Objective To purify rabbit anti-hRBBP10 polyclonal antibody.Methods The recombinant fusion protein PTC-hRBBP10 was expressed in the E.coli and was purified through amylose resin chromatography column and superose 12 gel fitration .The purified PTC-hRBBP10 was coupled to the NHS-activated sepharoseTM to prepare affinity chromatography column to purify rabbit anti-hRBBP10 polyclonal antibody. Results ① PTC-hRBBP10 was expressed and purified successfully with relative molecular mass(Mr) of 80?103 and its purity could reach about 95%.② Purified Rabbit anti-hRBBP10 polyclonal antibody could bind Specifically to PTC-hRBBP10.Conclusions With better specificity, The purified rabbit anti-hRBBP10 polyclonal antibody provide condition for studying the function of the protein, and helps to develop new techniques of tumor diagnosis and treatment.

Objective To explore the possibility of differentiation of canine bone marrow-derived mesenchymal stem cells(BMSCs) into smooth muscle cells(SMCs) and the potential of using these SMCs as cell sources for engineering of blood vessel construction.Methods Canine BMSCs were isolated by density gradient centrifugation and cultivated in DMEM supplemented with PDGF-BB and vitamin C(VIT-C).The phenotypic characteristics of BMSCs were identified by morphological observation,?-SMA and SMMHC in SMCs at passage 4-6 were analyzed by immunofluorescent staining,and the positive rates of SMCs at passage 5,6 were detected by flow cytometry.Results The BMSCs cultivated in the conditioned medium for SMCs showed SMC-like morphology:they displayed spindle-shape in morphology and were positive for ?-SMA but negative for SMMHC.42 d and 33 d were needed to obtain 107-108 seeding cells without or with PDGF-BB/VIT-C in culture medium respectively.With the subculture,the percentage of ?-SMA positive cells increased from 57.8% at passage 5 to 66.8% at passage 6,suggesting under aforementioned cultured conditions,more BMSCs turned into SMCs.Conclusion BMSCs can be differentiated into SMCs under appropriate culture conditions,suggesting the potentiality of using these SMCs as cell sources for tissue engineering of blood vessel construction.

Objective To establish rat models with pulmonary artery hypertension induced by monocrotaline(MCT) and to observe the pathological changes of lung tissue.Methods Seventy male Wistar rats were randomly devided into three group:control group(n=10),50 mg?kg-1 MCT group(n=30),60 mg?kg-1 MCT group(n=30).At two weeks and four weeks after injected intraperitoneally with MCT(injected intraperitoneally with equal normal saline in control group),the right ventricular systolic pressure(RVSP)and right ventricle weight/left ventricle+septum weight [RV/(LV+S)] ratio were measured.Hematoxylin-eosin staining and orcein technique were used to observe the pathological changes of lung tissue and pulmonary arterioles'medial thickness.Results Two weeks or four weeks after MCT administration,RVSP in 50 mg?kg-1 MCT group was higher than that in control group(respectively 36.6 mmHg?5.1 mmHg,39.1 mmHg?7.0 mmHg versus 26.1 mmHg?3.8 mmHg,both P

We isolated a new strain of endophytic Pseudomonas G5 from the stems of Chinese parsley (Coriandrum sativum L.), and it is tentatively identified as Pseudomonas aurantiaca according to analysis of the entire substrate utilization profiles using BIOLOG Microstation system (BIOLOG, Inc, Hayward CA). An array of evidence established that many Gram-negative bacteria employ Quorum sensing (QS) system to regulate gene expression in response to cell density using small diffusible signal molecules, N-acyl homoserine lactones (AHLs), and control diverse phenotypic traits in plant-associated bacteria. In this study, we showed that Pseudomonas sp. strain G5 can produce several types of AHLs at a detectable level using Thin Layer Chromatography (TLC) analysis combined with bioreporter Chromobacterium violaceum CV026 bioassay, and N-hexanoyl-homoserine lactone (HHL, C6-HSL) with Rf value 0.4 is the major signal molecule. Furthermore, we have identified its quorum sensing system composed of PhzI and PhzR by cloning and sequencing of phzI-phzR. PhzI is responsible for synthesis of AHLs signal molecules, and PhzR is a transcriptional regulator. Finally, we heterologously expressed the recombinant plasmid pMD-phzIR in Escherichia coli JM109 and verified it using C. violaceum CV026 bioassay. The phylogenetic analysis using MEGA4 revealed highly similarities exist among the phzIR homologs, suggesting it is evolutionary well conserved in the genus Pseudomonas.
4-Butyrolactone ; analogs & derivatives ; metabolism ; Acyl-Butyrolactones ; metabolism ; Amino Acid Sequence ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Phylogeny ; Pseudomonas ; classification ; genetics ; isolation & purification ; Trans-Activators ; biosynthesis ; genetics

Objective:To evaluate the effects of amelogenin(AML)on the migration,adhesion and proliferation of periodontal liga-ment stem cells(PDLSCs).Methods:PDLSCs were cultured with AML at 0.25,50 amd 100 μg/ml respectively.The migration, adhesion and proliferation of the cells were examined by wound healing migration assay,transwell migration assay,attachment assay, MTT assay and cell counting,respectively.Results:AML induced the migration of PDLSCs in a dose-dependent manner(P <0.05), increased the adhesion and proliferation of PDLSCs(P <0.05).Conclusion:AML may promote the migration,adhesion and prolifer-ation of PDLSCs.

Acute respiratory distress syndrome(ARDS) is a common critical syndrome with high morbidity and mortality.The treatments include respiratory support and drug therapy.In this paper, the latest clinical advances in the treatment of ARDS with glucocorticoids, statins, antioxidants, recombinant human activated protein C, sedatives and muscle relaxants, exogenous alveolar surfactants, mesenchymal stem cells, and vasodilators are reviewed.

Objective: To investigate the effect of dexmedetomidine on stress response and immune balance in patients with acute respiratory distress syndrome(ARDS) undergoing mechanical ventilation. Methods: From March 2016 to September 2018, 90 ARDS patients with mechanical ventilation in Chongqing Three Gorges Center Hospital were randomly divided into dexmedetomidine group(D group, 30 cases), propofol group(P group, 30 cases), midazolam group(M group, 30 cases) by random number table method.Another 30 healthy workers in our hospital were selected as normal control group(C group). In M, P and D groups, the sedative drugs were infused by micro-pump on the basis of analgesic treatment(fentanyl citrate). The heart rate(HR), mean arterial pressure(MAP), Murray score, acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score, plasma norepinephrine(NS), plasma cortisol(Cor), and blood T lymphocyte subsets(CD₄⁺, CD₈⁺, CD₄⁺/CD₈⁺) were observed in all patients at T0(the beginning of sedation), T1 (6h after sedation), T2 (24h after sedation), T3(48h after sedation). The levels of NS and Cor in plasma and blood T lymphocyte subsets(CD₄⁺, CD₈⁺, CD₄⁺/CD₈⁺) in C group were measured at 8∶00 a. m.the next day. Results: The plasma NS and Cor at T0 in M, P and D group were higher than those in C group (all P<0.05), while the levels of CD₄⁺ were lower than those in C group(all P<0.05). Compared with T0, the plasma NS and Cor decreased at T3 in M, P and D group (all P<0.05). The plasma Cor at T3 in P group was lower than that in M group[(475.77±49.69)μg/L vs.(502.33±60.92)μg/L](t=-26.57, P<0.05). The plasma NS and Cor at T3 in D group were lower than those in M group[(20.64±4.70)ng/L vs.(25.29±4.03)ng/L, (442.83±66.59)μg/L vs.(502.33±60.92)μg/L](t=-4.51, -59.50, all P<0.05), and were lower than those of P group[(20.64±4.70)ng/L vs.(23.60±4.49)ng/L, (442.83±66.59)μg/L vs.(475.77±49.69)μg/L](t=-2.96, -32.93, all P<0.05). Compared with T0, the level of CD₄⁺ decreased at T1, T2 and T3(except T3 in D group) in M, P and D group (all P<0.05), and the level of CD₈⁺ decreased at T1, T2 and T3 in M and P group (all P<0.05), but those in D group at T1, T2 and T3 were not significantly decreased(all P>0.05). The levels of CD₈⁺ at T2 and T3 in P and D groups were higher than that of M group (all P<0.05). The level of CD₄⁺/CD₈⁺ at T3 in D group were higher than that in M and P group[(1.46±0.30) vs.(1.26±0.33), (1.46±0.30) vs.(1.32±0.34)](t=0.21, 0.15, all P<0.05). Conclusion Sedation with dexmedetomidine or propofol can alleviate stress response and improve immune function in ARDS patients undergoing mechanical ventilation, and dexmedetomidine is better than propofol.

