Endocarditis, Bacterial ; Female ; Humans ; Hypertension, Pregnancy-Induced ; Pregnancy

Objective To investigate the possible mechanisms of enhancing angiogenesis and arteriogenesis through the oberservation of the effect of gene transfer of angiopoietin-1 (Ang1) on its receptor Tie2 in rat models of acute myocardial infarction. Methods Myocardial infarction was induced in rats by left anterior descending artery ligation. Naked plasmid DNA encoding human angiopoietin-1 (phAng1) was delivered into the ischemic area (group A) by intramyocardial injection. On day 3, 7, 14 and 28 after the injection, the mRNA expression of Tie2 and its changes with time were determined by RT-PCR. The number of vessels and arterioles was examined by immunohistochemistry. The collagen was evaluated by Masson staining. Results RT-PCR showed that mRNA expression levels of Tie2 in group A were significantly higher than those in the control group, reached the highest level on day 7 post-injection, and gradually declined to normal level 28 days later. On day 7, 14 and 28, the vessel count showed the number of blood vessels (angiogenesis and arteriogenesis) in group A was greater than that of the control group at the same timepoint and the infracted myocardium in group A was significantly less than that of the control group. Conclusion Gene transfer of phAng1 enhances angiogenesis and arteriogenesis in acute myocardial infarction and reduces the infraction area probably by upregulating the expressions of Tie2 receptor.

Objective Observed the mobilization effects、 the safety and feasibility of autologous circulating blood stem cell by G-CSF in acute myocardial infarction (AMI). Methods 45 patients with AMI were randomly allocated to receive either inclusive type Granulocyte Colony-Stimulating Factor (G-CSF), to mobilize the stem cell. The patients received the dose of G-CSF 300?g-600?g/day, by hypodermic injection, and the duration of applying G-CSF was 5 days. In the process of the mobilization of the circulating blood stem cell, prior to applying G-CSF and on the 3rd、4th、5th、6th、7th after applying G-CSF, the white blood cell (WBC) and CD34+ cell count in the circulating blood should be observed; and the following side effects also should be paid attention to, such as: bone pain, tetter, fever, gastrointestinal effects( nausea 、vomit、 coprostatis ),deteriorated of angina or heart failure and some rare effects(spontaneous spleen rupture, severe purulent infection, hypercoagulable state, autologous immune diseases). Results Prior to applying G-CSF and the 3rd、4th、5th、6th、7th after applying G-CSF, the counts of WBC were (8.42?2.59)?10 9/L、(31.28?8.34)?10 9/L、(35.24?9.38)?10 9/L、(37.03?13.07)?10 9/L、(35.34?14.68)?10 9/L、(20.35?9.22)?10 9/L;the counts of CD34+ cell were (14.89?11.46)?10 6、(67.78?50.88)?10 6、(124.79?136.13)?10 6、(208.92?206.97)?10 6、(206.10?184.57)?10 6、(66.63?56.56)?10 6;The peak of curve that WBC and CD34 + cell count changed with applying days was on the 5th .The count of CD34 + cell in the circulating blood was positive referent with the count of WBC in the circulating blood(r=0.835);was not reference with age、 sex、body weight、and the onset time of AMI. There were total 17 complications during the mobilization of circulating blood stem cell. The incidence of complications during mobilization is 37.8%(17/45), including bong pain being 15.6%(7/45)、fever being 6.7 %(3/45)、pale being 4.4 %(2/45)、tetter being 4.4 %(2/45)、deterioration of heart failure being 4.4 %(2/45),spleen thrombosis being 2.2 %(1/45).No death happened. Conclusion : In patients with AMI, the mobilized peak of WBC and CD34 + cell counts changed with applying days was at the 5th, and the count of CD34 + cell in the circulating blood was positive referent with the count of WBC in the circulating blood(r=0.940),was negative referent with body weight of patients(r=-0.398). And mobilization of autologous circulating blood stem cell was feasible and safe.

