Object To study the feasibility of suspension culture of protocorm in Dendrobium candidum Wall ex Lindl and effect of inoculum and medium volume on the growth of protocorm Methods Effect of four basic media MS, 1/2 MS, 67 V, and B 5, inoculum and medium volume on the growth of protocorm were studied by completely random experimental design and orthogonal test design Results The growth of D candidum protocorms in liquid medium was markedly better than that in solid medium (P0 05), B 5 was much better than 1/2 MS (P

20 first mandibular premolars were randomly divided into 2 groups(n =10).The teeth in experimental group were treated by fi-ber main post in combination with auxiliary pile,in control group by single fiber main post,and then were restored by metal crown.They were fixed in universal testing machine.The fracture load(N)of experimental and control group was (846.50 ±40.33)and (437.90 ±41.15) respectively(P <0.01).

Objective To study the expressions of hypoxia inducible factor (HIF)-1 and angiopoietin (Ang)-2 in repeated gastrointestinal bleeding due to vascular malformation, and the efficacy of treatment with thalidomide. Methods Twenty-five patients with repeated gastrointestinal bleeding due to vascular malformation confirmed by capsule endoscopy or enteroscopy were collected and 18 subjects without severe diseases were served as controls. Ten patients with gastrointestinal vascular malformation, who received 25 mg of thalidomide 4 times daily for 4 months and were followed up for at least one year, were also enrolled. The serum samples from all participauts were detected for expressions of HIF-1 and Ang-2 using enzyme-linked immunosorbent assay (ELISA).The expressions of HIF-1 and Ang-2 were compared between angiodysplasia group and control group.The expressions of HIF-1 and Ang-2 were comparatively evaluated before and after treatment with thalidomide in treatment group. Results The expressions of HIF-1 and Ang-2 in vascular malformation group [( 113. 84 ± 26. 66 ) ng/ml and ( 652. 11 ± 140. 39) ng/ml, respectively] were significantly higher than that of control group [(43.28±17.30) ng/ml and (265.60±53.88) ng/ml,respectively, P=0. 000]. The expression of HIF-1 was positively associated with that of Ang-2. (r=0. 700, P= 0. 000). There was no difference in expressions of HIF-1 and Ang-2 before and after treatment with thalidomide (P=0. 498 and =0. 136, respectively). However, significant reduction of Ang-2 [(113. 80±73. 60) ng/ml(P=0. 003)] was found in 8 effectively treated patients after thalidomide treatment. Conclusions HIF-1 and Ang-2 might play an important role in the formation of vascular malformation. The extent of Ang-2 reduction after thalidomide treatment may be helpful in evaluating its efficacy or prognosis.

ObjectiveTo study the pathogcncsis of gastrointestinal vascular malformation (GIVM) and the potential mechanism of thalidomide in the treatment of gastrointestinal bleeding due to GIVM.Methods We collected the surgical intestinal specimens from 10 patients who suffered from massive hemorrhage of gastrointestinal tract owning to GIVM and the normal intestinal mucosa around the lesions,as well as normal intestinal mucosa from healthy subjects.Immunohistochemical(IHC) staining was carried out to investigate the differences of angiopoietin 2 ( Ang2 ),Notch1 and delta like ligand 4 (Dll4) in the above three intestinal mucosa to find the relationship with the pathogenesis of GIVM. Human umbilical vein endothelial cells(HUVECs) were cultured with 0,25,50,100 and 200 mg/L thalidomide for 24 or 48 hours to observe their mRNA and protein expressions of Ang2,Notch1,Dll4 by real-time PCR and Western blot.ResultsBy IHC staining,more expressions of Ang2,Notch1 and Dll4 in the lesions were detected than those in the normal intestinal mucosa around the lesions and the normal intestinal mucosa in healthy people.The expressions of Ang2,Notch1 and Dll4 were significantly correlated (P =0.016,r =0.732),and the expressions of Notch1 and Dll4 were absolutely correlated ( P =0.000,r =1.000).Real-time PCR and Western blot showed that thalidomide could down-regulate the expressions of them,which were in a concentration-dependent manner.ConclusionAng2,Notch1 and Dll4 may correlate with the pathogenesis of GIVM,while thalidomide can concentration-dependently down-regulate the expression of Ang2,Notch1 and Dll4,which may be one of the mechanism that thalidomide play a therapeutic role in GIVM.

