AIM: To observe the effect of combined goniosynechialysis in treating absolute glaucoma after trabeculectomy.
METHODS:Phacoemulsification combined goniosynechialysis was performed on 16 patients ( 16 eyes ) with absolute glaucoma after trabeculectomy, and they were followed up for 6 ~12mo, The postoperative intraocular pressure ( IOP ) and anterior chamber depth, preoperative and postoperative medication types (quantity), preoperative and postoperative 1 month's status of anxiety and depression, symptoms of ocular surface were observed. RESULTS: The IOP decreased significantly after phacoemulsification combined goniosynechialysis. The mean IOP was 35. 00±15. 43mmHg preoperatively, and it was 12. 00±6. 69mmHg, 15. 00±4. 26mmHg and 15. 3±5.2mmHg on 1d, 6 and 12mo after the surgery. The statistic difference was found between preoperative and postoperative (t=6. 22, P<0. 05). The anterior chamber depth was 1. 45 ± 0. 19mm before the surgery, and increased to 3. 37±0. 13mm after the surgery (t=6. 65, P<0. 05). After the surgery, 2 patients needed two kinds of drugs, 2 patients needed one kind of drug. After 12mo of follow-up, anxiety and depression status were improved in all 16 patients. Subjective discomfort symptoms of 16 patients such as eye bilges, eye pain were relieved. All of the patients' eyeballs were preserved, and no serious complications.
CONCLUSION: Phacoemulsification combined goniosynechialysis in treating absolute glaucoma after trabeculectomy is a safe and effective surgical option.

OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

Objective To compare the infectivity between laboratory adapted human inununodefi- ciency virus(HIV-1) and primary HIV-1 isolates for different mucosal epithelial cell lines. Methods Mu-cosal epithelial cells Caco-2, T-84, HeLa and lymphocyte MT-4 were infected with laboratory adapted HIV-1 SF33 and 2 primary HIV-1 isolates (02010561, 02010141). Culture supernatant and cells were collected respectively on 3-4 days interval after virus inoculation. The former was tested for HIV-1 antigen P24 level and viral load, and the latter was tested for total viral DNA and integrated viral DNA. Results All 3 virus strains could infect MT-4 cells and integrate into their genome. Only HIV-1 SF33 could infect Caco-2 cells but could not integrate into their genomic DNA. Both HIV-1 SF33 and 02010561 infected HeLa cells but only integration of HIV-1 SF33 was detected. All the 3 HIV-1 strains infected T-84 cells but only the integra-tion of HIV-1 SF33 and 02010141 was observed. Conclusion Although laboratory adapted and primary HIV-1 strains are able to infect human mucosal epithelial cell lines, transient or productive infection estab-lished in different mucosal epithelial cells is dependent on the character of cells and virus strains.

Objective To investigate the function of myeloid dendritic cells (mDCs) from severe aplastic anemia ( SAA ) patients in stimulating allogeneic T lymphocytes proliferation in vitro and then explore the immunopathogenesis of SAA. Methods Twenty-five SAA patients ( 15 untreated and 10 recovered after immunosuppressive therapy) and 12 normal controls were enrolled in this study. Their mature mDCs were induced from their bone marrow monocytes with recombined human interleukin-4 ( rhIL-4) , recombined human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and recombined human tumor necrosis factor (rhTNF) in vitro. Then mDCs were co-cultured with allogeneic lymphocytes (mixture lymphocyte reaction, MLR) at a ratio of 1: 100 or 1: 50. The growth rate of lymphocyte was measured with methyl thiazolyl tetrazolium ( MTT) colorimetry.The concentrations of interleukin( IL) -12 and inlerferon -y (IFNγ) in MLR supernatant were measured with EL1SA. The correlation between the growth rate and the concentration of IL-12 or IFNγ was analyzed. Results When mDCs and lymphocytes were co-cultured at the ratio of 1: 100, the growth rates of lymphocytes stimulated with mDCs from untreated, recovered SAA patients and controls were (219. 8 ±94. 0)% , (159. 1 ±66. 0)% and (160. 1 ±91. 9)% respectively. The concentrations of IL-12 in MLR supernatant were (8. 2 ± 3. 6) ng/L, (6. 5 ± 2. 8) ng/L and (6. 1 ± 2. 6) ng/L and the concentrations of IFNγ were (21. 8 ± 8. 7) ng/L, (25. 5 ± 9. 1) ng/L and (22. 6 ± 7. 8) ng/L respectively. All of them had no statistical differences among the three groups ( P > 0. 05 ). When mDCs and lymphocytes were co-cultured at the ratio of 1: 50, the growth rate of lymphocytes stimulated with mDCs from untreated patients was (322. 1 ± 171. 1)% , which was higher than that of recovered patients [ (180. 9 ±79. 1)% and controls (192. 3 ±91. 9)% ]. The concentrations of IL-12 in MLR supernatant in the three groups were (12.6 ±4.4) ng/L, (9.4 ±3.3) ng/L and (8.5 ±3.7) ng/L, and the concentrations of IFNγ were (32. 3 + 9. 2 ) ng/L, ( 27. 4 ± 6. 5) ng/L and (24. 4 ± 7. 4 ) ng/L Both of the values in untreated cases were higher than those of the recovered cases or controls (P < 0. 05 ) , but there were no statistical difference between the recovered and control groups ( P >0. 05 ). The concentration of IL-12 in MLR supernatant correlated positively with the growth rate of lymphocyte (r=0. 529,P <0. 01) and so did the concentration of IFNγ (r = 0. 381, P < 0. 05). Conclusion The function of mDCs to stimulate T lymphocytes proliferation in SAA was enhanced; it might play an important role in the immunopathogenesis of SAA.

