[Objective]To present the key technique of combining procedures of Chiari's ilium osteotomy,partial synovectomy of the hip joint,femur neck drilling decompression,iliopsoas muscle lengthening and detachment of adductor muscles of the femur for treating Legg-Perthes disease and the long-term results of combination treatment. [Method]A retrospective study of treating children with Legg-Perthes disease was carefully carried out.All patients enrolled were followed up more than 3 years after operation.The results of serial radiographs and physical examination of all patients were reviewed.[Result]The long-term results of 82 children with Legg-Perthes were evaluated according to 5-item and ten score system including clinical presentation,limb shortage,acetabula socket coverage of the femur capital epiphysis,shape of the capital ipiphysis and the intertrochanteric distance.Each item was classified into 3 degrees(0,1,or 2) respectively.Score sum ≥9 was rated as being excellent.The score sum 7 to 8 was evaluated as being good and that reaching 5 to 6 as being fair.The sum

Objective To examine the neuroprotective effect of pulsed magnetic field in a animal focal cere-bral ischemica-reperfusion injury model. Methods Forty-eight Sprague-Dawley (SD) rats were randomized into 3 groups, a sham-operation group, a model group and a pulsed magnetic field group, with rats 16 in each group. Mid-dle cerebral artery occlusion (MCAO) method was employed to establish the focal cerebral ischemia-reperfusion inju-ry model in rats of model group and pulsed magnetic field group. Rats in sham-operation group was subject to the same operation procedure but not underwent ischemia-reperfusion. The infarction volume, histopathological damage and expressions of IGF-1 in ischemic brain tissue were investigated to evaluate the effect of pulsed magnetic field. Results The infarction volume was reduced, histopathological damage alleviated and expressions of IGF-1 in ische-mic brain tissue elevated in the pulsed magnetic field group as compared against the model group (P<0.05). Con-clusions Pulsed magnetic field might provide neuro-protection against cerebral ischemia-reperfusion injury.

Objective To study effect of nasal tolerance with rat-derived 97-116 peptide of AChR α-subunit(Rα97-116(V108A))on the manifestation of muscle weakness and the immunity function of experimental autoimmune myasthenia gravis(EAMG).Methods Twenty-two EAMG model Lewis rats immunized thrice with Rα97-116(V108A)were divided randomly into tolerance group and control group.They were respectively immunized with Rα97-116(V108A)and PBS buffer solution for 10 days via nasal mucous.Then the body weight and Lennon score of two group Lewis rats were measured.Their serum anti-AChR antibodies were tested by ELISA,the expression levels of CD28,CTLA4,B7-1 and B7-2 were determined by flow cytometry.Results Compared with control group at different time points.the body weight of tolerance group rats(tolerance group(228.1±5.8)g,control group(215.0±16.2)g,t=2.395,P＜0.05)increased,the mean clinical score of rats(tolerance group 1.55±0.44.control group 2.10±0.66,t=-2.20,P＜0.05)decreased and the amount of serum anti-AChR antibody(tolerance group 0.97±0.20,control group 1.27±0.26,t=-2.857,P＜0.05)decreased obviously.the amount of CD28,B7-1,B7-2,CTLA4(%)expressed on the surface of peripheral blood cells(tolerance group:27.35±7.05,4.73±0.58,2.71±0.35,1.72±0.44,control group:40.02±8.81,9.52±1.25,5.88±1.09,2.64±0.47)down-regulated markedly(t=3.479,10.861,8.755,4.403,all P＜0.01).Conclusion Nasal mucous tolerance with Rα97-116(V108A)could ameliorate muscular weakness of EAMG rats while activates T cell and inhibits B cellular immunity.

Objective To establish a technical route for the efficient expression and purification of PprI protein from Deinococcus radiodurans R1 by using eukaryotic Pichia pastoris.Methods The encoding sequence of the Deinococcus radiodurans pprI gene was modified according to the preference of Pichia pastoris' codon.Modified pprI gene was fully synthesized with PCR and a 6 × His tag was added at its Nterminal.The PCR products were purified and then cloned into Pichia pastoris expression vector pHBM-905A.After utilizing Cop I and Not I double enzyme digestion and retrievering linear objective fragment,new pprI gene was transformed to the GS115 strain of Pichia pastoris.The obtained Pichia pastoris transformants were induced to express.Culture supernatants were detected by SDS-PAGE,Western blot,and mass spectrometry.A Ni-NTA column was uesd to purify the target protein and the BCA method was used to determine protein concentration.Results The coding sequence of new synthetic Deinococcus radiodurans pprI gene was correct.The purpose protein band of a molecular weight of 43 000 was detected in the culture supernatant of transformed Pichia pastoris strains by SDS-PAGE and Western blot.The mass spectrometry confirmed that it was the Deinococcus radiodurans PprI protein.When the concentration of imidazole was 250 mmol/L,the elution rate of PprI protein was the highest.The purified protein concentration was 0.35 mg/ml measured by BCA method.Conclusions This study has successfully constructed a new pprI gene and the recombinant strain of Pichia pastoris secreting PprI protein,and established a technical route for the efficient expression and purification of PprI protein.

