Background The pathogenesis of age-related cataract is associated with the apoptosis of lens epithelial cells (LECs) caused by oxidative stress.Previous studies showed that intracellular focal adhesion kinase (FAK) pathway can be activated by H2O2 in vitro,which induced apoptosis of cells.To investigate the effect of oxidative on FAK expression in LECs is one of important studies in the prevention of age-related cataract.Objective This study was to investigate the expression and function of FAK in human LECs treated by H2O2.Methods Human LECs strain (HLECs-B3) were cultured in vitro in the low glucose DMEM with 10％ fetal bovine serum.Different concentrations (0,30,50,70,100,300,500,700,1000 μmol/L) of H2O2 were added into the culture medium for 24 hours.The survival rate of the cells was detected by Cell Counting Kit-8 (CCK-8) assay.Cell morphology as well as the expression and distribution of FAK in the cells were observed by immunofluorescent staining under the laser confocal microscope.Apoptosis was observed by hoechst33258 staining,and Western blot assay was used to quantitatively detect the expression and phosphorylation of FAK.Results The survival rate of the cells was (1.00±0.03) ％,(1.24±0.03)％,(1.36±0.24) ％,(0.93±0.02)％,(1.75±0.19)％,(1.37±0.18) ％,(0.64±0.01)％,(0.59±0.11)％,(0.14±0.05)％ in 0,30,50,70,100,300,500,700,1000 μmol/L H2O2 groups,with a significant difference among them (F =95.30,P =0.00).The survival rates of the cells in the below 300 μmol/L H2O2 groups were significantly higher than those in the 0 μmol/L H2O2 group,and survival rates of the cells in the above 500 μmol/L H2O2 groups were significantly lower than those in the 0 μmol/L H2O2 group(all at P＜0.05).After H2 O2 treatment for 24 hours,HLECs-B3 cells transformed from polygon shape to spindle shape and extended pseudopodiums,meanwhile the green fluorescence for FAK exhibited in the cytoplasm.Cell apoptosis was found in the 1000 μ mol/L H2O2 group.Western blot assay revealed that the expressing levels (grey scale) were significantly different among the various groups (F=28.08,P=0.00),and FAK expressing levels in the below 300 μmol/L H2O2 groups were significantly higher than those of the 0 μmol/L H2O2 group; while the expressing levels in the above 500 μmol/L H2O2 groups were lower than those of the control 0 μmol/L H202 group (all at P＜0.05).After treated by different concentrations of H2O2,the phosphorylation level of intracellular FAK (p-FAK) was significantly higher in 3 hours group than that in 30 minutes group (all at P＜0.05).Conclusions H2 O2 can affect the survival,proliferation and morphology of human LECs by activating the intracellular FAK pathway,indicating that FAK may play roles in the regulation process of cell biological behavior.

The study evaluates the lipolysis rate and extent of type Ⅲ lipid formulations using testosterone undecanoate as a model drug after digestion with in vitro lipolysis model, and studies the digestive regularity with optical microscope and electrical conductivity. The results showed that for testosterone undecanoate type Ⅲ lipid formulations with castor oil as oil phase and Transcutol HP as latent solvent, the lipolysis rate and extent were increased with the increase of oil phase proportion and were decreased with excessive proportion of surfactant, in which can see liquid crystal phase during lipolysis process. The lipolysis rate of type ⅢB lipid preparations with different surfactant were ordered as Labrasol > Tween 80 > Cremophor EL, but the rate of type ⅢA is different in quick digestion phase and slow digestion phase. The lipolysis extent of type Ⅲ lipid formulations with different surfactant were ordered as Cremophor EL > Tween 80 > Labrasol. These may be related to the digestive effect of pancreatic lipase on different surfactants. This study implied that the lipolysis rate and extent of type Ⅲ lipid formulations are greatly influenced by the proportion of oil phase and surfactant, and the surfactant structure. These factors will affect the in vivo digestion and should be taken into account when screening type Ⅲ lipid formulations.

