Regulation of intracellular level of ATP and NADH in Escherichia coli to promote succinic acid production / 生物工程学报

Succinic acid is an important C4 platform chemical that is widely used in food, chemical, medicine sectors. The bottleneck of fermentative production of succinic acid by engineered Escherichia coli is the imbalance of intracellular cofactors, which often leads to accumulation of by-products, lower yield and low productivity. Stoichiometric analysis indicated that an efficient production of succinic acid by E. coli FMME-N-26 under micro-aeration conditions might be achieved when the TCA cycle provides enough ATP and NADH for the r-TCA pathway. In order to promote succinic acid production, a serial of metabolic engineering strategies include reducing ATP consumption, strengthening ATP synthesis, blocking NADH competitive pathway and constructing NADH complementary pathway were developed. As result, an engineered E. coli FW-17 capable of producing 139.52 g/L succinic acid and 1.40 g/L acetic acid in 5 L fermenter, which were 17.81% higher and 67.59% lower than that of the control strain, was developed. Further scale-up experiments were carried out in a 1 000 L fermenter, and the titer of succinic acid and acetic acid were 140.2 g/L and 1.38 g/L, respectively.

MicroRNA-138 Suppresses Adipogenic Differentiation in Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting Lipoprotein Lipase

PURPOSE: Adipogenic differentiation of adipose tissue-derived mesenchymal stem cells (AMSCs) is critical to many disease-related disorders, such as obesity and diabetes. Studies have demonstrated that miRNA-138 (miR-138) is closely involved in adipogenesis. However, the mechanisms affected by miR-138 remain unclear. This work aimed to investigate interactions between miR-138 and lipoprotein lipase (LPL), a key lipogenic enzyme, in AMSCs. MATERIALS AND METHODS: Human AMSCs (hAMSCs) isolated from human abdomen tissue were subjected to adipogenic differentiation medium. Quantitative real-time polymerase chain reaction and Western blot assay were applied to measure the expressions of miR-138, LPL, and the two adipogenic transcription factors cytidine-cytidine-adenosine-adenosine-thymidine enhancer binding protein alpha (C/EBPα) and peroxisome proliferator-activated receptor gamma (PPARγ). The relationship between miR-138 and LPL was predicted utilizing the miRTarBase database and validated by dual luciferase reporter assay. RESULTS: Showing increases in C/EBPα and PPARγ expression levels, hAMSCs were induced into adipogenic differentiation. During adipogenesis of hAMSCs, miR-138 expression was significantly downregulated. Overexpression of miR-138 by transfection inhibited hAMSCs adipogenic differentiation in vitro. Mechanically, LPL was a target of miR-138. LPL expression was upregulated during adipogenesis of hAMSCs, and this upregulation was reversed by miR-138 overexpression. Functionally, silencing of LPL by transfection exerted similar inhibition of the expressions of C/EBPα and PPARγ. Meanwhile, LPL ectopic expression was able to partly abolish the suppressive effect of miR-138 overexpression on adipogenic differentiation of hAMSCs. CONCLUSION: Upregulation of miR-138 inhibits adipogenic differentiation of hAMSCs by directly downregulating LPL.

The diagnostic value of PLASMIC score in the discrimination between thrombotic thrombocytopenic purpura and disseminated intravascular coagulation / 中华血液学杂志

Objective@#PLASMIC score was evaluated its value in differential diagnosis between the patients with thrombotic thrombocytopenic purpura (TTP) and those with disseminated intravascular coagulation (DIC) .@*Method@#Twenty-four patients with TTP and 41 cases with DIC were retrospectively analyzed in this study. The platelet count, average red blood cell volume, indirect bilirubin, creatinine and prothrombin time international normalised ratio were collected, and then PLASMIC scores were calculated.@*Results@#According to the risk classification of PLASMIC score, three (12.5%) TTP patients had moderate risk, and the rest 21 (87.5%) cases had high risk. In DIC patients, 92.7% cases were in low risk group, 4.9% at moderate risk, and only one case had high risk. Of these 65 patients, the sensitivity and the specificity to TTP of the high risk of the scoring system were 87.5% and 97.6%, respectively.@*Conclusion@#The patients with high risk of PLASMIC score correlated well with clinical TTP diagnosis. The scoring system showed to be an excellent diagnostic model to distinguish TTP patients from those with DIC.