Objective To summarize and evaluate the clinical effect of extraco-rporeal membrane oxygenation(ECMO) support for critically ill patients after heart surgery,and to explore the factors that influenced the prognosis of ECMO.Methods The clinical data of 67 patients undergoing ECMO admitted to the Second Hospital of Jilin University from August 2013 to July 2017 were retrospectively analyzed.The patient's age ranged from 22 months to 78 years,including 3 infants aged 22,24,30 months and 64 adults ranged from 32 to 78 years,the mean age was(56.52 ± 10.99) years.Results The average ECMO support time was (235.79 ± 123.87) h,the mean mechanical ventilation time was (166.11 ± 148.31) h.49 patients weaned off successfully from ECMO,38 of them were discharged and 11 died within 28 days.18 patients gaved up treatment.42 patients suffered the complications.Conclusion VA-ECMO is a significant supportive method for preoperative intensive care in patients undergoing cardiovascular surgery.The keys to optimal results are grasping the indications of ECMO,earlier stopping mechanical ventilation,cluster curing(which involves reasonable anticoagulation,protection of organ function,prevention of hemorrhage,infection and limb distal ischemia).

Objective:To investigate the experience of moderate hypothermia(28℃) using in Stanford A aortic dissection.Methods:A retrospective analysis of 100 patients with Stanford A aortic dissection from January 2012 to March 2014, including 50 cases with deep(25℃) hypothermic circulatory surgery and 50 moderate(28℃) hypothermic circulatory surgery. The operation was performed with ascending aortic replacement, inclusiong aortic arch angioplasty, and stent implantation with descending aorta stent. The difference between the 2 groups during and after the operation was compared.Results:2 cases died in the hypothermia group, and 3 cases died in the deep hypothermia group. There were significant differences( P<0.05) between the middle and low temperature groups in the cooling time, the time of stopping circulation, the time of rewarming, the time of cardiopulmonary bypass, the time of operation, the time of operation, the time of conscious, the time of mechanical ventilation and the first day after the operation( P<0.05), but there was no significant difference between the creatinine and the bilirubin( P>0.05). Conclusion:Under the condition of sufficient cerebral perfusion and spinal cord protection, moderate hypothermia is safe, and it can reduce the operation time and postoperative complications. It has certain clinical significance.

