Objective To develop and compare the methods for determining the carrier status in the 18 deleted families of Duchenne and Becker muscular dystrophy. Methods Deletion analysis of the probands was performed by multiplex polymerase chain reaction (PCR) to amplify 9 dystrophin exons described by Chamberlain. Polymorphism linkage analysis was made on DNA with PCR amplification using primers of intragenic short tandem repeat sequences (STR44, STR45, STR49 and STR50), primers of 5′ end (5′DYS II) and primers of 3′end (MZ18, MZ19) in the members of the families. Gene dosage analysis was performed and DQ value was calculated. Results Both of deletions of exons and STR allelic fragments adjacent to the deleted exons were determined in the probands. STR allelic fragments of 6 pairs of heterozygotes, 2 pairs of homozygotes and 11 hemizygotes were detected at those loci in all of the female relatives. 13 female relatives in deleted families were assayed with gene dosage analysis. In 9 /13 female relatives DQ value was in the range of single copy and carrier status was ascertained.Conclusion Repeat sequence polymorphism as well as gene dosage analysis can potentially be used in carrier detection in the deleted families of Duchenne and Becker muscular dystrophy.

100% elevation of PKC activity in the retina, and administration of D ? tocopherol prevented the elevation of PKC activity and diabetes induced decrease of both Na + K + ATPase and Ca 2+ ATPase activities. D ? tocopherol achieved a complete prevention of augmented pericyte and endothelial cell profile areas and basement membrane thickening in the superficial and deep capillaries bed of diabetic retina but had no effect on blood glucose and HbA 1c . Conclusion Diabetes induced histopathological abnormalities are mostly mediated by PKC. D ? tocopherol reduces the ultrastructural lesions in retinal capillary bed induced by hyperglycemia.

OBJECTIVE To analyze the multi-drug resistant genes(genes encoding extended-spectrum ?-lactamases,outer membrane protein gene oprD2,aminoglycoside-modifying enzyme genes,sulfonamides and disinfectant-resistance gene and plasmid-mediated quinolone resistance gene) of Pseudomonas aeruginosa clinical strains.METHODS The drug-resistance to 15 antibiotics was detected by the disc agar diffusion(DAD) and microdilution broth method in 32 P.aeruginosa strains isolated from Jun to Dec 2006.Twenty-eight related drug-resistant genes and outer membrane protein gene oprD2 were examined by PCR method.RESULTS The resistance rate of 32 P.aeruginosa strains to amikacin,ceftazidime,ciprofloxacin,cefepime,cefoperazone/sulbactam,imipenem,meropenem,levofloxacin,piperacillin/tazobactam,aztreonam and piperacillin were 9.4%,25%,59.4%,68.7%,68.8%,78.1%,81.1%,81.3%,84.4% and 94.4%,respectively,and that to others antibiotics were 100%.The detective rate of aminoglycoside-modifying enzyme genes(aac(6′)-Ⅱ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aph(3′)-Ⅵ,ant(3″)-Ⅰ and aac(3)-Ⅰ) were 68.8%,62.5%,21.9%,9.4%,9.4% and 6.3%,and the genes encoding extended-spectrum ?-lactamases(blaFOX,blaIMP,blaVIM and blaDHA) were 37.5% 15.6% 6.3% and 9.3%,respectively.Twenty-two(68.8%) of them were detected in oprD2 and 9(28.1%),but positive in qacE?-sul1 gene.CONCLUSIONS The detective rate of aminoglycoside-modifying enzyme genes,extended-spectrum ?-lactamases genes and the loss rate of outer membrane protein gene oprD2 are high in these 32 multi-drug resistant P.aeruginosa clinical strains.

OBJECTIVE To investigate genes associated with aminoglycosides modification enzymes(AMEs)in Pseudomonas aeruginosa(PAE)isolated from NICU patients.METHODS Drug-resistant genes encoding AMEs such as aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ、aac(3)-Ⅳ,aac(6')-Ⅰ,aac(6')-Ⅱ,aph(3')-Ⅵ,ant(3″)-Ⅰand ant(2″)-Ⅰwere detected bypoly merase chain reaction(PCR)amplification in 36 PAE isolates.The bacteria were identified by ATB.RESULTS The positive rates of aac(6')-Ⅱ,aac(3)-Ⅱ,aac(6')-Ⅰ and aph(3')-Ⅵ genes were 52.8%,47.2%,11.1% and 2.8% of 36 isolates,respectively.Drug-resistant genes encoding AMEs were detected positively in 77.8% of 36 isolates.CONCLUSIONS AMEs genes are present in high percentage of PAE isolated from NICU patients.

Objective To report the technique of functional reconstruction for knee joint following destructive injury and its outcome. Methods In the period from June 1991 to April 2001, microsurgery was done for 4 cases to reconstruct functions of the damaged knees. The operations were combined with transplantation of bilateral latissimus dorsi myocutaneous flaps, emergency transplantation of latissimus dorsi myocutaneous flap, emergency transplantation of useless leg composite tissue and emergency rotation- plasty, respectively. Results All the transplants survived completely. The follow- ups, ranging from 4 to 14 years, revealed normal or nearly normal functions in all the repaired knees. Conclusion Whenever possible, a knee with destructive injury should be repaired and reconstructed, and microsurgery may offer an effective solution.