Objective To investigate the inhibitory effect of thalidomide on angiodysplasia.Methods Excisional intestinal specimens were collected and immunohistochemical examination was carried out.The human umbilical vein endothelial cells were cultured in vitro to exponential phase of growth,divided into six groups and synchronized for 24 hours.They were then stimulated with thalidomide (40-100 μg/ml) for 72 hours.MTT assay was used to assess cellular proliferation.ELISA,real-time quantitative PCB and western blot were applied to detect the expression of VEGF/HIF-1α of human umbilical vein endothelial cells (HUVEC).Results Immunohistochemical analysis of intestinal pathological specimens demonstrated higher expression of VEGF.ELISA showed that the expression of VEGF under hypoxia was obviously higher than that under normoxia [ ( 1199.3 ± 61.4) ng/L vs ( 864.7 ± 41.2 ) ng/L,P < 0.05 ].Real-time quantitative PCR and Western blot discovered that thalidomide inhibited the expression of VEGF/ HIF-1α of HUVEC (P < 0.05).The effect of thalidomide was dose-dependent.Conclusions Thalidomide can suppress the expression of HIF-1α and VEGF in HUVEC in vitro and then inhibit angiodysplasia,which may play a significant role in stopping the rebleeding in patients with recurrent gastrointestinal bleeding.

Objective To compare the efficacy between the two-cuff swan neck catheter and the Tenckhoff catheter in continuous ambulatory peritoneal dialysis (CAPD) patients prospectively. Methods One hundred and ten patients with end-stage renal disease (ESRD) were selected as candidates, who received catheter implantation and CAPD therapy for the first time. Patients were divided into group A (swan neck catheter group) and group B (Tenckhoff catheter group), 55 patients for each group. Catheters of beth groups had a straight end and were implanted by routine surgical procedure. One-year follow-up was performed and information was recorded such as complications, survival time, quit of dialysis, death, etc. Survival analysis was carried out by Kaplan-Meier method and Log-Rank tests. Results At the end of follow-up, 17 patients died, 3 received renal transplantation, 8 were transferred to hemodialysis, 3 went to other hospitals, and 79 patients (71.8%) remained in our department for CAPD. Twenty-six patients of both groups had peritonitis with a total of 35 occurrences. The total incidence of peritonitis was 0.32 times/patient year, with the detailed figure of 0.35 times/patient year for group A and 0.29 times/patient year for group B respectively (P0.05). The time interval between the catheter implanting and the onset of peritonitis was (30±29) weeks and (29±24) weeks for group A and group B respectively (P0.05). The risk of developing peritonitis in both groups was 26.97% within 1 year. Tunnel infection occurred in 2 patients and exit-site infections in 9 patients of two groups. The incidence of tunnel plus exit-site infections was 0.1 times/patient year. Incidence of tunnel infection and the exit-site infection for group A was lower than that of group B (0 vs 0.036 times/patient year and 0.06 times/patient year vs 0.11 times/patient year respectively). However, the difference was not significant (P0.05). Mechanical complications of catheter (catheter migration, omcntum wrapping, leakage of peritoneal dialysates, slip out of outer cuff), incidence of inguinal hernia and bellyache between two groups were not significantly different (P0.05). There were 4 cases of catheter drawing in each group. Both two groups had the same 12-month technical survival rate as 92.73%. Of 17 dead cases, 7 were in group A and 10 in group B (P0.05). The main death causes were cardiocerebral events (47.1%) and infections (23.5%). The 12-month survival rate was 86.34% for group A and 80.68% for group B (P0.05). Conclusions There are no significant differences of infection, mechanical complications, technical survival rate and patients' survival rate between two groups. The efficacy of swan-neck catheter is similar to Tenckhoff catheter in CAPD patients.

Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P

Objective To explore humam cytomegalovirus(HCMV) encoded microRNAs during latent infection in order to help study HCMV virology and latent infection mechanisms.Methods A model of HCMV latent infection via THP-1 cells infected with HCMV was constructed.Deep-sequencing was performed using high-resolution Solexa sequencing platform.The secondary structure of the newly sequenced miRNA was predicted by RNAFold bioinformatics software. Results HCMV encoded various miRNAs during latent infection, including miR-US25-2-3p, miR-US25-2-5p, miR-UL112, miR-US25-1, miR-UL22A and PC-5p-148467 with a predicted length of 25 bp, named hcmv-miR-US33as-5p.Conclusion HCMV can express many types of miRNAs during latent infection.

Objective:To investigate the effects of rituximab on lymphocytes and immunoglobulin in the treatment of focal segmental glomerulosclerosis (FSGS) and minimal change disease (MCD).Methods:The subjects were FSGS and MCD patients admitted to Ruijin Hospital affiliated to Shanghai Jiaotong University on July 1, 2014 and July 1, 2019. All the enrolled patients were confirmed by clinical examination and renal biopsy, and received rituximab treatment (4 infusions of 375 mg/m 2 with the interval of 7-14 d). The levels of immunoglobulin IgA, IgG, IgM, and lymphocytes of CD19 +, CD20 +, CD3 +, CD3 +CD4 +, CD3 +CD8 + and natural killer cells (CD56 +CD16 +) were compared between baseline and the third month, the sixth month, the ninth month and the twelfth month after treatment. Results:Ninety-six patients with FSGS or MCD were enrolled in this study. The midian age was 28 years old (14-77 years old). The ratio of men to woman was 1.8∶1. There were 65 cases of MCD and 31 cases of FSGS. After rituximab treatment, the 24 h-proteinuria was significantly lower than that before treatment, and the serum albumin level was increased (both P<0.05). After rituximab treatment of 3 months, 6 months, 9 months and 12 months, CD19 + and CD20 + lymphocyte counts were significantly decreased (all P<0.01), and gradually recovered after 6 months. Compared with baseline, at 3, 6, 9, 12 months after rituximab treatment, the level of blood IgG was significantly increased ( P=0.004,<0.001,<0.001,<0.001, respectively), and the level of blood IgM was significantly decreased ( P<0.001, =0.008, =0.005,<0.001, respectively) but the median level still within the normal range (400-3 450 mg/L). The level of blood IgA was not significantly changed (all P<0.05). T lymphocytes (CD3 +, CD3 +CD4 + and CD3 +CD8 +) and natural killer cells (CD56 +CD16 +) showed no significant difference from baseline (all P>0.05). Conclusions:Rituximab can effectively eliminate CD19 + and CD20 + lymphocytes, and has little influence on peripheral blood lymphocyte count and immunoglobulin level except CD19 + and CD20 + lymphocytes. The standard administration of rituximab is safe for patients with FSGS and MCD.

To obtain a recombinant human plasminogen (hPLG) with potential anti-platelet aggregation activity, we cloned the cDNA coding Pro544 to Asn791 of hPLG, a kringle-deficit derivative (hPLG-deltaK). The Pro559 in activation loop was then mutated into Asp559 to provide Arg-Gly-Asp (RGD) motif. The constructed pPICZalphaA-RGD-HPLG-deltaK plasmid was expressed in yeast Pichia pastoris GS115, which produced RGD-hPLG-deltaK about 0.160 g/L broth. After affinity chromatography, the purity of the recombinant protein reached above 90%. Western blotting test confirmed that it retained the immunological reaction capability as human PLG. Its urokinase activation rate in 24 hours and its fibrinolytic activity made no deference against native hPLG-deltaK (P=0.630, n=5). Importantly, after activation by urokinase, RGD-hPLG-deltaK showed a significantly higher platelet aggregation inhibition rate (Ri) (21.8% +/- 1.57%) than hPLG-deltaK (3.8% +/- 0.33%) (P=0.000, n=5). These results proved that we constructed an hPLG mutant with anti-platelet aggregation activity, which made a foundation for developing innovative thrombolytic drugs with multifunction.
Humans ; Kringles ; genetics ; Oligopeptides ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Plasminogen ; biosynthesis ; genetics ; Platelet Aggregation Inhibitors ; isolation & purification ; pharmacology ; Point Mutation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; pharmacology