Objective To explore the value of somatostatin receptor scintigraphy(SRS) in evaluating the immune activity of orbital inflammatory pseudotumor associated with systemic vasculitis.Methods Twenty-five patients with orbital inflammatory pseudotumor associated with systemic vasculitis (10 males,15 females,average age:(51.2± 14.2) years) underwent SRS.The uptake ratio (UR) of orbital inflammatory pseudotumor was obtained.(1) Patients were divided into group A (with immune activity) and group B (without immune activity) according to Birmingham vasculitis activity score (BVAS).The difference of UR between the 2 groups was compared by two-sample t test.The difference of UR before and after treatment in 12 patients was also compared.(2) Based on the results by BVAS,ROC curve was used to obtain the cut-off value of UR,as well as the diagnostic efficiency and Youden index.The consistency between SRS and BVAS was calculated.(3)Patients were divided into two groups according to the cut-off value of UR and the prognosis difference between them was compared by Fisher exact test.(4)The expression of SSTR2 and SSTR5 was observed by immunohistochemistry.Results (1) UR in group A was significantly higher than that in group B (2.09±0.44 vs 1.32±0.46,t =5.94,P＜0.01).After glucocorticoids treatment,the UR in group A reduced significantly (t=4.07,P＜0.01),but not in group B (t=1.76,P＞0.05).(2)ROC curve analysis identified UR cut-off value as 1.66,with the sensitivity of 87.5％,specificity of 95.7％,positive predictive value of 95.2％,negative predictive value of 88.0％,accuracy of 91.3％ and Youden index of 83.2％.The consistency between SRS and BVAS was strong (Kappa =0.840).(3) The prognosis was significantly different between patients with UR≥ 1.66 and UR＜1.66 (P＜0.05).(4) The immunohistochemical results revealed high expression of SSTR2 and SSTR5 in inflammatory cells in patients with immune activity.Conclusion SRS has potential value in evaluating the immune activity of orbital inflammatory pseudotumor associated with systemic vasculitis.

BACKGROUND:Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰmaybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately.
OBJECTIVE:To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells.
METHODS:Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in celllines.
RESULTS AND CONCLUSION:The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successful y constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells.

Objective:To observe the clinical efficacy of Dushen Decoction and Zhenwu Decoction in the treatment of chronic congestive heart failure (CCHF) of Yang deficiency and blood stasis type. Methods:From March 2017 to December 2019, 120 patients with CCHF of Yang-deficiency and blood stasis type admitted to Shanghai Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine were selected, and they were divided into study group and control group according to the randomized number table method, with 60 in each group. The control group was given western medicine of conventional treatment, and the study group was combined with Dushen Decoction and Zhenwu Decoction on the basis of the control group. Both groups were treated for 1 month. Before and after treatment, TCM syndrome scores were performed, serum TNF-α, IL-17 and CRP were detected by ELISA, left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) were detected by color doppler echocardiography the adverse reactions during the treatment were recorded and the clinical efficacy was evaluated. Results:The total effective rate was 91.7% (55/60) in the study group and 71.7% (43/60) in the control group, with significant difference between the two groups ( χ2=8.015, P=0.005). After treatment, the scores of palpitation, dyspnea, chilly limbs, dull tongue and edema in the study group were significantly lower than those in the control group ( t=13.953, 13.915, 30.945, 32.339, 20.403, P<0.001). After treatment, LVEF [(56.28 ± 4.34)% vs. (42.47 ± 4.56)%, t=16.993] in the study group was significantly higher than that of the control group ( P<0.01); LVEDD [(44.32 ± 6.23) mm vs. (53.81 ± 5.19) mm, t=9.066] and LVESD [(31.28 ± 4.62) mm vs. (37.51 ± 4.73) mm, t=7.299] in the study group were significantly lower than those in the control group ( P<0.01). After treatment, the serum levels of TNF-α, IL-17 and CRP in the study group were significantly lower than those in the control group ( t=12.644, 15.975 and 14.379, P<0.001). The incidence of adverse reactions was 6.7% (4/60) in the control group and 8.3% (5/60) in the study group, and there was no significant difference between the two groups ( χ2=0.120, P=0.729). Conclusion:Dushen Decoction and Zhenwu Decoction in the treatment of CCHF can improve the clinical symptoms, improve the cardiac function, reduce inflammatory factors, improve the treatment efficiency with good safety.