Thrombelastography and conventional coagulation test for monitoring the perioperative coagulation state after joint arthroplasty / 中国组织工程研究

BACKGROUND:There is a general consensus that patients undergoing joint arthroplasty surgery wil be in hypercoagulable state and easily to induce deep vein thrombosis. Thromboelastography is a new kind of method to monitor blood coagulation state, but not widely used in orthopaedics. No final conclusion has yet been reached on whether we can guide the clinical prevention of deep vein thrombosis and medication through using thromboelastography to monitor perioperative coagulation state of patients treated with joint arthroplasty. OBJECTIVE:To investigate the correlation between thromboelastography and routine coagulation functional tests, and evaluate the clinical application value of thromboelastography in monitoring the perioperative coagulation state of patients treated with joint arthroplasty. METHODS:A total of 204 patients who treated with joint arthroplasty at First Affiliated Hospital of Soochow University from November 2014 to August 2015 were retrospectively analyzed. The thromboelastography, routine coagulation, platelet and other data before and after the replacement were respectively col ected. The correlative analysis was conducted between the thromboelastography result and the results of conventional coagulation test, that is, routine coagulation and platelet count. RESULTS AND CONCLUSION:In total knee arthroplasty group, activated partial thromboplastin time and reaction time showed good consistency (φ=0.713, Kappa value=0.647);Prothrombin time had moderate correlation and general consistency with reaction time (φ=0.392, Kappa value=0.362);Coagulation time and fibrinogen had moderate correlation and consistency (φ=0.392, Kappa value=0.488);Aggregates formation rate (αangle) and fibrinogen had moderate correlation and consistency;the remaining parameters had poor correlation and consistency. In total hip arthroplasty group, there was a weak correlation and consistency between the reaction time, activated partial thromboplastin time and prothrombin time;the other correlations were poor. However, there was a higher proportion of consistent clotting trend between some parameters of thrombelastography and routine coagulation. In total hip arthroplasty group, the consistent proportion of coagulation time and fibrinogen accounted for 67.6%;the consistent proportion of aggregates formation rate (αangle) and fibrinogen accounted for 78.3%. These results suggest that thromboelastography and routine coagulation tests have some correlations and consistency. Thromboelastography parameters have more consistent tendency on the data. Thrombelastography can serve as an auxiliary mean to monitor coagulation state of perioperative joint arthroplasty.

Effects of resina draconis on Toll-like receptor-4/nuclear factor-kappaB and dendritic cell phenotypes in colitis rats / 中国组织工程研究

BACKGROUND:Dendritic cel s can regulate the immunological reaction in the intestinal tract, this functional deficit may induce inflammatory bowel disease. Tol-like receptor-4/nuclear factor-κB pathway is highly involved in this reaction. OBJECTIVE:To establish experimental colitis model in rats, to observe effects of resina draconis on dendritic cel s and Tol-like receptor-4/nuclear factor-κB expression in rats with experimental colitis, and to explore its action mechanism. METHODS:A total of 44 male Sprague-Dawley rats were randomly assigned to four groups (n=11):blank control group, model group, resina draconis group, 5-aminosalicylic acid treatment group. With the exception of blank control group, 2,4,6-trinitrobenzenesulfonic acid-induced ulcerative colitis models were established in the model group, resina draconis group and 5-aminosalicylic acid treatment group. After the models were successful y established, the rats in the resina draconis and 5-aminosalicylic acid treatment groups were intragastrical y treated with resina draconis [(0.75 g(kg·d)] and 5-aminosalicylic acid [100 mg(kg·d)] respectively for 10 days. RESULTS AND CONCLUSION:Disease activity index, macroscopic colonic damage score and histopathological score were significantly decreased in the resina draconis group compared with the model group (P<0.05). Symptoms and tissue damages were obviously lessened in the 5-aminosalicylic acid treatment and resina draconis groups compared with the model group. Expression rates of CD80 and CD86, as wel as expression levels of Tol-like receptor-4 and nuclear factor-κB were significantly higher in the model group compared with the blank control group, resina draconis group and 5-aminosalicylic acid treatment group (P<0.05, P<0.01). Tol-like receptor-4 and nuclear factor-κB expression was significantly lower in the resina draconis group than that in the 5-aminosalicylic acid treatment group. Experimental findings indicate that, resina draconis can partial y relieve experimental colitis symptoms in rats and effectively inhibit the activation of dendritic cel s in the mesenteric lymph node. Resina draconis can relieve enteric inflammatory reaction by suppressing the expression of Tol-like receptor-4 and nuclear factor-κB in rats.