objective To investigate the recurrence of stroke,clinical prognosis and vascular changes in patients with ischemic stroke due to middle cerebral artery stenosis. Methods The ischemic stroke patients with symptomatic middle cerebral artery stenosis were enrolled continuously and followed up prospectively for six months. The recurrence of ipsilateral stroke,clinical prognosis and dynamic changes of vessels were analyzed. Results Eighty patients were included,and 20.0% of the patients(16 cases)presented with recurrence of ipsilateral ischemic stroke and 56 cases (70.0%)with a good outcome(modified Rankin scale[mRS]≤1)during the 6 months follow-up;38.6% patients (27 cases) presented with significant vascular changes with progression in 12 cases (17.1%)and regression in 15 cases (21.4%). Conclusion The patients with simple symptomatic middle cerebral artery stenosis have an high rate recurrence of ipsilateral stroke but have good prognosis;Lesioned artery of the majority of patients in the short period after stroke was stable,but vascular stenosis in some patients could appear progression or remission.

Objective To explore surgical methods of removing polyacrylamide hydrogel (PAHG) and the right time of repairing the deformity of breast after removing PAHG.Methods We operated with endoscopy to remove PAHG through the lower mammary areolar incision.According to injection influence,we made the dicision whether to put the silicon gel prosthesis (hereafter referred to as prosthesis) simultaneously,and to fix the porsthesis,and to rebuild the imframammary fold of breast using biological repair membrane (hereafter referred to as membrane).Results In all 46 patients,39 patients' injection and their envelope were removed entirely.7 patients left part of the envelope because of its thin and wide characters.14 of them accepted prosthesis augmentation mammaplasty simultaneously and 5 of these accepted membrane repair.4 patients received prosthesis augmentation in stage Ⅱ.All patients' incision were primary healing.The incision scars were not obvious.1 patient with breast cancer suffered breast excision; 1 patient who received prosthesis and membrane simultaneously appeared prosthesis displacement after 3 months and fixed again.1 patient who received membrane appeared hydrops in residual cavity,and the membrane was removed finally.Conclusions This method with endoscopy through mammary areolae is necessary for cleaning PAHG entirely.We can use prosthesis to repair the deformity of breast after removing PAHG,and if necessary use membrane to fix the implant and rebuild the inframammary fold of the breast.

ObjectiveTo evaluate the clinical efficacy and safety of astragalus injection on the immune function in patients with senile sepsis.Methods Sixty patients with old age sepsis in Critical Care Medicine Department of Guangdong Provincial Traditional Chinese Medicine Hospital were enrolled and randomly assigned into control and treatment groups according to the table of random numbers, 30 cases in each group. According to 2012 sepsis guidelines for treatment, including antibacterial drug, mechanical ventilation, visceral function support, etc., the therapy was given to the control group; besides the treatment in the control group, intravenous drip of 60 mL astragalus injection(10 mL per ampoule) in 250 mL 0.9% normal saline was additionally given in the treatment group, once a day for 7 days. Before and after treatment, the immunological indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score, sequential organ failure assessment(SOFA) score, duration of mechanical ventilation and time of stay in intensive care unit(ICU), 28-day mortality and adverse drug reactions were compared between the two groups.Results Before treatment, there were no statistically significant differences in CD3+, CD3+CD4+, CD3+CD8+ and T helper cells /T suppressor cells(Th/Ts)levels between the two groups(allP>0.05), while CD3-NK+ of the control group was significantly higher than that in the treatment group〔(10.47±6.22)% vs. (6.26±4.13)%,P<0.05〕. After treatment in treatment group, CD3+, CD3+CD4+ and CD3-NK+ were increased, CD3+CD8+,Th/Ts were decreased compared with those before treatment; in the control group after treatment, CD3+,CD3+CD8+ and CD3-NK+ were decreased and CD3+CD4+ and Th/Ts increased compared with those before treatment. In the comparisons between the treatment group and control group after treatment, the differences in CD3+, CD3+CD4+ and CD3+CD8+ had statistical significance〔CD3+:(30.30±17.17)% vs.(41.91±22.29)%, CD3+CD4+:(31.54±13.24)% vs.(40.08±15.28)%, CD3+CD8+:(14.25±8.10)% vs.(9.52±9.33)%,allP<0.05〕; while the differences in Th/Ts and CD3-NK+ had no statistical significance(bothP>0.05). After treatment in the treatment group, IgG was increased compared with that in the control group〔IgG(g/L): 13.07±5.43 vs. 10.10±3.96,P<
0.05〕. The differences in IgA, IgM, complement(C3,C4) and total serum complement activity(CH50) in the comparisons between the two groups had no statistical significance after treatment(allP>0.05). The differences in APACHEⅡ score(13.83±6.18 vs. 15.90±7.48), SOFA score(7.38±4.66 vs. 6.89±4.19), time of stay in ICU(day: 11.63±5.13 vs. 13.62±8.08), invasive ventilation time(hour: 155.44±119.68 vs. 224.08±174.15) and noninvasive ventilation time(hour: 55.55±42.24 vs. 98.57±43.17) had no statistical significance in comparisons between the treatment group and control group after treatment(allP>0.05). The difference in 28-day mortality had no statistical significance in comparison between the treatment group and control group〔16.7%(5/30) vs. 20.0%(6/30),P>0.05〕. In 60 cases, there were 2 patients with adverse drug reaction, one diarrhea and another little rashes, the rest of the patients did not appear any drug side effect.ConclusionAstragalus injection combined with conventional western medicine therapy possibly has certain effect on adjustment of disturbance of immunologic functions in old patients with sepsis, and its therapeutic safety is well.

0.05). In the group of pirenzepine, the RBF, velocity and volume significantly increased at 0.5 h and 1 h after injection compared with that before injection (P0.05). Conclusion These results suggest that pirenzepine could increase RBF and oxygen in blood with the peak time at 0.5 h and 1 h after intravitreous injection.

Objective To investigate the expression of HSP70 mRNA or MDR1 mRNA by quercetin in leukemia.Methods Methyl thiazoly tetrazolium(MTT) asssy was used to detect the growth suppression of Quercetin on K562 cells.The expression of HSP70 mRNA and MDR1 mRNA were detected in ADM plus Quercetin,Quercetin plus ADM group.The 2 of them were detected in K562/ADM cells in Quercetin plus ADM group.Results 1.Quercetin could suppress the growth of K562 cells.2.The expression of HSP70 mRNA and MDR1 mRNA detected by reverse transcriptase-polymerase chain reaction(RT-PCR) decreased in K562 and K562/ADM cells in Quercetin plus ADM group compared with ADM group and control(P

BACKGROUNDThe role played by the nitric oxide (NO) signaling pathway in the aqueous humor dynamics is still unclear. This study was designed to investigate the expression and distribution of NO synthase (NOS) isoforms and guanylate cyclase (GC) in human ciliary body, trabecular meshwork and the Schlemm's canal.

METHODSTwelve eyes after corneal transplantation were used. Expression of three NOS isoforms (i.e. neuronal NOS (nNOS), inducible NOS (iNOS) and endothelial NOS (eNOS)) and GC were assessed in 10 eyes by immunohistochemical staining using monoclonal or polyclonal antibody of NOS and GC. Ciliary bodies were dissected free and the total proteins were extracted. Western blotting was performed to confirm the protein expression of 3 NOS isoforms and GC.

RESULTSExpression of 3 NOS isoforms and GC were observed in the ciliary epithelium, ciliary muscle, trabecular meshwork and the endothelium of the Schlemm's canal. Immunoreactivity of nNOS was detected mainly along the apical cytoplasmic junction of the non-pigmented epithelium (NPE) and pigmented epithelial (PE) cells. Protein expressions of 3 NOS isoforms and GC were confirmed in isolated human ciliary body by Western blotting.

CONCLUSIONSThe expression of NOS isoforms and GC in human ciliary body suggest the possible involvement of NO and cyclic guanosine monophosphate (cyclic GMP, cGMP) signaling pathway in the ciliary body, and may play a role in both processes of aqueous humor formation and drainage.