Objective To construct the recombinant co-expression vector carrying antisense RNA to dual-target BAG-1 and Bcl-2 genes and recombinant single expression vector carrying antisense RNA to target BAG-1 and Bcl-2 gene respectively,then to pre-liminarily investigate their effect on the proliferation of gastric cancer cell SGC-7901 in order to lay the foundation for further study the effect of this recombinant vector on the tumor cells.Methods RT-PCR was used to amplify the full length of BAG-1 and Bcl-2 cDNA from total RNA of gastric cancer cell line SGC-7901.The BAG-1 cDNA fragment and the Bcl-2 cDNA fragment were insert-ed into pMD18-T simple vector respectively.The pMD18-T-BAG-1 was digested with BamH Ⅰ and Cla Ⅰ and the pMD18-T-Bcl-2 was digested with EcoR Ⅰ and Nhe Ⅰ.Then the BAG-1 cDNA fragment and the Bcl-2 cDNA fragment were inserted into the mcs1 and mcs2 of the eukaryotic co-expression vector pVITRO2 in the antisense orientation respectively.The construction of the single expression vector pVITRO2-AsBAG-1 and pVITRO2-AsBcl-2 was confirmed by restriction endonuclease treatment and se-quence identification.Then the Bcl-2 cDNA fragment was inserted into the mcs2 of the recombinant vector pVITRO2-AsBAG-1 in the antisense orientation to construct the co-expression vector pVITRO2-AsBAG-1-Bcl-2,and the recombinant vector was also iden-tified by restriction endonucleases digestion and sequence identification.Then the recombinant vector was transfected into SGC-7901 cell respectively.The proliferation of the cell was determined by the MTT assay.The level change of BAG-1 and Bcl-2 mRNA in SGC-7901 cell was detected by the semi quantitative RT-PCR and the change situation of cell cycle was detected by the flow cytom-etry(FCM).Results The restriction endonucleases digestion and sequencing identification indicated that the eukaryotic co-expres-sion vector pVITRO2-AsBAG-1-Bcl-2 and the single gene expression vector pVITRO2-AsBAG-1 and pVITRO2-AsBcl-2 were con-structed successfully.The MTT assay method demonstrated that compared with the control group,the recombinant vector could in-hibit the proliferation of the cells in time dependent manner,and the inhibiting effect was most notably in the 72 h transfection group(P <0.01);inhibition ratio of recombinant vector groups was significantly higher than that of control group and pVITRO2 group(P <0.01).BAG-1 and Bcl-2 mRNA expression in recombinant vector groups were significantly decreased compared with that in the control group and pVITRO2 group(P <0.01),The RT-PCR results showed that the expression level of BAG-1 mRNA and Bcl-2 mRNA was significantly decreased(P <0.01),moreover the co-expression vector group was more notably than the single expression vector groups(P <0.05),but the pVITRO2-AsBcl-2 had no obvious effect on the expression of BAG-1 mRNA(P >0. 05);the FCM detection results showed that the apoptosis rate of the recombinant vector groups was significantly higher than that of the control group and the pVITRO2 group with statistical difference(P <0.01),and the co-expression vector group was more nota-bly than single expression vector groups(P <0.01).In addition,the effect of the co-expression recombinant vector group was more significant than that of the single expression co-expression vector(P <0.01 ),the apoptosis rate was increased from 0.57% to 15.75%.Conclusion The co-expression recombinant vector pVITRO2-AsBAG-1-Bcl-2 and single expression vector pVITRO2-As-BAG-1 and pVITRO2-AsBcl-2 are successfully constructed and they can inhibit the proliferation of SGC-7901 cell and induce cell apoptosis,moreover the pVITRO2-AsBAG-1-Bcl-2 vector is most notably.

