Objective To investigate the influence of super fine crushing technique on dissolution rate of Sunshang Capsule (SC). Methods With the dissolution of active components and notoginsengside as the markers, the dissolution rate in Sunshang Capsule and Sunshang powder which were prepared by super fine crushing method and fine crushing method respectively was compared by TLC and HPLC. Results The stain of Sunshang Capsule was bigger and darker than that of Sunshang Powder, time-limited dissolution of SC was 4.66 %higher and the content of notoginsengside R1 was 6 %higher than those of Sunshang Powder. Conclusion Super fine crushing technique can increase the dissolution rate of SC and this will supply evidence for the process of SC preparation.

Objective To establish the preparation method of Xiao' er Jiere hollow type suppository and determination method of the dissolubility.Methods Xiao' er Jiere hollow type suppository and ordinary suppository were prepared.With bacalin and chlorogenic acid as the standards,the in-vitro dissolution test and HPLC method were applied to determine the accumulated dissolution rate of bacalin and chlorogenic acid in two different types of suppository.Results The linearity of bacalin was in the range of 0.040 ? g～ 0.320 ? g(r=0.999 7)and that of chlorogenic acid was in the range of 0.064～ 0.512 ? g(r=0.999 8).The accumulated dissolution rate of the hollow type suppository was over 97 % in 30min.Conclusion Drug release rate of hollow type suppository is higher than the ordinary suppository.The method is stable and accurate.

Objective To investigate the surgical strategy and outcome for occipitocervical extramedullary tumors.Methods 15 patients with occipitocervical extramedullary tumors from January 2007 to July 2010 were reviewed retrospectively.There were 7 males and 8 females with an mean age of 44.6 years (range,21-72 years).All cases were intradural tumors,including 7 cases in dorsolateral,5 cases in lateral side,and 3 cases in ventralis of spinal cord.Functional and neurological statuses were assessed using the Frankel grade and Japanese Orthopaedic Association (JOA) scale.According to Frankel grade system,there were 4 patients with Grade C,8 with Grade D,and 3 with Grade E.The mean preoperative JOA score was 10.5±3.6.Based on the tumor site,there were 5 cases in medulla oblongata-C1 and 10 cases in C1,2.Frankel grade and JOA score were used to evaluate neurological status and general health.The improvement of symptoms and characteristics of surgical strategies were analyzed.Results All cases were followed up for 2-4 years (mean 2.8 years).Complete resection of tumor and good fusion of bone graft were found in radiography postoperatively.The pathological diagnoses included 10 cases of schwannoma,3 cases of meningioma and 2 cases of neurofibroma.There was statistical difference between the preoperative and the final follow-up functional and neurological statuses including Frankel grade and JOA score.At the latest follow-up,2 cases improved from Frankel grade C to D,2from C to E,6 from D to E,but none in 2 cases with grade D.The mean JOA score at final follow-up was 15.6±1.6,and the average improvement rate was 80.7％± 17.4％.The mean preoperative C0-2 angle was 27.0°±4.1 °,and C2-7 angle was 16.8°± 12.7°.C0-2 angle was 27.6°±8.8°,and C2-7 angle was 10.2°±6.8° at the latest follow-up.However,these differences did not reach statistical significance.Conclusion Surgical treatment can effectively maintain or improve neurological function and improve quality of life.Extramedullary tumors can be resected completely by posterior approach,and spinal stabilization can be obtained satisfactorily through selecting appropriate surgical strategies.For medulla oblongata-C1 level,occipitocervical fusion is usually chosen after extirpation of tumor.For C 1,2 level,C1,2 fusion after tumor resection is useful in preventing atlantoaxial instability.

OBJECTIVETo observe the influence of heat shock preconditioning on the expressions of heat shock protein (HSP) 60 and HSP 70 and on the activities of cytochrome oxidase (CCO) and superoxide dismutase (SOD) in mitochondria in gastric mucosa of severely scalded rats, and to investigate its protective mechanism on acute gastric mucosal lesion in rats with severe scald.

