To investigate the effect of proteasome inhibitor MG132 on the apoptosis of bovine lens epithelial cells (BLECs), the cells were treated with MG132 at different concentrations for12, 24 and 36 h. The cell viability was analyzed by MTT assay and the effect of MG132 on the apoptosis of BLECs was assessed by flow cytometry (FCM). The results showed that after treatment for the same period, the inhibitory effect of MG132 on BLECs proliferation was enhanced with the increment of the concentration of MG132 (0, 2, 5, 10, mumol/L) (P<0.05). The 50% inhibiting concentration (IC(50)) was 2.03 mumol/L when the BLECs were treated with MG132 for 36 h. MG132 also induced the apoptosis of BLECs obviously. FCM showed that the apoptosis index of the cells treated by MG132 at 2 mumol/L for 12 h was (20.24+/-1.51)%, and that of the control was (0.98+/-0.20)% respectively (P<0.01, n=3). It was concluded that MG132 could lead to apoptosis of BLECs. The decrease of proteasome activity may play an important role in the formation and development of cataract.

Objective To study the effect of various methods determining lung volume and fraction dose on the lung dose-volume parameters for lung cancer patients. Methods Twenty patients with lung cancer were rantdomly enrolled into our study and the plan of three dimensional eonformal radiation therapy(3DCRT)was designed by Varian Eclipse TPS.The lung volumes and the dose-volume parameters were measured under CT value of-300- -980, -400- -980 and -500- -980.Under CT value of -400- -980,total lung volumes were confirmed.The dose-volume parameters of V30,V20,V10 and mean lung dose(MLD)were reevaluated after GTV,CTV and PTV were subtracted from the total lung volumes and when the fraction dose was elevated from 2.0 Gy to 10.0 Gy. Results When the CT value ranged from-300--980 to-500--980,the median reduction of the total lung volumes(-9.10%)was significantly higher than that of V30,V20,V10 and MLD(-3.18%,-1.13%,0.82%and-0.79%,respectively).When the total lung volume was fixed at CT value of-400--980,the alterations of V30,V20,V10 and MLD became more apparent as the increase of the subtracted lung volume,among which the alteration of V30 was most significant while V10 the least.Among five cases with a fixed total dose of 60 Gy and PTV less than 140 cm3,the V30,V20,V10 and MLD were increased to a similar extent(about 40%)when the fraction dose was increased from 2.0 Gy to 10.0 Gy.MLD was increased(36%)when the fraction dose was above 6.0 Gy. Conclusions When CT value ranges from-300- -980 to-500- -980,the total lung volume is influenced most.The alteration of V30,being statistically significant,might have some significance but is not enough to determine the plan of radiotherapy clinically.The alteration of V20、V10 and MLD is not statistically significant.When the overlapped target volume is subtracted from the total lung volumes,the alteration of V30 is the most sign:tifhcant while V10 the least.The fraction dose,being the most consuming factor(＞10%)when comparing with the CT valHe and targeted volume,can significantly influence the dose-volume parameter.

Animals ; Apoptosis ; radiation effects ; Electromagnetic Fields ; adverse effects ; Hippocampus ; pathology ; radiation effects ; Male ; Maze Learning ; radiation effects ; Rats ; Rats, Wistar

OBJECTIVETo investigate the mechanism of retinoic acid (RA) signal in dental evolution, RA is used to explore the influence of the mechanism on neural crest's migration during the early stage of zebra fish embryos.

METHODSWe divided embryos of wild type and transgenic line zebra fish into three groups. 1 x 10(-7) to 6 x 10(-7) mol x L(-1) RA and 1 x 10(-7) mo x L(-1) 4-diethylaminobenzaldehyde (DEAB) were added into egg water at 24 hpf for 9 h. Dimethyl sulfoxid (DMSO) with the concentration was used as control group. Then, antisense probes of dlx2a, dlx2b, and barxl were formulated to perform whole-mount in situ hybridization to check the expressions of the genes in 48 hpf to 72 hpf embryos. We observed fluorescence of transgenic line in 4 dpf embryos.