Comparison of biological characteristics of adipose-derived stem cells from different parts / 中国组织工程研究

BACKGROUND: Whether the differences exist between adipose-derived stem cells isolated from different parts of rats when cultured in vitro has been poorly understood. OBJECTIVE: To compare the growth characteristics and adipogenic ability of adipose-derived stem cells isolated from different parts of rats. METHODS: Freshly isolated adipose-derived stem cells were obtained from 5 mL inguinal groove and greater omentum adipose tissue of F344 rats using type Ⅰ col agenase digestion method. Then, adipose-derived stem cells were counted and cultured in vitro. Morphological and growth characteristics of adipose-derived stem cells derived from the two sites were observed. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was utilized to examine the doubling time of adipose-derived stem cells from different parts. The passage 2 adipose-derived stem cells were induced adipogenical y. Fourteen days after being induced, the differentiated cells were stained with oil red O and the positive cells were counted. The adipogenic differentiation ability of adipose-derived stem cells from the different parts was assessed. RESULTS AND CONCLUSION: (1) The number of adipose-derived stem cells from the greater omentum fat tissue in the same group was (281±10)×107/L, which was significantly higher than that from the inguinal groove fat tissue [(85±10)×107/L] (P < 0.01). Adipose-derived stem cells from the greater omentum and inguinal groove fat tissue achieved the exponential growth period on days 5 and 6, respectively, and achieved the platform period on days 9 and 10, respectively. The corresponding doubling time was 50 hours and 60 hours, respectively. After being passaged, adipose-derived stem cells grew in fibroblast-like shape actively. The adipogenic differentiation rate of adipose-derived stem cells from the greater omentum fat tissue was higher than that from the inguinal groove fat tissue [(38.90±2.86)% vs. (35.30±3.29)%, P < 0.01]. This shows that the number and the adipogenic differentiation ability of adipose-derived stem cells from different parts of the same F344 rat are different.

Correlation of serum uric acid levels with the progression of parkinson's disease / 中国综合临床

Objective To discuss the relationships between serum uric acid levels and clinical stages,cognitive dysfunction in parkinson' s disease (PD).Methods Sixty patients of PD were enrolled,and their history,detailed physical examination UPDRS Ⅲ scale and mini-mental state score (MMSE) scale were performed to determine the Hoehn-Yahr stage of patients.At the same time 52 cases of healthy person were collected from the Health Examination Centers whose age,sex,and education matched with PD group as control group.The levels of serum uric acid in the two groups were examined and compared.Results The serum uric acid levels in PD group were significantly lower than those in normal control group((262.4 ±85.2) μmol/L vs (357.6 ±73.3) μmol/L) (t =1.013,P =0.005) ;There was no level correlation between different Hoehn-Yahr stage period of PD group (r =0.143,P ＞ 0.05) ; There was no significant difference on serum uric acid levels between subgroup of PD patients (F =1.014,P =0.06) ;The M MSE score between PD group and normal control group had no significant difference (26.12 ± 2.87 vs 28.32 ± 2.17) (t =1.970,P =0.073).Conclusion Low level of the serum uric is likely to be a risk factor for PD,but cannot be used to assess the severity of disease and MMSE scale used to determine the cognitive impairment in PD is not sensitive enough.

Influence of mesenchymal stem cell transplantation on stereotypic behavior and dopamine of Tourette syndrome rats / 中华行为医学与脑科学杂志