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

BACKGROUND:Femoral head avascular necrosis is common in children and elderly. Though, there are many methods can be used for the treatment, it has been inconclusive in the treatment according to the age and stage of the patients. OBJECTIVE:To treat the femoral head avascular necrosis with different methods according to the age and stage, and to retrospectively analyze the fol ow-up results. METHODS:Total y 202 patients (242 hips) with femoral head avascular necrosis were included from October 1998 to October 2008. The patients were divided into groups according to age, included child group (4-14 years old, n=45, 47 hips);youth group (15-45 years old, n=100, 125 hips);and elderly group (46-81 years old, n=57, 70 hips). The patients in the child group were treated with vascularized greater trochanter or iliac periosteal flap displacement, the patients in the youth group were treated with vascularized greater trochanter or iliac periosteal flap displacement, and the patients in the elderly group were treated with hybrid or cementless total hip arthroplasty. RESULTS AND CONCLUSION:Al the 202 patients were fol owed-up for 6 months to 10 years, average 6.3 years. The excellent and good rate of the child group, youth group and elderly group were 87%, 89%and 86%respectively, and al the patients obtained the satisfactory clinical effect;the Harris score of the youth group was increased to (88.1±0.9) points, and the visual analog scale score was decreased to (0.9±0.4) points;the Harris score of the elderly group was increased to (91.5±1.0) points, and the visual analog scale score was decreased to (0.60±0.07) points. The results indicate that vascularized greater trochanter or iliac periosteal flap displacement is suitable for the children and the youths with femoral head avascular necrosis, especial y the patients with the age of 15-45 years in Ficat Ⅱ and Ⅲstage;hybrid or cementless total hip arthroplasty is suitable for the elder patients with femoral head avascular necrosis, as wel as the patients with failure femoral head retention treatment.

ObjectiveTo explore the feasibility,efficacy and cosmetic effect of three-dimensional conformal external beam partial breast irradiation (EB-PBI) after breast-conserving surgery for the selected Chinese early stage breast cancer patients.MethodsFrom June 2003 to December 2010,Forty-four early stage breast cancer patients underwent EB-PBI after breast-conserving surgery.Twenty patients had CT simulation scan in moderate deep inspiration breathing hold,and twenty-four patients in free breathing.EB-PBI was planned and delivered by three-dimensional conformal radiotherapy (3DCRT)with four noncoplanar beams.The prescribed dose was 3.40 Gy per fraction in thirty-nine patients and 3.85 Gy per fraction in five patients,twice per day at an interval of at least six hours,in five consecutive days.Results The number of patients with follow up time of 2,3 and 5 years were 39,31 and 16.Grade 1 acute radiationinduced dermatitis was observed in 17 patients (39％) at three months.Cosmesis was good or excellent in all cases at six months after radiotherapy and in 95％ cases at two years after radiotherapy.The 2-,3-and 5-year local control rates were 100％,99％ and 94％,respectively.The 2-,3-,and 5-year survival rates were all 100％ and no metastases occurred.Conclusions EB-PBI delivered by 3DCRT is feasible for selected Chinese early stage breast cancer patients after breast-conserving surgery.The cosmetic effect,local control rate and long-term survival rate are satisfactory,and acute radiation toxicity is very low.