0.05).There existed a increased trend of the above parameters in group B.(2) Time of weaning from mechanical ventilation was shortened in group A than that in group B(P0.05),but were significant(P0.05).Conclusion Standardized nursing can increase the therapeutic effect on MG by reducing MG crisis and complication incidence,and improving quality of life of MG patients.

Aim: To observe the effects of sarsasapogenin ( SAR) on osteoblasts and osteoclasts cultured in vitro. Methods: Colonal murine calvarial osteoblast-like cell line MC3T3-E1 cells were cultured in vitro. MTT,p-nitropheneye phosphate and tinctorial method of alizarin Bordeaux were used to investigate the effects of SAR on the proliferation, ALP expression, and mineralization tuberculation of MC3T3-E1 cells. Mature osteoclasts were i-solated from the long bone of one-day rat. Meanwhile, marrow cells of mouse bone were cultured with induction of 1,25( OH)_2VitD_3. During the culturing of osteoclasts or marrow cells, SAR of different concentrations was added into the medium. The number of osteoclasts was recognized as tartrate resistant acid phosphatase( TRAP) ( +) multinucleate cells and the resorption lacuna on bone slice were examined with toluidine blue staining. Results: Comparing with the control group, SAR (0.01, 0. 1, 1μg/mL) significanthy increased the proliferation of MC3T3-E1 cells (P <0. 05, P <0. 01). There was no significant difference in the expression of ALP in early pro-liferating MC3T3-E1 cells exposed to SAR of 0.01,0. 1, 1μg/mL, but in the differentiation phase MC3T3-E1 cells, SAR improved ALP activity very significantly if compared with the control group, of which SAR of 1 μg/mL had the most promotion effect(P <0. 01). In addition, compared to the control group, there were, to various ex-tents, increased in the number of mineral nodes in MC3T3-E1 cells after 15day incubation with SAR of different conentrations. Furthermore, no obvious effects of 0.01-1μg/mL SAR on mature osteoclast were observed. But typical osteoclasts were formed when marrow cells were cultured with the induction of 1,25(OH)_2D_3 in medium for 7 days while little or no osteoclasts were induced from marrow cells in the presence of SAR. Conclusion: The results suggest that SAR can effectively promote the proliferation, differentiation and mineralization of osteoblasts cultured in vitro. Besides, SAR can inhibit the generation of osteoclasts from marrow cells.

OBJECTIVE To explore the drug-resistance,the existing forms,genotype,and transfer ways of extended spectrum ?-lactamases(ESBLs) and plasmid AmpC enzyme in Klebsiella pneumoniae.METHODS The drug sensitivity of K.pneumoniae to 17 antibiotics was done by slip-diffusion and microdilution methods.The genotype of two enzymes was assessed by PCR and sequencing.The transfer ways of K.pneumoniae drug-resistant gene were identified by transconjugants-test.RESULTS The ESBLs were mainly produced in 55 cefoxitin resistant K.pneumonia strains.The major genotypes of ESBLs and plasmid AmpC nzyme were CTX-M,MIR and DHA.These genes could be transferred from clinical isolates to recipient bacteria.CONCLUSIONS ESBLs as well as AmpC enzymes are the most important resistance mechanism in K.pneumoniae.The resistance could be transferred through the bacterial conjugation.

OBJECTIVE To study the drug resistance genes in imipenem-resistant Pseudomonas aeruginosa.METHODS Five main drug resistance genes:IMP,VIM,SPM and GIM which are pertinent with metal ?-lactamase and outer membrane protein oprD2 gene were analyzed using PCR method.RESULTS The oprD2 genes in 48 strains of imipenem-resistant P.aeruginosa were negative,and in 14 strains were positive from all 62 strains tested.There were 16 strains with positive IMP type and 5 strains with positive VIM type metal enzyme genes positive,7 metal enzymes producing strains were with negative oprD2 gene.The SPM and GIM metal enzyme genes detected were all negative.CONCLUSIONS The loss of outer membrane protein oprD2 gene is the main mechanism of imipenem resistance in P.aeruginosa in Tai′an area.

Objective To compare the therapeutic effect of conventional treatment and conventional treatment plus external application of Shuangbai Powder for patients with wounded limb injured by venomous snake. Methods One hundred patients bitten by venomous snake were randomized into treatment group and control group, 50 cases in each group. The control group was given conventional treatment including repeatedly washing the wound with hydrogen peroxide, debriding the wound, letting blood and draining toxicity, local blocking with chymotrypsin, wet packing the wounded limb with magnesium sulfate, and injection with anti-venomous serum, tetanus antitoxin, antibiotics, furosemide and energy mixture. The treatment group was given external application of Shuangbai Powder on the basis of the treatment for the control group. Both groups were given the comprehensive nursing of psychological nursing, wound nursing, dietary nursing, defecation nursing and functional nursing. The swelling-subsiding time for the wounded limb and pain scores of visual analogue scale (VAS) in the two groups were compared. Results(1) After treatment, swelling-subsiding time for the wounded limb of the treatment group was shorter than that of the control group, the difference being statistically significant (P < 0.05). (2) After treatment for 4 days and at the end of the treatment, VAS scores of the two groups were decreased compared with those before treatment(P < 0.05), and the decrease of VAS scores in the treatment group after treatment for 4 days was superior to that in the control group (P < 0.05). Conclusion The conventional treatment plus external application of Shuangbai Powder is an effective therapy for patients with wounded limb injured by venomous snake by shortening swelling-subsiding time and relieving pain for the wounded limb .