RESULTSWe obtained three mRNA probes successfully. Compared with DMSO control group, a low concentration (1 x 10(-7) mol x L(-1)) of RA could up-regulate the expression of mRNA (barx1, dlx2a) in neural crest. Obvious migration trend was observed toward the pharyngeal arch in which teeth adhered. Transgenic fish had spreading fluorescence tendency in pharyngeal arch. However, a high concentration (4 x 10(-7) mol x L(-1)) of RA malformed the embryos and killed them after treatment. One third of the embryos of middle concentration (3 x 10(-7) mo x L(-1)) exhibited delayed development. DEAB resulted in neural crest dysplasia. The expression of barxl and dlx2a were suppressed, and the appearance of dlx2b in tooth was delayed.

CONCLUSIONRA signal pathway can regulate the progenitors of tooth by controlling the growth of the neural crest and manipulating tooth development

Animals ; Branchial Region ; Cell Differentiation ; drug effects ; Embryo, Nonmammalian ; drug effects ; embryology ; metabolism ; In Situ Hybridization ; Neural Crest ; drug effects ; Odontogenesis ; Signal Transduction ; Tooth ; drug effects ; embryology ; metabolism ; Tretinoin ; pharmacology ; Zebrafish ; embryology ; genetics ; metabolism

OBJECTIVEThis study aims to investigate the expression of connexin 43 (cx43) gene during early development in zebra fish and provide a foundation for further research of cx43 gene in tooth development.

METHODSTotal RNA was extracted within 72 h after fertilization of zebra fish embryos and then reversed transcribed to generate the cDNA library. The specific fragments of the cx43 gene were then cloned and connected to the PGEMT vector. After confirming the constructed plasmid, the corresponding RNA polymerase was chosen, and the digoxin-labeled anti-sense mRNA probe of cx43 was synthesized in vitro. The cx43 gene expression of zebra fish indifferent stages was carried out by in situ hybridization. The relationship of the cx43 gene expression and anatomy of the pharyngeal teeth were compared by alizarin red staining.

RESULTSThe mRNA antisense probe of cx43 was acquired. The positive signal of sepia was observed in the different stages of zebra fish pharyngeal teeth after fertilization. After fertilization for 9 days, the expression site of cx43 in situ hybridization was overlapped in accordance with the anatomical site of the pharyngeal teeth.

CONCLUSIONcx43 gene participates in tooth development and mineralization process and plays a crucial role in later mineralization.

Animals ; Connexin 43 ; Gene Expression ; Genetic Vectors ; In Situ Hybridization ; Odontogenesis ; Plasmids ; RNA, Messenger ; Tooth ; Zebrafish

ObjectiveTo analyze and summarize the feasibility and characteristics of the posterior spinal canal reduction and fixation in treating mid-upper thoracic spine facture.MethodsA retrospective study was made on 17 patients with mid-upper thoracic spine facture to record the complication, compare the functions of the patients with complete and incomplete spinal cord injuries before and after surgery and examine the iatrogenic injury in patients without spinal cord injuries.ResultsNo complication happened after surgery.Incomplete injury was found in six patients, whose ASIA scales were found to be increased for 1-3 levels during the follow-up.While the complete injury was found in eight patients,whose ASIA scale remained unchanged during the follow-up.The sensory scores of both the incomplete injury group and complete injury group were processed with variance analysis and the results showed a significant difference between pre-operation and post-operation (F = 476.47, P = 0.000).The mean value between complete injury group and incomplete group was with high statistical difference (F = 31.46, P =0.000).The variance analysis of the motor scores showed a significant difference between before and after operation (F=46.75, P =0.000) and the mean value between complete and incomplete injury groups was with statistical difference (F = 158.59, P = 0.000).There were three patients with normal spinal cord function, with no decrease of ASIA scale or no change of the sensory and motor scores.ConclusionsFor patients with mid-upper thoracic spine fracture, posterior spinal surgery is conducive to the recovery of spinal function, for it can safely and effectively avoid worsening the thoracic and other combined injuries and release spinal pressure including the pressure in front part of the spinal cannal.

