BACKGROUND:Bone graft for acetabular reconstruction includes morselized bone graft, structural bone graft and hybrid bone graft, and the morselized bone has been widely used because of the advantages of simple
production and short healing time.
OBJECTIVE:To explore the key technologies and clinical effect of impaction bone grafting with morselized bone in total hip revision for AAOS Ⅲ acetabular deficiency.
METHODS:Sixteen cases of AAOS Ⅲ acetabular deficiency were treated with impaction bone grafting with morselized bone combined with metal devicesor constructive bone grafting. The hip Harris scores and
radiographic data were compared before and after treatment. The effect of impaction bone grafting with morselized bone on acetabular deficiency was assessed.
RESULTS AND CONCLUSION:Al the patients were fol owed-up at 3, 6, 12 months after surgery and every half a year successively. The pain of hip joints after operation was relieved significantly and the walking function was
restored. The hip Harris score was improved from 48.00 points before surgery to 84.94 points after surgery (P<0.01). Five cases were graded as excel ent, eight cases as good, two cases as average, and one case as poor. The excel ent and good rate was 81%, and the satisfying rate of the patients was 94%. The post-operative X-ray films of al the 16 patients showed that the acetabular rotation centers were recovered (near) to normal and the acetabular cups were covered wel by bone. The grafting bone particles got radiological osseointegration and the
acetabular cup prosthesis did not displaced, and no displacement and breakage happened to the metal devices.
Impaction bone grafting with morselized bone in total hip revision for AAOS Ⅲ acetabular deficiency can effectively reconstruct the acetabular bone structure, retain and restore the acetabular bone mass, and it has good technical advantages and good clinical effects.

Corynebacterium glutamicum SA001 is a mutant with lactate dehydrogenase (ldhA) deletion. In order to increase metabolic flux from isocitrate to succinate, and to improve the production of succinate under anaerobic conditions,we transducted the gene aceA coding isocitrate lyase (ICL) from Escherichia coli K12 into Corynebacterium glutamicum SA001 (SA001/pXMJ19-aceA). After 12 h aerobic induction by adding 0.8 mmol/L of IPTG, the recombinant strain was transferred to anaerobic fermentation for 16 h. Succinate reached 14.84 g/L, with a productivity of 0.83 g/(L x h). Compared to C. glutamicum SA001, the activity of ICL of the recombinant strain was increased 5.8-fold, and the succinate productivity was increased 48%. Overexpression of isocitrate lyase will increase the metabolic flux of glyoxylate bypass flowing to succinate.
Corynebacterium glutamicum ; genetics ; metabolism ; Escherichia coli ; enzymology ; genetics ; Gene Deletion ; Industrial Microbiology ; Isocitrate Lyase ; biosynthesis ; genetics ; L-Lactate Dehydrogenase ; genetics ; Succinic Acid ; metabolism ; Transduction, Genetic

Objective To develop an efficient recombinant expression system of alanine aminotransferase ( ALT ) in order to build the foundation for the preparation of feedstock related ALT reference materials.Methods A new human ALT gene was synthesized by optimizing the codons of the nucleic acid sequence encoding human ALT using bioinformatic tools, and then it was cloned into pRSF-Duet expression vector.The recombinant plasmid pRSF-Duet-ALT was transformed into E.coli BL21 and the target protein expression was induced by 2 mmol/L isopropyl β-D-1-thiogalactopyranoside ( IPTG ) .The expression condition for soluble protein was optimized by changing the inducer concentration, shaking speed and induction temperature. The soluble protein was purified by nickel ion affinity chromatography and dextran molecular sieve chromatography, and identified by sodium dodecyl sulfate-polyacrylamide gelelectrophoresis( SDS-PAGE) and Western blot analysis.The activity and stability of recombinant proteins in serum matrix under different storage conditions were detected.Results The usage frequency in E.coli of ALT codons was more than 10%after codon optimization.The expressions of soluble proteins were increased by optimizing the induced expression conditions, including a final concentration of 2 mmol/L of IPTG, and continued incubation with shaking at 150 rpm for 8 h at 25℃.The purified protein was identified as ALT by SDS-PAGE and Western blot with ALT activities of up to 80 000 U/L.Recombinant ALT could be stable for 2-8 d at 2-8 ℃or 25 ℃with a relative standard deviation of less than 5%.Conclusions An efficient recombinant expression system of ALT was developed successfully by codons optimization.The obtained recombinant protein could achieve the requirements of reference material feedstock.