Objective:To study the abnormal suppressive capacity of regulatory T cells ( Tregs) to effector cells,Th17 cells and Th9 cells in patients with mild to moderate asthma attack.Methods: Recruited asthmatic patients (n=30) and healthy control (n=30),collected peripheral venous blood from healthy control and asthmatic patients in mild to moderate attack stage respectively,and then isolated CD4+CD25+CD127-/low Tregs and CD4+CD25-T effect cells with immunomagnetic beads method ( purity was above 90%).The effect cells were cultured with PHA stimulation with or without Tregs.Measured the proliferation (3H-Thymidine), expression of RORC and PU.1 ( RT-PCR) genes,production of IL-17 and IL-9 ( LUMNEX) in effect cells with or without Tregs inter-vention.Analyzed the abnormality of Th17 and Th9 cells and the supressive capacity of Tregs on Th17 and Th9 cells in patients with asthma attack,comparing with control group.Results:Tregs of asthmatic patients could suppress the proliferation of effector cells,but less effectively than healthy control (P=0.03).In asthmatic patients,RORC expression and IL-17 production increased (P<0.05), and the suppressive capacity of Tregs to RORC expression decreased, when compared with healthy group ( P<0.05 ) .In asthmatic patients,IL-9 production of effector cells increased significantly (P<0.05),but PU.1 expression was higher than healthy group only with intervention of Tregs ( P=0.04 ) .And the suppressive capacity of Tregs to the expression of PU.1 and production of IL-9 in patients with asthma attack did not decrease (P>0.05).Conclusion:The suppressive capacity and amount of Tregs decreased,but the specific gene expression and cytokines production of Th17 cells and Th9 cells were upregulated in patients with mild to moderate asthma attack.This deficiency for the suppressive capacity of Tregs to Th17 but not Th9 cells may indicate some pathogenesis of asthma devel-opment.

Objective To explore the effect of reconstructing unilateral cleft lip by changing the arc-shaped incision, combined with the 3D reconstruction of upper lip muscles.Methods Twenty unilateral cleft lip patients were treated by using a new surgical operation, the 3D reconstruction of upper lip muscle, to restore normal anatomy and stress of the mucous membrane, muscle and skin.Operation scar was designed for straight line, located on the philtral ridges of the contour line;phitrum and philtral ridges were rebuilt, and postoperative scar reduced.Results A lot of 20 patients had no local infection, hemorrhage, complex crack, and were stage I incision healing.Followed up for 1-8 months postoperatively, the patient's lip bow line continuity was good, with symmetrical shape and good phitrum and philtral ridges;scar was hidden on the philtral ridges of the contour line, and no obvious upper lip scar contracture found through the follow-up period.Conclusions This improved method is simple in the incision design, and less scar hidden on the philtral ridges of the contour line after operation, which can maximize the recovery of the appearance of nose and upper lip with satisfactory effect.It is a feasible improvement method of repairing unilateral cleft lip.

OBJECTIVE:To virtually screen lignanoid compounds with inhibitory effect of histone deacetylase(HDAC)by vir-tual screening method. METHODS:Using“lignanoid”as keyword,requiring CNKI,VIP,PubMed and other database,lignanoid compounds were collected as ligand to establish ligand base, histone deacetylase receptor HDAC2 (PDB code:4LXZ) and HDAC8 (PDB code:1T69) were selected from PDB database,and then ligands and 3D active site of receptors were docked by SYBYL-X 2.0 software. The affinity of receptors to ligand was reflected by total score. RESULTS:345 lignanoid compounds, 4LXZ and 1T69 primary ligand were used to establish ligand base which included 347 ligands. Ligands No.275,271,110,200, 056,258,181,129,037,270,187 were demonstrated good affinity with receptors HDAC2 and HDAC8. Ligands and receptors residue were docked via hydrogen bond. CONCLUSIONS:Lignanoid compounds have inhibitory effect on HDAC;virtual screen-ing method is effective in natural product activity prediction,which can provide quick access to and theoretical guidance for new pharmacological studies of lignanoid compounds.

Abstract: Objective To establish the duplex TaqMan RT-PCR method for detection of Entamoeba histolytica and Giardia lamblia in fecal samples. Methods Primer pairs and probes for Entamoeba histolytica and Giardia lamblia were designed and duplex TaqMan RT-PCR amplification system was constructed. PCR products were inserted into the pUC57 plasmid, and the lower limit of detection of the method was determined. Clinical stool samples were tested in order to evaluated the efficacy of the method. Results The detection limits of duplex TaqMan RT-PCR were 31.6 copies/μL for Entamoeba histolytica and 32 copies/μL for Giardia lamblia, respectively. Of the total of 212 clinical stool samples tested, all 3 samples with E. histolytica-positive patients by microscopy were positive by PCR, while 1 from 209 samples with E. histolytica-negative patients by microscopy were positive by PCR, and the remaining samples were negative. For Giardia lamblia, all 8 samples positive by microscopy were positive by PCR, and 1 from 204 sample with a microscopy-negative patient was positive by PCR, and the remaining samples were negative. The amplification product sequencing and blast analysis were used to confirm that the amplified sequence in the specimen of a patient with negative microscopy but positive PCR belongs to the targeted pathogen, supported by clinical symptoms and laboratory test results. PCR results for other diarrhea-causing pathogens were negative, indicating no cross-reactivity. Conclusions The dual TaqMan RT-PCR method developed in this study can not only detect microscopy-positive samples of Entamoeba histolytica and Giardia lamblia but also can detect samples that were missed by microscopy, with higher sensitivity than the microscopy method. Further, this detection method does not cross-react with other diarrhea-causing pathogens, including cross-react with other diarrhea-causing pathogens including Iodamoeba butschlii, Blastocystis hominis, Plesiomonas, Aeromonas, Salmonella, Shigella, Sphaerozoum fuscum, and Entamoeba hartmani, thus has a good specificity.