Objective: To observe the clinical effect of tuina for cervical intervertebral disc herniation. Methods: Eighty seven cases were randomized into a treatment and a control group. 39 cases in the treatment group were treated with tuina manipulation, while 48 cases in the control group were treated with combined cervical traction and medium-frequency impulse. Cases in both groups were treated 3 times a week, and 10 times constitute a course of treatment. Results: Of 39 cases in the treatment group, 22 obtained recovery, 13 obtained marked effects and 4 obtained improvement, manifesting a marked effect rate of 89.7% and an effective rate of 100.0%. Of 48 cases in the control group, 23 obtained recoveries, 17obtained marked effects, 7 obtained improvements and 1 saw no improvement, manifesting a marked effect rate of 83.3% and an effective rate of 97.9%. There were no statistical differences in the marked effect rate and total effective rate. Cases in the treatment group were treated 17.8 times on average, while cases in the control group were treated 26.6 times on average. Conclusion: Despite the equivalent effects in the treatment of cervical intervertebral disc herniation, tuina is superior to combined traction and medium-frequency impulse in terms of a faster effect and shorter course of treatment.

AIM:To study the sedation, diuresis and anticoagulation effects of Ji-Ming-San (JMS) in animal models for exploring the circadian variations of the dosing action and verifying the rationality of administering JMS on special time. METHODS: The sedation effect and circadian difference were determined by observing the time session of ambulation, raising double-forefoot test and autonomic activity of model mice. The diuretic effect and the circadian variation in rats were examined by metabolic cage test while the anticoagulation and the circadian change were observed by glass test. RESULTS: JMS produced significant sedation effect by reducing time session of ambulation, raising upper limbs frequencies/time and autonomic activity and the built-in rhythm with autonomic activity disappeared after JMS administration. JMS had satisfactory diuretic effect and total urinary output in water-loaded rats was increased after dosing. The diuretic effect of dosing showed a circadian rhythm with more significant output during the night than that during the daytime. Also, JMS prolonged the clotting time significantly and the action exhibited circadian difference. As compared with administration at the night, the clotting time was more prolonged at the daytime. CONCLUSION: JMS can produce obvious sedation, diuresis and anticoagulation effects with varied circadian rhythm. The findings suggest that the effect of administering JMS is better at the end of rest phase than other time session.

AIM:To observe effects of Jiming Powder(Semen arecae,Pericarpium citri reticulatae,Fructus Chaenomelis,Fructus Evodiae,Folium Perillae,Radix Platycodonis,and Rhizoma Zingiberis recens) on anticoagulation and hemorheology and on the circadian variations in order to verify the rational practice of timing administering the drug by our ancestors. METHODS:The effect of Jiming Powder on clotting time(CT) was observed by using glass-tube method in mice. Quick test was used to determine the prothrombin time(PT) and circadian variations in rats. The experimental animal models of blood stasis syndrome were prepared by subcutaneous injection of adrenaline in combination with ice water bath for observing the effect of Jiming Powder on hemorheology in such types of animals,and the maximum rate of platelet aggregation was determined by method of Born turbidimetry. RESULTS:Jiming Powder could significantly delay CT and PT characteristic of circadian rhythm. As compared with the administration at daytime,CT and PT were greatly prolonged during the night. Besides,the hemorheological parameters were improved markedly in blood,plasma and fibrinogen viscosity,which suggested that the drug administration was more effective during the night than daytime. Also,the findings showed that Jiming Powder could prevent platelet aggregation induced by adenosine diphosphate. CONCLUSION:Jiming Powder plays a role in anticoagulation and improves the indication of hemorhelogy in the blood-stasis rats model,showing dosing time-dependent.

OBJECTIVE:To determine the titer of main component in kitasamycin tablets and acetylkitasamycin dry suspension.METHODS:Staphylococcus aureus was used as test organism and which were cultured at (37?0.5) ℃ for about 4～5 hours by adding 1.0%～1.5% kitasamycin and 2.0%～2.5% acetylkitasamycin,then the absorbance was determined at a wavelength of 530 nm.RESULTS:The linear range of kitasamycin was 0.5～4.0 IU?mL-1 and that of acetylkitasamycin was 0.45～6.3 IU?mL-1,and their recovery rates were 101.7%(RSD=1.9%,n=9) and 101.1%(RSD=2.1%,n=9),respectively.CONCLUSION:The established turbidimetry is sensitive,rapid yet with few influencing factors,thus it is applicable for the titer determination of main component in kitasamycin tablets and acetylkitasamycin dry suspension.

