid detection of d1 and d6 in above two groups were compared each with other.The detection result was statistically analyzed.RESULTS The statistical analysis of the unused damping bottle qualification ratio between the two groups was with significant differences,P

Objective IFN-? is a pleiotropic cytokine with potent immunomodulatory effects and antiviral activity. To study the mechanism of IFN-? clearing duck hepatitis B virus (DHBV) in ducks, it is essential to establish a method to quantify expression of DuIFN-? in immune response. In the present study,a semi-quantitative competitive RT-PCR was developed to quantify expression of duck IFN-?(DuIFN-?) mRNA by PBMCs. Methods Based on ?-actin consensus sequence, fishing the ?-actin gene as house keeping gene from duck PBMC by RT-PCR. A competitive internal control was constructed and the competitive RT-PCR system could be used to quantify the transcription of DuIFN-? mRNA. Results After duck PBMCs were stimulated in vitro with PHA, the peak of DuIFN-? expression was at 24-36h. Then RT-PCR method was applied to detect DuIFN-? mRNA transcription by PBMCs from DHBV infected ducks immunized with DuIFN-? plasmid plus DNA vaccine or DNA vaccine alone. Results showed that expression of DuIFN-? in ducks co-immunized with DuIFN-? plasmid were higher than other groups immunized without DuIFN-? plasmid as adjuvant. Conclusions The results indicated that DuIFN-? gene could be a useful adjuvant to develop vaccines. The semi-quantitation of DuIFN-? mRNA by competitive RT-PCR provides the basis for future study of the mechanism of IFN-? in duck hepatitis B virus persistent infection.

Xuedang(Ardsia punctata Lindl)is a traditional medicine of Yao nationality commonly used in Guangxi.Plant resources,indication,macroscopic appearance,microscopic char8cteristics,and TLC identification of thecrude drug were studied in comparison witb the easily confusable A. orrota to provide referencial informa-tions for clinics,quality control,development and identification of the Yao medicine.

Objective　To analyze the CT features and diagnostic valuation of endometriosis. Methods 　The CT features in 26 cases with endometriosis confirmed by operation and pathology were retrospectively studied. Results　The endometriosis deposited in the overly （n=16）,wall of bladder （n=2）,uterus （n=5）,and Vagina （n=3）.On CT,the lesion could appear as a round or oval or irregular,Cystic or solid or mixed mass,with unclear margin,and certain adhesion to surrounding sssstructures.There were some small cysts within the mass.On enhanced CT,the cystic margin and solid part of the mass could be enhanced. Conclusion　CT features with clinical information,it is possible to diagnosis endometriosis.

BACKGROUND: Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Researches suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.OBJECTIVE: To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 (IL-1),IL-6 and tumor necrosis factor alpha (TNF-α) in aqueous humor of rabbits after extracapsular cataract extraction.DESIGN: Randomized controlled animal study.SETTING: Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.MATERIALS: The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes,general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification: SYXK (yue) 2005-006]. Eyes of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawei Biotechnology Company Limited.METHODS: Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mi/kg urethan, and then, 5# needle was punctured from corneal limbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque: horizontally intercepting bladder or waterly separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1st, 3rd, 7th and 14th days after operation, inflammatory reaction of anterior chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meanwhile, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.MAIN OUTCOME MEASURES: Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rabbits in three groups after operation.RESULTS: ① On the 1st, 3rd, 7th and 14th days after operation, numbers of leucocytes were (2.4±0.7)×106/L, (2,2±0.5)×106/L, (2.8±0.8)×106/L and (2.0±0.5)×106/L in aqueous humor; (19.7±7.3)×106/L, (28.1±9.6)×106/L, (14.2±5.6)×106/L and(8.4±3.8)×106/L in traumatic control group; (65.3±14.5)×106/L, (79.8±12.7)×106/L, (21.7±8.2)×106/L and (12.4±4.1)×106/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group (F =22.5, 27.9, 11.6, 8.4;P＜0.05). ② Within 1-14 days after operation, contents of IL-1, IL-6and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group (P＜0.05), and there was a significant difference between traumatic control group and operative group (P＜0.05);however, there was no significant difference among three groups on the operative day (P＞0.05). ③ Contents of cytokines reached peak on the 7th day after operation, decreased gradually, and reached the lowest value on the 14th day.Contents in traumatic control group and operative group were higher than those in normal control group (P＜0.05).CONCLUSION: The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.