Objective To observe the bacteriostasis of herba houttuyniae liquid in bladder irrigation for treatment of rabbit bladder infection model, and to offer a new way for prevention and treatment of urinary tract infection.Methods Twenty male New Zealand long-eared white rabbits were divided into gentamicin control group and three different concentrations herba houttuyniae groups according to random number table method, each group 5 rabbits. The rabbit bladder infection model was replicated by injecting coli liquid 1 mL (105 cfu/mL) into the urinary bladder through a urethral catheter, and clipping the catheter for 1 hour. After model establishment for 3 days, in the three herba houttuyniae groups, 0.25, 0.50, 1.00 g/mL herba houttuyniae liquid 50 mL were injected into the bladder respectively, and in the gentamicin control group, 160 kU gentamicin was injected into the bladder. Before molding and after molding for 3, 5, 7, 9 days, ear blood in rabbit was taken, and white blood cell (WBC) count and neutrophil percentage (N%) were tested; at the same time, the middle segment morning urine was taken to detect bacteriuria and urine WBC positive rates; after the experiment, the animals were sacrificed, the urinary bladder tissues were taken, and the pathological changes in urinary bladder tissue were observed by a light microscope.Results Before molding, the bacteriuria in animals of each group was negative and the results of urine WBC, blood WBC and N% were within normal range. After molding for 3 days, bacteriuria and urine WBC positive rates reached 100%; with the prolongation of treatment after molding, the above two rates in each group were gradually decreased, and herba houttuyniae 0.50 g/mL and 1.00 g/mL groups appeared statistical significance on the 5th day (bacteriuria positive rate was 60% and 40%, urine WBC positive rate was 60% and 40% respectively, while the gentamicin control group and herba houttuyniae 0.25 g/mL group was still 100% positive, allP < 0.01); on the 9th day, the bacteriuria positive rate in both higher dosage herba houttuyniae groups was 0, in 0.25 g/mL group, it still had the positive rate 20%, showing herba houttuyniae 0.50 g/mL and 1.00 g/mL had stronger inhibitory effect than herba houttuyniae 0.25 g/mL. WBC and N% were increased in each group after molding, and on the 3rd day, WBC and N were significantly higher in each group than those before molding [WBC (×109/L): gentamicin control group 13.00±5.23 vs. 8.95±2.69, herba houttuyniae 0.25 g/mL group 14.52±10.28 vs. 8.73±3.16, herba houttuyniae 0.50 g/mL group 12.97±11.33 vs. 7.83±4.51, herba houttuyniae 1.00 g/mL group 13.03±9.17 vs. 6.45±3.85. N%: gentamicin control group 80.99±14.93 vs. 63.41±7.96, herba houttuyniae 0.25 g/mL group 89.62±9.41 vs. 70.01±9.13, herba houttuyniae 0.50 g/mL group 87.69±11.34 vs. 67.75±8.91, herba houttuyniae 1.00 g/mL group 85.67±12.81 vs. 65.12±4.63]. On the 7th day, WBC and N% fell, and on the 9th day, the WBC and N% were restored to normal range in herba houttuyniae 0.50 g/mL and 1.00 g/mL groups [WBC (×109/L): herba houttuyniae 0.50 g/mL group 8.56±5.38, herba houttuyniae 1.00 g/mL group 7.93±4.82. N%: herba houttuyniae 0.50 g/mL group 65.97±8.35, herba houttuyniae 1.00 g/mL group 65.57±9.12]. Under light microscopy, it was shown that the morphological appearances of tissue cells in various layers of bladder mucosa were normal in herba houttuyniae group, demonstrating that herba houttuyniae fluid in bladder did not cause allergic inflammatory reaction, etc.Conclusion The bacteriostatic action of herba houttuyniae liquid for bladder irrigation in treatment of rabbits with urinary tract infection is better than that of gentamicin.