Objective To evaluate the efficacy of nasendoscopic resection of the inner wall of the maxillary sinus for the treatment of inverted papilloma of the nasal cavity and paranasal sinuses.Methods From January 2002 to December 2006,10 patients with nasal inverted papilloma received nasendoscopic resection of the inner wall of the maxillary sinus in our hospital.The clinical data of the patients were anlayzed retrospectively.Results The operation was completed in all the 10 cases.They were follwed up for 12 to 72 months with a mean of 30 months.Only one of the patients had recurrent papilloma in 1 year after the treatment.None of the patients devleoped epiphora.Conclusions Nasendoscopic resection of the inner wall of the maxillary sinus is feasible for patients with nasal inverted papilloma.By using the procedrue,facial scars can be avoided;moreover,owing to a clear surgical field,the surgeons can completely resect the tumor,and the patients have less trauma and low recurrence rate after the treatment.

Stomatology is an applied science,and experimental teaching plays an important role in training renovation talents.A complete construction of the laboratory system is a complex systematic project,which requires the right teaching conception,advanced experimental instruments,high-quality teachers of laboratory,the optimization of the teaching contents,as well as highly efficient operating mechanism and management approach.It is also a process of exploration and innovation.

Objective To observe teletherapy and local hyperthermia effects on VX2 carcinoma rabbits'cytokines and curative effect. Methods 60 male white health rabbits were selected, and divided them into three group. The group of teletherapy were 20. The group of local hyperthermia were 20. So were the group of comparison. Their tumors' changes, rates of death and changes of IL-2, IL-6, IL-8, TNF, IGF, CA-50 before and after VX2 carcinoma replanted and those treated after one, three, four weeks were observed. Results The diameters of tumors and rates of death from large to small in turn was comparison's group, hyperthermia's group and teletherapy's group. After VX2 carcinoma replanted IL-2, IL-8 were low but IL-6, TNF, IGF, CA-50 were high. After being treated one, three or four weeks, IL-2, IL-8 were getting lower and lower but there was significant difference between teletherapy's group with other group in IL-2 and IL-8(P

[Objective]To explore the effect of combined therapy of Chinese and western medicine on Bell paralysis.[Methods] Randomly divide 62 cases of Bell paralysis into 2 groups who were treated respectively with combined therapy of Chinese and western medicine and western medicine.[Results] For effective rate,the treatment group(91.1%) was markedly higher than control group(67.8%),there was obvious difference between the 2 groups.[Conclusions] Combined therapy of Chinese and western medicine has marked therapeutic effect on Bell paralysis.

OBJECTIVE:To establish a method for simultaneous residual determination of dichloromethane and ethyl acetate in bisacodyl raw material. METHODS:Head-space GC was performed on the capillary column of 6% cyanopropyl phenyl-94% di-methyl polysiloxane(DB-624)by temperature programming,the temperature of injector was 220 ℃,detector was flame ionization detector with temperature of 250 ℃,carrier gas was high purity nitrogen with the flow rate of 3.0 ml/min,split ration was 1∶10, headspace heating temperature was 70 ℃,equilibration time was 30 min,volume of headspace vial was 5 ml,and the injection volume was 1 ml. RESULTS:The linear range was 6-120μg/ml for dichloromethane(r=0.999 9)and 50-1 000μg/ml for ethyl ac-etate(r=0.999 9);the limit of quantitation was 0.2,1.7 μg,limit of detection was 0.06,0.5 μg;RSDs of precision,stability and reproducibility tests were no higher than 3%;recoveries were 100.30%-102.00%(RSD=0.63%,n=9) and 100.10 %-101.30%(RSD=0.44%,n=9). CONCLUSIONS:The method is simple and accurate,and can be used for the simultaneous residual deter-mination of dichloromethane and ethyl acetate in bisacodyl raw material.