ObjectiveTo investigate the effect of mesenchymal stem cell transplantation on stereotypic behavior and level of dopamine of Tourette syndrome(TS) rats.MethodsWistar rats were randomly divided into three groups namely sham group (microinfused with normal sera),TS group (microinfused with TS sera)group and TS plus MSC transplantation group.Auto-immune TS rat model were established and MSC suspension labeled with BrdU was injected into the striatum of TS rats.DA were measured by high performance liquid chromatography and electrochemical detection.ResultsTS rats with MSC grafts exhibited significantly decreased stereotypic behaviors (7d:117.3 ±8.4,14d:94.8 ±7.9,28 d:78.8 ±7.4)and level of DA in striatum(7 d:(898.2 ± 143.2) ng/g,14d:(812.4 ±92.2)ng/g,28 d:(757.9 ± 82.4)ng/g) at 7,14 and 28 days after transplantation compared to model group( ( 129.3 ± 9.8,116.5 ± 8.5,106.8 ± 6.4 ; ( 1209.3 ± 182.3 ) ng/g,( 1033.5 ± 114.7 ) ng/g,(994.8 ±112.6) ng/g,P＜0.01).DA in plasma of TS rats with MSC grafts also decreased at 7 and 14 days(7 d:(530.2 ±86.2)ng/g vs (690.3±72.7) ng/g;14d:(562.4 ±64.2) ng/g vs (689.4±83.9) ng/g,P＜0.01).ConclusionsTransplantation of MSC could reduce stereotypic behaviors in TS rats and regulate the metabolism of DA.

Clinical application of modified reduction plasty of malar and zygoma / 中华医学美学美容杂志

Objective To introduce a new method for correcting prominent malar complex deformity. Methods Through an intraoral incision, the highest area of zygomatic body marked preoperatively was grinded. Then an L shape incomplete osteotomy of the zygomatic body was performed with a reciprocating saw, and a complete osteotomy just 1 cm anterior to the articular tubercle of the zygomatic arch was made. Light pressure on the posterior part of the arch produced a greenstick fracture of the anterior osteotomy site, resulting in posterior-inward repositioning of the malar complex. Internal fixation was unnecessary. Results Operative procedures for reductive malar complex plasty were performed in 650 cases, which included 60 males and 590 females whose age ranged from 19 to 39 years.Incisions of all cases healed well. One case had maxillary sinusitis 2 weeks postoperatively, and recovered after 1 week by using antibiotics and drainage. There was 1 case with skin necrosis about 1 cm in diameter in the area of zygomatic body because of local liposuction, and the wound was healed by changing dressing. The forehead wrinkle of one side had disappeared in 1 case 1 week postoperatively, but had recovered 2 weeks later. Postoperative follow-up for 2-24 months showed satisfactory results.Conclusions This modified method has many advantages, such as simplicity, without internal fixation, short operation and recovery time, and little complications. The authors conclude that this technique is an effective and safe method of reduction malarplasty.

Preparation of Isoniazid-contained Drug Layer Alloy Stent and Its Dissolution in Vitro / 中国药房

OBJECTIVE:To prepare a nickel-titanium alloy stent coated by anti-tuberculotic isoniazid and study the dissolution of isoniazid in vitro.METHODS:The polyurethane which was used as film-former was mixed with isoniazid before being coated uniformly on the stent,then the stend was torrefied and dried so that the drug layer alloy stent was obtained.The content and accumulated dissolution in vitro of isoniazid from the stent was determined by ultraviolet spectrophotometry.RESULTS:The linear range of isoniazid was 5.0～30.0 ?g?mL-1(r=0.999 9)and its recovery rate was 99.67%(RSD=1.06%).A high accumluated dissolution rate of isoniazid was achieved,over 60% within the first 30 minutes,followed by a sustained dissolution in the following 8 h.CONCLUSION:The preparation method of the stent is simple and the quality is controllable.

Grafting of vascularized fibula and ilium to repair bone defects of limbs / 中华创伤骨科杂志

Objective To study the clinical results of grafting of vascularized fibular and ilium to repair bone defects of limbs. Methods From January 1992 to January 2004, 49 cases of bone defects of limbs were treated with vascularized fibular grafting (33 cases) and vascularized iliac grafting (16 cases). Their limb functions were graded according to Ennekings system. The changes of the vascularized fibular and iliac grafts were evaluated radiographically according to the International Symposium on Limb Salvage. Results All the patients were followed up for 1 to 12 years. In the vascularized fibular group, the recovery rate for the operated limbs was 82.7%, the average healing time of the bone was 16.8 weeks and the healing rate of the bone was 87.9%. In the other group, the recovery rate of the operated limbs was 88%, the average healing time of the bone was 15.2 weeks and the healing rate of the bone was 93.8%. Conclusion The vascularized fibular graft can be adopted to repair long bone defects of lower limbs and distal radius, while the vascularized iliac graft can be adopted to repair the bone defects of upper limbs.