METHODSOne hundred and forty-four Wistar rats were randomized into three groups, i. e. scald group ( n = 40, acute gastric mucosal lesion was made after scald, other 8 normal rats without scald were employed as blank control); HS group ( n =40, with heat shock preconditioning 20 h before scald), and other 8 rats preconditioned with heat shock but without scald were employed as experimental control I; actinomycin D group ( n = 40, with intraperitoneal injection of 0. 1 mg/kg actinomycin D 30 min before heat shock preconditioning and other treatment as HS group), and other 8 rats with merely actinomycin D injection were employed as experimental control II. Eight rats in each group were sacrificed and laparotomized at 3, 6, 12, 24 and 48 post-scald hours (PSH) , respectively to determine the index of gastric mucosal lesions (UI ) , the mRNA expressions of HSP70 and protein expression of HSP60 and HSP70, and the changes in the activities of SOD and CCO.

RESULTSUI of the scalded rats increased as the time elapses, reaching the peak (12. 8 +/- 1.9) at 24 PSH. In addition, UI in HS group was significantly lower than that in scald group at each time-point except that at 3 PSH ( P < 0. 05 or 0. 01). The extent of gastric mucosal lesion in rats in actinomycin D group was obviously aggravated compared with that in scald and HS groups ( P <0. 05). The HSP70 mRNA expression in both scald and HS groups was increased at each time-points except for 48PBH, while that in actinomycin D group was increased at 24 PBH and 48PBH. The expressions of HSP70 and HSP60 were greatly increased in HS group compared with those in scald group ( P < 0. 05 or 0. 01) , while those in actinomycin D group were significantly inhibited ( P < 0. 05). The activities of CCO and SOD were gradually decreased in gastric mucosa in scald group, but it was greatly improved by HS preconditioning at 6, 12, 24 PSH ( P < 0. 05 or 0. 01).

CONCLUSIONHeat shock preconditioning is beneficial for the protection of acute gastric mucosal lesion of rats after severe scald, due to increase of HPS60 and HSP70 expression, and increase of CCO and SOD activities in mitochondria.

Animals ; Burns ; metabolism ; pathology ; Chaperonin 60 ; metabolism ; Electron Transport Complex IV ; metabolism ; Female ; Gastric Mucosa ; metabolism ; pathology ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; Male ; Mitochondrial Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

Objective: The detection and analysis of exon mutations of pneumoconiosis and pneumoco-niosis complicated with lung cancer provide reference evidence for screening, clinical diagnosis and treatment and prognosis in pneumoconiosis and pneumoconiosis complicated with lung cancer. Methods: The pathologi-cal tissue samples from 3 pneumoconiosis patients and 3 pneumoconiosis complicated with lung cancer pa-tients were collected. Genomic DNA was extracted and library was prepared. Exomes of the pathological tissue samples in pneumoconiosis patients and pneumoconiosis complicated with lung cancer patients were se-quenced using Ion Torrent PGM platform. Results: Mutation genes FGFR3, PDGFRA, KDR, APC, EGFR, FGFR2, SMO, TP53, RET and CDKN2A were detected in pathological tissue samples of 3 pneumoconiosis patients; Mutation genes FGFR3, PDGFRA, KIT, KDR, APC, EGFR, FGFR2, TP53, RET, CDKN2A, ATM, NPM1, MET and FLT3 were detected in pathological tissue samples of 3 pneumoconiosis complicated with lung cancer patients (P<0.01) . FGFR3, PDGFRA were detected in pathological tissue samples of each pa-tient (mutation frequency>98%) ; Differential genes KIT, ATM, NPM1, MET and FLT3 were only detected in pathological tissue samples of pneumoconiosis complicated with lung cancer patients but not pneumoconio-sis patients. Conclusion A variety of exon mutations detected in pneumoconiosis patients and pneumoconiosis complicated with lung cancer patients using high-throughput sequencing technique have potential value of ap-plications in screening, clinical diagnosis and treatment and prognosis in pneumoconiosis and pneumoconiosis complicated with lung cancer.