Objective To establish a method for determining the dissolution oftorasemide sustained-release tablet in vitro and study the methodology of the determination.The consistency of the in vitro release behavior between self-prepared torasemide sustained-release tablet and original preparation were evaluated by constructed method.Methods HPLC method was applied to detect the cumulative release percentage of self-prepared torasemide sustained-release tablet and original preparation in five kinds of release media (water,0.1 mol/L hydrochloric acid solution,pH 4.5 acetate buffer,pH 6.8 phosphate buffer,and 0.1 mol/L hydrochloric acid solution turn to pH 6.8 phosphate buffer).Similarity factor (f2) was used to evaluate the similarity of release curves.Results There was a good linear relationship between the quality concentration of torasemide and peak area in the range of 1.0-12.0 μg/mL (r =0.9995).Results of precision and stability tests were good,and the RSDs for probational liquid were all lower than 2.0％.The average recovery of accuracy test was 100.04％,and RSD was 0.54％ (n =12).The homogeneity of within group of self-prepared preparation met the technical requirement,RSDs of each sampling points in six Dissolution Vessels were lower than 10.0％.The f2 factors of self-prepared torasemide sustained-release tablet and original preparation were 72,60,77,66,and 60 in five kinds of release media.Conclusion The method in the paper is suitable for the release test of torasemide,meanwhile,the self-prepared tablet shows consistent in vitro release behavior with that of the original preparation.

Objective Surgical treatment of lung cancer needs to follow strict quality control standard for the aims of accuracy of pathological staging and potentially improved prognosis.However,there are short of studies related to surgical quality analysis.Methods One hundred and twelve patients were enlisted with the diagnosis of lung cancer and received surgical intervention in 2007.Surgical quality of these cases were retrospectively analyzed in compliance with three international standards,National Comprehensive Cancer Network (NCCN),American College of Surgeons’ Oncology Group (ACOSOG),and International Association for the Study of Lung Cancer (IASLC).Results No surgical death was reported in this group.According to NCCN,ACOSOG and IASLC standards,qualified operations were 82 cases (80.4％),73 cases (71.6％),45 cases (44.1％) in 102 cases with R0 resection,respectively.The median total lymph nodes,median mediastinal nodes,and N1 nodes were 27 (range 0-63),16.5 (range 0-43),and 10.5 (range 0-26),respectively.The median mediastinal node stations resected were 4 (range 0-6).In the mediastinum,stations of 4R,5,6 and 7 presented the highest frequency of receiving lymph node dissection.For N1 stations,10 and 12 were among the top list.For the whole group,1-year survival,3-year survival and 4-year survival were 92％ (95％ CI,89-95),67％ (95％ CI,62-72),57％ (95％ CI,52-62),respectively.Conclusion Quality control is the essential part of surgical treatment of lung cancer,which will facilitate the baseline standardization of clinical research.Since IASLC provides the strictest standard for lung cancer surgery,we suggest that all thoracic surgeons need to follow this standard to secure the accuracy of pathological diagnosis and for a potential better prognosis.

Objective To investigate the species composition and quantitative dynamic state of rodent and parasitic flea in Huairou District of Beijing City,and to carry out serologic monitoring of animal plague.Methods Rodent density was investigated at different habitats in eight villages and towns between 2009 and 2011.Serum samples from captured rodents were detected by indirect hemagglutination test,and genotypes of collected parasitic fleas were identified.Results A total of 709 rodents which belong to eight different kinds were captured with a rodent density of 2.52％ (83/3 300),and Rattus niviventer and Apodemus peninsulae were dominant species,acounted for 46.4％ (329/709) and 18.1％(128/709),respctively.Five hundred and sixty-three serum samples of rodents were tested by plague indirect hemagglutination,and all of them were plague antibody negative.Two hundred fleas were checked with a flea infected rate of 29.63％ (48/162),and a flea index of 1.23; Paraceras crispus and Leptopsylla pavlovskii were dominant flea species,a total of 30 and 27,respectively.Conclusion Some environment conditions are suitable for rodents survival,even though there is no sign of animal plague epidemic.

OBJECTIVBE To investigate the intervention of compound Astragalus and Salvia milt-iorrhiza extract (CASE) consisted of astragalosides, astragalus polysaccharides and salvianolic acids on the interaction of microRNA-145/microRNA-21 (miR-145/miR-21) and Smad3C/3L phosphorylation (pSmad3C/pSmad3L) down-stream of transforming growth factor-β (TGF-β)/mitogen activated protein kinase (MAPK) signaling in hepatocellular carcinoma (HCC) progression by in vitro and in vivo experi-ments. METHODS In HepG2 cells and xenografts of nude mice, antagomir/agomir and plasmids of Smad3C/3L phosphorylation site mutation (Smad3 3S-A/Smad3 EPSM) were used to intervene miR-145/miR-21 and pSmad3C/pSmad3L expression respectively,then incorporative CASE treatment. Cell proliferation, migration, apoptosis, tumor growth and histopathologic characteristics of xenografts, relevant proteins of TGF-β/Smad pathway and miR-145/miR-21 were evaluated.RESULTS CASE up-regulated miR-145 while down-regulated miR-21, inhibited cell proliferation,migration and tumor growth, accelerated cell apoptosis in HepG2 cells respectively transfected with Smad3 WT, Smad3 EPSM,Smad3 3S-A plasmids in cultured dishes and xenografts of nude mice,the above effects were more evident in HepG2 cells with increased pSmad3C.In TGF-β1-stimulated HepG2 cells and xenografts of nude mice, CASE antagonized the facilitating effects of miR-145 antagomir/miR-21 agomir on cell migration,proliferation,tumor growth and inhibiting effects of miR-145 antagomir/miR-21 agomir on cell apoptosis; CASE increased miR-145 down-regulated by miR-145 antagomir and decreased miR-21 up-regulated by miR-21 agomir,reduced protein level of pSmad3L and their proteins including TβRⅡ, pERK1/2, pJNK1/2 and pp38 while elevated pSmad3C expression. CONCLUSION These results suggest that pSmad3C/pSmad3L maybe interact with miR-145/miR-21 in HCC progression,which may be one of important molecular mechanisms of CASE's anti-HCC effects.

OBJECTIVETo observe the clinical efficacy difference in treatment of functional dyspepsia (FD) between syndrome differentiation based acupuncture and ordinary acupuncture.

METHODSSeventy FD patients were assigned to a syndrome differentiation based acupuncture group (Group A) and an ordinary acupuncture group (Group B) by Excel Software randomization. Zhongwan (RN12 ), Tianshu (ST25), and Zusanli (ST36) were needled as main points for patients in Group A. Meanwhile, different combined acupoints were needled according to syndrome differentiation. Only the same main points were needled for patients in Group B. All patients were needled once per day, 30 min each time, 6 days as one treatment cycle, 2 treatment cycles in total. Fasting serum levels of gastrin (GAS) and motilin (MTL) were determined before treatment and after 2 treatment cycles. 36-item Short-form Heath Survey (SF-36) and Nepean Dyspepsia Index [NDI, including Nepean Dyspepsia Symptom Index (NDSI) and Nepean Dyspepsia Life Quality Index (NDLQI)] were assessed before treatment, after 2 treatment cycles, and one month after treatment.