To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Genetic Vectors ; Interleukin-18 ; biosynthesis ; Lactococcus lactis ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Objective To investigate the effect of 5-azacytidine(5-AZA) on apoptosis of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs were isolated from bone marrow of mouse tibia and femur; the expression of MSC specific markers CD44 and CD90 in BMSCs was measured by immunofluorescence staining;BMSCs were cultured in vitro in the medium supplemented with 0,10 and 20 μmol/L 5-AZA for 48 hours.Cell apoptosis was measured with fluorescent labeled inhibitor of caspases (FLICA) apoptosis kit and 4',6-diamidino-2-phenylindole (DAPI) staining ;the expression of apoptosis-related proteins Annexin V and Caspase-3 in the treated BMSCs was detected by Western blot.Results In this study,BMSCs positively expressed MSC specific markers CD44 and CD90.DAPI staining and Caspase-3 staining both showed that 10 and 20 μmol/L 5-AZA markedly increased apoptotic rate of BMSCs;the apoptosis-positive rate in DAPI staining was (21.086 ± 2.601) ％,(34.467 ± 3.724) ％ and (46.512 ± 3.864) ％,the apoptosis-positive rate in Caspase-3 staining was (5.354 ± 0.735)％,(15.462 ± 2.385)％ and (28.190 ± 4.190)％ in the controls,10 and 20 μmol/L 5-AZA groups,and there were significant differences among the control group and 5-AZA treated groups(all P ＜0.01).Western blot assay showed that Annexin V and Caspase-3 were both markedly upregulated in 5-AZA treated cells;the relative level of Annexin V expression was(26.612 ±2.184)％,(42.873 ±4.313)％ and (50.056 ± 4.457) ％,the relative level of Caspase-3 expression was (19.231 ± 2.683) ％,(38.618 ± 5.385) ％ and(91.235 ± 7.116)％ in the controls,10 and 20 μmol/L 5-AZA groups,and there were significant differences among the control group and 5-AZA treated groups (all P ＜ 0.01).Conclusion The commonly used doses of 5-AZA can induce apoptosis of BMSCs.

Objective To elucidate the effect of bioimpedance ratio in the calf (calf-RBI) for volume evaluation on hypertension in hemodialysis (HD) patients. Methods Bioimpedance in the right calf was measured by bioimpedance spectroscopy (BIS). As an index of volume status, calf-RBI was calculated as follows: calf-RBI =impedance at 200 kHz / impedance at 5 kHz. The range of age-stratified 1SD from mean calf-RBI in the healthy control was assumed as the target range for the corresponding HD patients. The dry body weight (DBW) was stepwise decreased under the guidance of calf-RBI. The changes of calf-RBI, blood pressure and antihypertensive medications were recorded and correlation analysis among indexes was performed. Results The calf-RBI showed a normal distribution in both healthy subjects and HD patients. The calf-RBI was positively correlated with age, but not with gender or BMI. Forty-two patients with (35.9%) calf-RBI beyond target range were identified in 117 HD patients. The percentage of uncontrolled hypertensive individuals was significantly higher as compared to those with calf-RBI within or below target range (59.5% vs 33.3% and 16.7%, P＜0.01). The percentage of uncontrolled hypertensive individuals and the dose of antihypertensive medications was significant improved after decreasing the DBW in the patients with calf-RBI beyond target range (74.1% vs 33.3%, P＜0.01) and defined daily dose (2.00±2.28 vs 2.49±2.47, P＜0.05 ). Conclusions The age-stratified calf-RBI may be used as a useful index for estimation of volume status, and has a good association with clinical manifestations. Recognition and correction of chronic fluid overload based on age-stratified calf-RBI is helpful in hypertension control for hemodialysis patients.