Objective To explore the expression of antigen activated of macrophages ( MΦ) of bone marrow and its clinical significance in pancytopenia patients with positive bone marrow mononuclear cells (BMMNC)-Coombs test ( immunorelated pancytopenia, IRP) . Methods Sixty-one IRP patients, 10 severe aplastic anemia (SAA) patients and 13 healthy controls were enrolled in this study. The categories of auto-antibodies(IgG, IgM) on BMMNC(CD_(34)~+/CD_(15)~+/GlycoA~+ hematocytes), the quantity (CD_(68)~+/CD_(45)~+)% and expression of antigen activated ( CD_(69) ) of MΦ ( CD_(68)~+ CD_(69)~+/CD_(68)~+ ) % in bone marrow of all cases and controls were measured by fluorescence activated cell sorting( FACS). Results The quantity and expression ratio of activated antigen of bone marrow ( BM ) MΦ in IRP patients [ ( 0. 57 ± 0. 30 ) % and ( 40. 30 ± 18.49)%] were respectively significantly higher than those in SAA [ (0.46 ± 0. 08)% and ( 32. 44 ± 19.37)%] and healthy controls [ (0. 44 ± 0. 69)% and (29.71 ± 11. 67 )% ] ( both P < 0. 05 ). The quantity presented high-positive correlation with the expression ratio of activated antigen of BM MΦ ( r = 0.89, P<0. 01). Patients with IRP were classified into two subgroups according to the quantity of MΦ: Group A (MΦ≥0. 5% , 34 cases) and Group B ( MΦ <0. 5% , 27 cases). Thirty-two cases (94. 12%) were with auto antibody ( IgG) in Group A, while only 2 (7. 41% ) with auto antibody ( IgG) in Group B. There was significant difference in expression ratio of activated antigen of BM MΦ between Group A (49. 19 ± 16. 63) % and Group B (29. 11 ± 14. 30) % ( P < 0. 05 ) , but no difference was found between Group B and the control group (P >0. 05). Total curative rates at 3 and 6 month (47. 06% and 79. 41% ) of Group A were better than those of Group B (22.22% and 51.85%). Thirty-four IRP patients with autoantibody ( IgG) ( + ) were divided into two subgroups according to the quantity of MΦ: high level group ( >0. 75% , 9 cases) and low level group( <0. 75% , 25 cases) , 24 cases (96% ) in MΦ low level group were found auto-antibody (IgG) on one hemotopoietic cell lineage, 1 on two lineages, while 8 (88. 89% ) in MΦ high level group were detected auto-antibody (IgG) on two cell lineages, and 1 on three cell lineages. Expression ratio of activated antigen (56. 12 ± 15. 11) % was much higher in MΦ high level group than that in MΦ low level group (44. 58 ± 18. 16)% (P < 0. 05 ). The count of red blood cell concentration of hemoglobin and platelet in peripheral blood in MΦ high level group were respectively lower than those in MΦ low level group, while the percentage of Ret, the level of total bilirubin and indirect bilirubin, the ratio of erythroid of sternal bone marrow in MΦ high level group were higher than those in MΦ low level group. Conclusion The expression of activated antigen of BM MΦ was enhanced in IRP especially with autoantibody (IgG) , which might be involved in damage process of hemotopoietic cell.

The gene (eg1) encoding for novel endoglucanase 1 was cloned previously from Chinese straw mushroom Volvariella volvacea. EG1 has high thermal stability and optimal pH at neutral and shows great potential in textile and paper industry applications. To improve the expression level of EG1 in Pichia pastoris, the increasing copy number of clone, and its high cell density fermentation in 3.2L fermenter for its high-level expression were investigated in this work. By electro-transformation of pPICZalphaB-egl into GS115EG11 integrated with single copy of eg1 gene, A resistant transformant with 3.8 times higher level expression than GS115EG11 was screened from YPDSZ plate containing 2000 microg/mL of Zeocin. The effect of initial cell density, pH and methanol on its expression and biomass accumulation was evaluated in shaking culture. Optimal EG1 production was observed when initial cell density OD600 was 5.0. EG1 production and biomass accumulation did not seem to vary when cells were induced at different pH values. Both of EG1 and cell density were found to increase with higher methanol concentrations, reaching 62.48 IU/mL and 31.7 (OD600) respectively after 120 h induction with 2.0% (V/V) methanol compared to 30.24 IU/mL and 17.79 (OD60) with 0.25% methanol induction. EG1 expression was further increased by 6.4 times higher than shaking culture after 95.5 hours induction with methanol in fed-batch fermentation, so totally 34 times higher than that for GS115EG11 was achieved by screening of high Zeocin resistant clone and high cell density fermentation. The production of EG1 with 543.36IU/mL CMC activity and 8.80mg/mL protein expression was obtained in Pichia pastoris.
Cellulase ; biosynthesis ; genetics ; Cloning, Molecular ; Culture Media ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Volvariella ; enzymology ; genetics

Objective To explore the inhabitants' knowledge about Keshan disease(KD) in endemic area in Heilongjiang Province and analyze its influencing factors. Methods Yongjin Village, a national KD monitoring site in Fanrong Township of Fuyu County in Heilongjiang Province,was selected as an investigating spot. Specially designed questionnaire was applied in the investigation, which included three aspects, demographical information (gender, age, educational level, etc.), KD-related knowledge survey (KD prevention measures including a healthy diet supplementing selenium, preventing microbial infection, inducers such as climate change, fatigue, cold, mental shock, overeating) and KD controlling measures (take prescribed medicine, dietetic regulation and regular exercise, etc.) Results Seventy-senven point six percent(272/352) knew the disease by the name in the endemic area, while only 29.7% (81/272)and 30.8% (84/272)were aware the preventive measures and inducers , respectively. The awareness rates of KD preventive measures and inducers among the KD patients were 6/17 and 9/17 respectively. The influencing factors of the awareness rate of KD name were gender, vocation and the state of education (χ2 value were 9.838,9.878,12.462, all P < 0.05). However, factors influencing the awareness rates of preventive measures were gender and the state of education(χ2 value were 7.400,20.251, both P < 0.05). Conclusions The awareness rates of KD preventive measures and inducers axe low in the endemic area. The major factors influencing the awareness rate of KD are gender and the educational degree.