Objective To investigate the value of the expressions of estrogen receptor(ER)and progesterone receptor(PR)in the cast-off cells from the fluid of latex ductal lavage in the diagnosis and treatment of breast diseases.Methods A total of 58 female out-patients were examined by fiberoptic ductoscopy(FDS).All patients were divided into 6 groups by the complaints,physical examination,ultrasound and FDS:normal group(10 persons without symptoms and abnormalities in any examination),galactostasis group(n=19),latex fill up group(n=11),intraductal papilloma group(n=2),mammary ductal ectasia group(n=7)and contralateral examination after mastectomy for breast cancer group(n=9).A total of 168 ducts were successfully examined by FDS,and the expressions of ER and PR in the cast-off cells were detected among all of the samples.Results The positive rates of ER and PR expression in the cast-off cells from the fluid of latex ductal lavage in benign breast disease groups were much higher than those in normal group(P

AIM:To study the effect of transient receptor potential channel 1 ( TRPC1) on the survival of hip-pocampal neurons in mice.METHODS:TRPC1 knockout mice and the control mice (6 months old) were used in this study.Immunofluorescence staining of neuron-specific marker NeuN, Nissl staining and TUNEL staining were performed to measure the changes of the neurons in hippocampal CA1, CA3 and dentate gyrus (DG).Western blot analysis was used to detect the levels of pro-apoptotic protein C/EBP homologous protein ( CHOP) and cleaved caspase-3.RESULTS:Immuno-fluorescence staining and Nissl staining showed that the number of neuronal cells was significantly decreased in hippocampal CA1, CA3 and DG of TRPC1 knockout mice compared with the control mice.TUNEL staining showed that the apoptosis neuronal cell number of the above areas in TRPC1 knockout mice was significantly increased compared with the control mice.The results of Western blot revealed that the levels of CHOP and cleaved caspase-3 were significantly increased in the hippocampus of TRPC1 knockout mice relative to the control mice.CONCLUSION:The depletion of TRPC1 induces neu-ronal loss through a mechanism of TRPC1-mediated apoptosis.

ObjectiveTo collect and analyze the literatures on resistance exercise in rheumatoid arthritis (RA),and to evaluate its effectiveness on functional recovery and disease activity.MethodsThe literatures,published on resistance exercise in RA patients indexed by Medline,Embase,OVID and other databases through January 2012,including unpublished literatures,were collected.All literature were enrolled based on the criterion of inclusion and exclusion.Randomized controlled trials (RCTs) comparing resistance exercise with interventions without resistance training for RA patients were included. Outcome values withdrawn from literatures were pooled by the STATA 12.0 software.The heterogeneityand data quality were checked and categorized forMeta-analysis.The mean difference(MD) and 95％ confidence interval (CI)were calculated.ResultsA total of 10 related papers including 525 patients were included in this analysis.Resistance exercise improved isometric strength,isokinetic strength and grip strength.Moreover,disease activity score(DAS) [WMD=-0.585,95％CI (-0.878,-0.291)],erythrocyte sedimentation rate (ESR) [WMD=-3.418,95％CI(-5.289,-1.546)],morning stiffness [WMD=-24.675,95％CI(-29.829,-19.520)],Ritchie joint index[WMD=-2.793,95％CI(-4.682,-0.905)],visual analog scale(VAS) [WMD=-1.412,95％CI(-2.646,-0.177)],health assessment questionnaire (HAQ) [WMD=-0.177,95％CI(-0.345,-0.009)],50-foot walk test [WMD=-2.539,95％CI (-4.260,-0.817)] were significantly different comparing with the control group.With respect to safety,no exercise related adverse events were found in any of the included studies.ConclusionBased on the evidence,as a safe rehabilitation therapy,resistance exercise can not only improve muscle strengh and physical ability,but can also decrease the disease activity levels of RA.