Objective To investigate the mechanism and suppression effect of human cytomegalovirus (HCMV) on proliferation of megakaryocyte progenitors(CFU-Mk)in vitro.Methods Colony forming unit-assay was applied to observe the effect of HCMV-AD 169 strain on CFU-Mk of cord blood. The technique of PCR was used to demonstrate the existence of HCMV-AD 169 DNA in the colony cells of cultured CFU-Mk.Results 1.The number of CFU-Mk colonies in HCMV-infected groups decreased significantly compared with that of control group. The CFU-Mk formation was inhibited significantly after HCMV-AD 169 strain infection.The suppression effect showed a dose-dependent fashion: 46.7 % inhibition with 10 -1of HCMV, 29.7 % with 10 -2 and 14.5 % with 10 -3 in the CFU-Mix assay. The peak of CFU-Mk colonies (d16-18) was not significantly different between control group and experimental groups, but the duration of the CFU-Mk colonies in infected groups was significantly shorter than that in control group.2. HCMV-DNA was positively detected in the colony cells of viral infected group by PCR, while negative in control group.Conclusions HCMV-AD 169 strain may inhibit the differentiation and proliferation of CFU-Mk by infecting the hematopoietic progenitors. HCMV may cause the suppression of hematopoiesis by direct infection, which may be the main reason for HCMV infection associated with thrombocytopenia.

OBJECTIVE Bergapten (BG), is a furanocoumarin derived from herbal and citrus extracts can act as antioxidant and selective anticancer agents.The current study aimed to investigate whether bergapten would attenuate immunosenescence and to exploreits immunomodulatory effects on immune responses in D-galactose-induced aging BALB/c mice.METHODS Firstly,mice were given D-galactose(180 mg·kg-1)subcutaneous injections for 30 d.To evaluate the establishment of the aging-related effect in mice, serum samples of BALB/c mice were collected from tail vein. Aging BALB/c mice were freely divided into three groups: negative control group received 1% Tween 80 solution only, named D-gal group. Positive groups were received BG administration at the dose of 20 and 100 mg·kg-1, named D-gal+BG(20)group and D-gal+BG(100)group,respectively.Effects of bergapten on T lympho-cyte proliferation and flow cytometry were assessed by using the splenic cell suspension. Enzyme linked immunospot kits were used to quantitatively determine interferon-γ(IFN-γ)and interleukin-4(IL-4) levels of the isolated serum. Immunophenotype was determined by using mixture of antibodies includ-ing anti-CD3,anti-CD4,and anti-CD8.RESULTS Bergapten(20 mg·kg-1)therapy can modulate immu-nity against viral epidemics and attenuate aging-induced immune deficiency(P<0.01),which was correlat-ed with the decline in the activation of the Th and Tc responses in D-galactose induced aging BALB/c mice.According to the in vivo results,bergapten exposure up-regulated the secretion of IFN-γ and IL-4 in T-helper 1(Th1)and T helper 2(Th2)cells(P<0.05,P<0.01).Additionally,BG(20 mg·kg-1)restored antigen-specific CD4+and CD8+T cells in aging models (P<0.05, P<0.01), which may help to curing chronic infections. CONCLUSION The beneficial effect of bergapten in D-galactose induced aging BALB/c mice may be due to the Th and Tc responses activation.

Objective To investigate the association of angiotensinogen(AGT)gene A-6G、T174M and G-217A polymorphisms with the risk of essential hypertension(EH)in the elderly of Han nationality.Methods Genotypes of AGT gene A-6G,T174M and G-217A polymorphisms in 177 aged EH patients and 86 sex and age-matched controls were analyzed with gene chip technology.Results The A-6G and T174M polymorphisms of AGT gene were significantly associated with EH.The numbers of the three genotypes of A-6G were 113,58 and 6 in the patient group and 70,15 and 1 in the control group(P= 0.014)and those of T174M were 94,77 and 6,60,25 and 1(P=0.031),respectively.G-217A polymorphism was not related to EH.Individuals carrying A-6G AA and T174M CC genotypes showed 57% and 56% lower risk of EH(OR=0.43;95%CI=0.23-0.82 and OR=0.44;95%CI=0.25-0.79, respectively).Conclusions The A-6G AA and the T174M CC genotype may be related with decreased risk of EH and G-217A polymorphism may have little role in the etiology of EH in Han nationality.