Adolescent ; Child ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Killer Cells, Natural ; pathology ; Leukocyte Common Antigens ; analysis ; Lymphoma, T-Cell ; genetics ; immunology ; Male ; Microtubule-Associated Proteins ; genetics

Objective: To analyze the stress difference of alveolar bone around the abutment and alveolar ridge of edentulous of three different kinds of root-attachment-supported overdenture under different load conditions, in order to provide a reference for the choice of clinical root attachment. Methods : The occlusal force of overdenture was simulated by electrical resistance strain measurement in vitro. The stress of the alveolar bone, the central part of the mandibular arch and the first molar correspond to the alveolar ridge were measured. The stress difference of 3 kinds of attachment overdenture under different loading conditions were compared and analyzed. Results: Under the same loading condition, all three kinds of overdentures had a certain degree of slip of the denture (magnetic attachment denture) or rotation (3 kinds of attachment dentures). The abutment neck in different parts of the dental arch and alveolar bone, anterior free end edentulous alveolar ridge stress distribution was significantly different. Conclusion ERA attachment overdenture was the most preferable, followed by the magnetic attachment overdenture. Suitable attachment should be selected based on specific clinical cases.

Objective: To evaluate the prognostic value of kinetic changes in minimal residual disease (MRD) status, as well as its relationship with risk stratification, therapeutic response and treatment in patients with newly-diagnosed multiple myeloma (MM) . Methods: A total of 135 patients with newly-diagnosed MM were screened, and 105 patients who achieved VGPR or more as the best responses were included into this study. The MRD status was determined by multiparameter flow cytometry (MFC) at multiple intervals after two cycles of treatment until clinical relapse, death, or last follow-up. The statistical methods included Kaplan-Meier analysis, Cox regression, etc. Results: ①In all 135 patients, 57.8% (78/135) patients achieved MRD negativity (MRD(-)) after treatment. In 105 patients who achieved VGPR and thus included in this study, the MRD(-) rate was 72.4% (76/105) , with a median interval of 3 months from starting treatment to achievement of MRD(-) status. ②The 2-year PFS rate of patients with MRD(-) status was significantly higher than that of MRD(+) status (62.2% vs 41.3%, P=0.001) , while MRD persistence (MRD(+)) was an independent factor for poor prognosis (multivariate analysis for PFS: P=0.044, HR=3.039, 95%CI 1.029-8.974) . ③Loss of MRD(-) status (i.e., MRD reappearance) showed inferior outcomes compared with MRD sustained negative ones, the PFS was 18 months versus not reach (P<0.001) and the OS was not reach for both (P=0.002) . ④The 2-year PFS and OS rates of patients with duration of MRD(-)status≥12 months were significantly higher than those of the control group (PFS: 77.7% vs 36.7%, P<0.001; OS: 96.4% vs 57.9%, P<0.001 respectively) . Duration of MRD(-) status was associated with a marked reduction in risk of relapse or death (univariate analysis for PFS: P<0.001, HR=0.865, 95%CI 0.815-0.918; for OS: P=0.001, HR=0.850, 95%CI 0.741-0.915 respectively) . ⑤Moreover, even in patients carrying high-risk cytogenetic abnormalities (CA) or ineligible for ASCT, MRD negativity remained its prognostic value to predict PFS (high-risk CA medianPFS: not reach vs 19 months, P=0.006; ineligible for ASCT medianPFS: not reach vs 25 months, P=0.052 respectively) . ⑥Last, treatment with the bortezomib-based regimens contributed to prolonged MRD(-) duration (median MRD(-) duratio: 25 months vs 10 months, P=0.034) . Conclusion: Our findings supported MRD(+) status as an independent poor prognostic factor in MM patients, which implicated that duration of MRD(-) status also played a significant role in evaluation of prognosis, while loss of MRD(-)status might serve as an early biomarker for relapse. Therefore, monitoring of MRD kinetics might more precisely predict prognosis, as well as guide treatment decision, especially for when to start retreatment in relapsed patients.
Bortezomib/therapeutic use* ; Humans ; Multiple Myeloma/therapy* ; Neoplasm Recurrence, Local ; Neoplasm, Residual/diagnosis* ; Prognosis ; Risk Assessment ; Treatment Outcome