RESULTSCompared with before treatment in the same group, serum levels of GAS and MLT increased in the two groups after 2 treatment cycles (P <0. 05), but changes were more obvious in Group A (P <0. 05). Compared with before treatment in the same group, SF-36 and NDLQI score increased, and NDSI score decreased in the two groups after 2 treatment cycles and 1 month after treatment (all P <0. 05). Compared with Group B, SF-36 and NDLQI score increased in Group A after 2 treatment cycles and 1 month after treatment (P <0. 05, P <0. 01). But NDSI score at 1 month after treatment was lower in Group A than in Group B (P <0.01).

CONCLUSIONSyndrome differentiation based acupuncture could evidently improve dyspeptic symptoms of FD patients, and significantly improve their quality of life with remarkable curative effect.

Acupuncture Points ; Acupuncture Therapy ; Dyspepsia ; therapy ; Humans ; Motilin ; Needles ; Quality of Life ; Surveys and Questionnaires ; Syndrome

To explore the molecular mechanism of human papillomavirus subtype 16(HPV-16)E7 oncogene-induced DNA re-replication in response to DNA damage. Flow cytometry was performed to examine the cell cycle changes in RPE1 E7 cells stably expressing HPV-16 E7 and its control cell RPE1 Vector after DNA damage.Immunoblotting assay was used to evaluate the early mitotic inhibitor 1(Emi1)expression in RPE1 E7 and RPE1 Vector cells with or without DNA damage.The changes of the proportion of polyploidy was detected by flow cytometry in DNA-damaged RPE1 E7 cells interfered by Emi1 small interfering RNA. Compared with the control cells,the proportion of polyploids in RPE1 E7 cells was significantly increased in response to DNA damage(=6.397,=0.0031).Emi1 protein expression was significantly increased in DNA damaged RPE1 E7 cells(=8.241,=0.0012).The polyploid ratio of RPE1 E7 cells was significantly reduced after Emi1 was interfered by two independent small interfering RNAs(=2.916,=0.0434;=3.452,=0.0260). In response to DNA damage,Emi1 promoted DNA re-replication caused by HPV-16 E7.
DNA Damage ; DNA Replication ; Human papillomavirus 16 ; Mitosis ; Oncogene Proteins, Viral

OBJECTIVEThis study aims to detect microRNA-17(mir-17) expression on the osteogenic differentiation of advanced glycation end products (AGEs)-stimulated hunman periodontal ligament stem cells (HPDLSCs) and to analyze the influence of these cells on this process.

METHODSHPDLSCs were isolated using limited dilution technique. After osteogenic differentiation occurred, different time points of mir-17 expression in the experimental groups were detected by real time polymerase chain reaction (PCR). The mir-17 overexpression and inhibition were evaluated using cell transfection technique. Differences in gene expressions were detected by real time PCR; differences in protein expressions were analyzed by Western blot.

RESULTSThe mir-17 expression was reduced after osteogenic differentiation occurred at 3, 7, and 14 d compared with that in the control group (P < 0.05). The expression levels of bone sialoprotein (BSP), Runt-related transcription factor-2 (Runx-2)and alkaline phosphatase (ALP) in the experimental groups were lower than those in the mimic control group when mir-17 expression increased. In addition, the protein expression levels of Runx-2 in the experimental groups were lower than those in the control group. The expression levels of BSP, Runx-2 and ALP in the experimental groups were higher than those in the inhibitor control group when mir-17 expression decreased. Likewise, the protein expression levels of Runx-2 in the experimental groups were higher than those in the control group.

CONCLUSIONAGEs inhibit the osteogenic differentiation of HPDLSCs by affecting mir-17 expression.

Alkaline Phosphatase ; Cell Differentiation ; Glycation End Products, Advanced ; Humans ; MicroRNAs ; Osteogenesis ; Periodontal Ligament ; Stem Cells