OBJECTIVETo investigate the expression of extracellular matrix metalloproteinase induced (EMMPRIN) in the interface tissue, and explore the role of EMMPRIN in the aseptic loosening of prostheses.

METHODSImmunohistochemistry was performed to characterize the EMMPRIN-expressing cells at sites of interface tissue around aseptic loosened hip prostheses in 16 cases. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to study the existence of EMMPRIN mRNA in interface tissue samples. And it was followed up by computer assisted image analysis in order to detect the A values of their expression. Synovium of hip joint of 8 femoral neck fracture were in control group.

RESULTSStrong immunostaining of EMMPRIN was found in the macrophages and fibroblasts of lining-like layers and vascular endothelium of synovial membrane-like interface tissue around loosened prostheses. Expression of EMMPRIN was significantly higher in interface tissue than the control synovium (z=-3.252, P=0.001). RT-PCR of interface tissue samples disclosed the presence of EMMPRIN mRNA of 14 cases. In interface tissue, the A value of EMMPRIN increased significantly compared to control synovium (P<0.01).

CONCLUSIONOver-expression of EMMPRIN up-regulates the production of matrix metalloproteinase (MMPs) in the interface tissue. And it can promote the bone destruction around prostheses. Thereby it may be one of methods to prevent and treat aseptic loosening of prostheses by repression the biology activity of EMMPRIN.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; Basigin ; genetics ; physiology ; Female ; Hip Prosthesis ; Humans ; Immunohistochemistry ; Male ; Matrix Metalloproteinases ; metabolism ; Middle Aged ; Osteolysis ; physiopathology ; Prosthesis Failure ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Synovial Membrane ; metabolism

To optimize the culture environment and protocol of hematopoietic cells' expansion, avoiding the fluctuation caused by medium changing in stirred culture and concentration gradient in static culture, the hematopoietic cells from cord blood (CB) were cultured in a stirred bioreactor connected with a cell retention system, which is a gravity sedimentation settler designed for hematopoietic cell. Total cells expanded 11.5 and 18.6 fold respectively in the twice perfusion stirred cultures, in which CFU-Mix was expanded 23.2 and 20.4 fold, CFU-GM 13.9 fold and 21.5 fold, BFU-E 8.0 fold and 6.9 fold, CD34+ cells 17.1 fold and 15.4 fold. After 12-day culture, it was obtained that 1082 x 10(6) total cells, 6.31 x 10(6) CFU-GM, 6.2 x 10(6) CFU-Mix and 23 x 10(6) CD34+ cells from 267 x 10(6) CB mononuclear cells (MNC) in the first culture, and 1080 x 10(6) total cells, 4.65 x 10(6) CFU-GM, 11.0 x 10(6) CFU-Mix, and 25.0 x 10(6) CD34+ cells from 180 x 10(6) CB MNC. These two cultures met to the clinical scale. Due to the optimized dissolved oxygen (DO) and stable culture environment, the rate of stem/progenitor cells to total cells in the perfusion culture was higher than that in T-flask cell-retention feeding culture. But the cell growth was inhibited in the later phase of perfusion culture, when the cell density is high. The inhibition should be attribute to the high cell density itself. The perfusion culture environment in bioreactor with optimal DO and pH controlling is more favorable for stem/progenitor cells' maintenance and expansion, and the expanded cells' number has reached a clinical scale. But the high cell density in the later phase of perfusion culture caused inhibition to mature hematopoietic cell's growth.
Bioreactors ; Cell Culture Techniques ; methods ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans