Objective To evaluate the therapeutic efficacy of intraoperative mitomycin C(MMC) and TobraDex ophthalmic ointment(TDO) in transcanalicular Nd:YAG laser assisted dacryocystoplasty(LDCL). Methods A total of 204 patients (246 eyes) with lacrimal duct obstruction were divided into four groups: group treated with MMC, group treated with TDO, group treated with MCC plus TDO and control. Prospective control study was conducted. Results The healing rates of three drug groups were 88.7%, 88.6%, 89.5%, respectively, significantly higher than that of the control group (73.1) ( P

Objective To evaluate the therapeutic efficacy of KTP Nd: YAG laser assisted dacryocystoplasty for lacrimal duct obstruction. Methods Specially made hollow lacrimal probe was used to examine the lacrimal ducts to the obstructed parts in a total of 827 eyes. Then KTP laser fibers were inserted into the lacrimal ducts to dredge the obstructed parts by laser. Instillation of different drugs or short term placement of lacrimal supporters was conducted after operation according to the symptoms such as degree of obstruction and with discharges or not. Results The cure rate, improvement rate, and ineffectiveness rate were 93.5%, 5.5%, and 1%, respectively. One therapy was conducted in 562 eyes, twice in 191 eyes. Conclusion KTP Nd: YAG laser assisted dacryocystoplasty is a safe and effective way for the treatment of lacrimal obstruction. Postoperative drug instillation or short term placement of lacrimal supporter can contribute to the clinical outcomes.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

This paper overviewed typical government compensation sources and practices for public hospitals in the world.Government compensation should be made based on regional health planning,while the central government shoulders greater compensation responsibility.The fee-for-case mix is found to be the best incentive The government adjusts its funding baselines for different hospitals to adjust the compensation.In view of the compensation for public hospitals in China,the paper analyzed the enlightenments and lessons from international experiences The authors recommend an evolution from the pattern of compensation per person/per bed,to payment by service unit or volume(for example,per outpatient or emergency visit and days of stay),and in the end to that of payment per disease.

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

ObjectiveTo discuss the correlation of the surgical risk score with the change of T cell subsets and the occurrence of postoperative complications. MethodsA total of 260 patients with hip fractures treated in our department were enrolled in this study and divided into high-risk group ( Group A) and low-risk group (Group B) based on the surgical risk score. The fasting peripheral venous blood was taken in the morning at one day before surgery and at days 1,3, 5, 7 and 14 after surgery for measuring CD3, CD4 and CD8 levels respectively in two groups.In the meantime, the correlation of level changes with risk score and postoperative complications was observed in two groups. ResultsThere were two patients with lung infection in the Group B, with no death. There were two patients with pulmonary infection, one with wound infection and two with deep vein thrombosis, with one death. The postoperative levels of CD3 and CD4 in the Group A and Group B were significantly lower than those in the control group (P ＜ 0.01 ), which reached the lowest level at day 1 after operation and recovered to normal at day 5 after operation. The postoperative levels of CD3 and CD4 in the Group A recovered near to normal at day 7 and to normal at day 14. While the postoperative levels of CD3 and CD4 in the Group B remained low level even at day 14. The level of CD8 decreased at days 1 and 5 in the Group A, then increased and remained relatively stable, while the level of CD8 increased in the Group B. The T cell subsets in both groups recovered from low to high trend at days 1-7 after surgery. The higher preoperative score had more obvious decrease and slower recovery of the T cell subsets. ConclusionsSurgical risk score has positive correlation with the change of T cell subsets and postoperative complications, which can more accurately predict the postoperative outcome of the old patients.

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To investigate the role of Ephrin A2 in the regeneration and reinnervation of hair cells in the chick cochlea following kanamycin ototoxicity.Methods 66 newly hatched Roman chickens (3 days old) were randomly divided into experimental group and control groups. Experimental chickens (n=48) received intramuscular kanamycin (200 mg/kg:Sigma,St Louis,MO) for 10 consecutive days and were subsequently sacrificed 2 days before the last injection,and 1,3,7,10,15,21,30,and 60 days after the last injection (n=6 per subgroup). Control chickens (n=18) were untreated and sacrificed 3,13 and 43 days after hatching (n=6 per subgroup). Ephrin A2 protein expression in acoustic ganglia was determined by western blot analysis in all chickens after sacrifice. Results Ephrin A2 protein expression was found and the protein level was almost same in acoustical ganglia of all normal chickens. After kanamycin exposure,the Ephrin A2 protein expression level in the cochlea of the experiment chickens from 2 to 7 days after last kanamycin injection was lower than that in control chickens,respectively. Ephrin A2 expression increased obviously at 15 days after kanamycin last injection. By 30 days after the cessation of kanamycin treatment,the level of Ephrin A2 protein approximated to that in normal control group.Conclusion The expression of Ephrin A2 protein in the acoustical ganglia basically synchronizes with the regeneration and the reinnervation of the hair cells in the chicken cochlea following kanamycin damage,indicating that Ephrin A2 may play an important role in this process.

Objective To investigate the relationship between bed rest time and development of prethrombotic state in the elderly patients with hip fracture.Methods One hundred and sixty-six patients who stayed in bed after hip fracture,aged ≥65 yr,were divided into 5 groups according to the bed rest time on admission to hospital:bed rest time ＜ 24 h (Ⅰ group,n =61),bed rest time 24-48 h (Ⅱ group,n =29),bed rest time 3-6days (Ⅲ group,n =29),bed rest time 7-14 days (Ⅳ group,n =34),and bed rest time ＞ 14 days (Ⅴ group,n =13).Venous blood samples were collected to measure the platelet count,coagulation function,and concentrations of plasma D-Dimer and serum α-granule membrane protein 140 (GMP-140).The development of abnormality in each index was recorded.The development of deep vein thrombosis in both lower extremities was diagnosed using color Doppler ultrasound in D-Dimer-positive patients.Results Compared with group Ⅰ,the abnormal rate of fibrinogen (Fib) and D-Dimer and serum GMP-140 concentrations were significantly increased in Ⅱ and Ⅲ groups,the abnormal rate of platelet count,Fib and D-Dimer and serum GMP-140 concentrations were increased in lⅣ group,and the abnormal rate of platelet count was increased,and no significant change was found in the serum GMP-140 concentrations and abnormal rate of Fib and D-Dimer in Ⅴ group.Compared with group Ⅱ,the serum GMP-140 concentrations were significantly increased in Ⅲ and Ⅳ groups,the abnormal rate of Fib and D-Dimer was increased in Ⅳ group,and no significant change was found in the abnormal rate of Fib and DDimer in Ⅲ group.The abnormal rate of platelet count was significantly lower in Ⅳ group than in Ⅲ group.Color Doppler Ultrasonography showed no sign of deep vein thrombosis.Conclusion For the elderly patients with hip fracture,the possibility of pre-thrombotic state developed is increased when the bed rest time is more than 24 h,and the patients were classified as high-risk patients when the bed rest time is more than 3 days.

In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.
Aristolochia ; classification ; genetics ; Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Quality Control