Objective To study the effects and mechanisms of resveratrol (Resv) on the expression of adiponectin receptors ( AdipoR1 and AdipoR2 ) in rat mesangial cells (MCs) cultured in intermittent high glucose medium.Methods The MCs cultured in normal glucose ( 5.6 mmol/L) medium were transfected by the plasmid vector of the FoxO1 short hairpin RNAs ( FoxO1 shRNA ),and then cultured in intermittent high glucose medium (5.6mmol/L or 30 mmol/L,alternately every 8 hours ) and resveratrol ( Resv 20 μmol/L).The MCs cultured in normal glucose (5.6 mmol/L) medium served as normal glucose control group (NG),and the MCs exposed to iutermittent high glucose medium were divided into five groups:intermittent high glucose group (IHG),IHG+Resv group,IHG+FoxO1 shRNA group,IHG+ Resv + FoxO1 shRNA group,IHG + Negative control group ( shNC ),each group was cultured for 72 h.The level of reactive oxygen species (ROS) was assessed by Fluorescence microplate reader.The mRNA levels of Sirt1,Foxo1,AdipoR1,and AdipoR2 were assessed by RT-RCR.The protein levels of FoxO1,phosphorylation FoxO1 ( p-FoxO1 ),AdipoR1,and AdipoR2 were assessed by Western blotting.Results ( 1 )Compared with NG,intermittent high glucose significantly decreased the mRNA expression of Sirt1,but markedly increased the level of p-FoxO1.Furthermore,the mRNA and protein expression levels of AdipoR1 were obviously decreased while the level of ROS was enhanced in IHG ( all P＜0.05 ).(2) Compared with IHG,the mRNA and protein expression levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was enhanced in group IHG+FoxO1 shRNA ( all P＜0.05 ).( 3 ) Compared with IHG,after treating with Resv,the mRNA expression level of Sirtl was increased,whereas the level of p-FoxO1 was decreased.Moreover,the mRNA and protein levels of AdipoR1 was increased while the level of ROS was lowered in group IHG+Resv (all P＜0.05 ).(4) Compared with group IHG+Resv,the mRNA and protein levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was increased in group IHG+Resv+FoxO1 shRNA (all P＜O.05).(5) In addition,the mRNA and protein expression level of AdipoR2 showed no significant difference among these groups ( P＞0.05 ).Conclusion ( 1 ) Resveratrol significantly increases the expression of AdipoR1 in MCs induced by intermittent high glucose.( 2 ) FoxO1 plays an important role in regulating the expression of AdipoR1 by resveratrol.

Streptozotocin-induced diabetic rats were treated with resveratrol for 12 weeks.Compared with group of diabetic rats without treatment,the levels of urine albumin,serum creatinine,and blood urea nitrogen were significantly decreased and pathological changes in kidney were improved in treatment group ( P＜0.05 ).The mRNA expressions of FoxO1 and catalase were higher and FoxO1 phosphorylation level in diabetic rats treated with resveratrol was lower than that in diabetic rats without treatment (P＜0.05),suggesting that resveratrol may protect the kidneys of diabetic rats via regulating expression of FoxO1.

Objective To investigate the relations between intrauterine asphyxia and peroxidation and newborn hypoxic-ischemic encephalopathy(HIE). Methods The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in cord blood of 60 newborns with intrauterine asphyxia during labor(which was divided into two groups,39 cases with asphyxia in groupⅠ, and 21 cases with asphyxia in groupⅡ),and in 30 newborns without intrauterine asphyxia(control group) were determined. The levels of SOD and MDA in cord blood of newborns with HIE were compared with those in newborns without HIE. The incidence of HIE was estimated simultaneously. Results (1) The levels of SOD were (12 896?247) U/g Hb in groupⅠ, (9846?268) U/g Hb in groupⅡ, (17 282?134) U/g Hb in control group, significantly lower in the former two groups compared with control group, while the level of SOD in group Ⅰ was higher than that in group Ⅱ(P121 min group, and the levels of SOD was (9786?249) U/g Hb.(2)The levels of MDA were (6.3?0.4) ?mol/L in group Ⅰ, (8.6?1.5) ?mol/L in group Ⅱ, and (4.1?0.5) ?mol/L in control group, significantly higher in the former two groups compared with control group (P