Objective:To investigate the clinical characteristics of choledocholithiasis com-bined with periampullary diverticulum and influencing factor for difficult cannulation of endoscopic retrograde cholangiopancreatography (ERCP).Methods:The retrospective case-control study was conducted. The clinical data of 1 920 patients who underwent ERCP for choledocholithiasis in 15 medical centers, including the First Hospital of Lanzhou University, et al, from July 2015 to December 2017 were collected. There were 915 males and 1 005 females, aged (63±16)years. Of 1 920 patients, there were 228 cases with periampullary diverticulum and 1 692 cases without periampullary diverticulum. Observation indicators: (1) clinical characteristics of patients with choledocholithiasis; (2) intraoperative and postoperative situations of patients undergoing ERCP for choledocholithiasis; (3) influencing factor analysis for difficult cannulation in patients undergoing ERCP for choledocholithiasis. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range) or M( Q1, Q3), and com-parison between groups was conducted using the Wilcoxon rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. The Logistic regression model was used for univariate and multivariate analyses. Results:(1) Clinical characteristics of patients with choledocholithiasis. Age, body mass index, cases with complications as chronic obstructive pulmonary disease, diameter of common bile duct, cases with diameter of common bile duct as <8 mm, 8?12 mm, >12 mm, diameter of stone, cases with number of stones as single and multiple were (69±12)years, (23.3±3.0)kg/m 2, 16, (14±4)mm, 11, 95, 122, (12±4)mm, 89, 139 in patients with choledocholithiasis combined with periampullary diverticulum, versus (62±16)years, (23.8±2.8)kg/m 2, 67, (12±4)mm, 159, 892, 641, (10±4)mm, 817, 875 in patients with choledocholithiasis not combined with periampullary diver-ticulum, showing significant differences in the above indicators between the two groups ( t=?7.55, 2.45, χ2=4.54, t=?4.92, Z=4.66, t=?7.31, χ2=6.90, P<0.05). (2) Intraoperative and postoperative situations of patients undergoing ERCP for choledocholithiasis. The balloon expansion diameter, cases with intraoperative bleeding, cases with hemorrhage management of submucosal injection, hemostatic clip, spray hemostasis, electrocoagulation hemostasis and other treatment, cases with endoscopic plastic stent placement, cases with endoscopic nasal bile duct drainage, cases with mechanical lithotripsy, cases with stone complete clearing, cases with difficult cannulation, cases with delayed intubation, cases undergoing >5 times of cannulation attempts, cannulation time, X-ray exposure time, operation time were 10.0(range, 8.5?12.0)mm, 56, 6, 5, 43, 1, 1, 52, 177, 67, 201, 74, 38, 74, (7.4±3.1)minutes, (6±3)minutes, (46±19)minutes in patients with choledocholithiasis combined with periampullary diverticulum, versus 9.0(range, 8.0?11.0)mm, 243, 35, 14, 109, 73, 12, 230, 1 457, 167, 1 565, 395, 171, 395, (6.6±2.9)minutes, (6±5)minutes, (41±17)minutes in patients with choledocholithiasis not combined with periampullary diverticulum, showing significant differences in the above indicators between the two groups ( Z=6.31, χ2=15.90, 26.02, 13.61, 11.40, 71.51, 5.12, 9.04, 8.92, 9.04, t=?3.89, 2.67, ?3.61, P<0.05). (3) Influencing factor analysis for difficult cannulation in patients undergoing ERCP for choledocholithiasis. Results of multivariate analysis showed total bilirubin >30 umol/L, number of stones >1, combined with periampullary diverticulum were indepen-dent risk factors for difficult cannulation in patients with periampullary diverticulum who underwent ERCP for choledocholithiasis ( odds ratio=1.31, 1.48, 1.44, 95% confidence interval as 1.06?1.61, 1.20?1.84, 1.06?1.95, P<0.05). Results of further analysis showed that, of 1 920 patients undergoing ERCP for choledocholithiasis, the incidence of postoperative pancreatitis was 17.271%(81/469) and 8.132%(118/1 451) in the 469 cases with difficult cannulation and 1 451 cases without difficult cannula-tion, respectively, showing a significant difference between them ( χ2=31.86, P<0.05). In the 1 692 patients with choledocholithiasis not combined with periampullary diverticulum, the incidence of postopera-tive pancreatitis was 17.722%(70/395) and 8.250%(107/1 297) in 395 cases with difficult cannula-tion and 1 297 cases without difficult cannulation, respectively, showing a significant difference between them ( χ2=29.00, P<0.05). In the 228 patients with choledocholithiasis combined with peri-ampullary diverticulum, the incidence of postoperative pancreatitis was 14.865%(11/74) and 7.143%(11/154) in 74 cases with difficult cannulation and 154 cases without difficult cannulation, respectively, showing no significant difference between them ( χ2=3.42, P>0.05). Conclusions:Compared with patients with choledocholithiasis not combined with periampullary divertioulum, periampullary divertioulum often occurs in choledocholithiasis patients of elderly and low body mass index. The proportion of chronic obstructive pulmonary disease is high in choledocholithiasis patients with periampullary diverticulum, and the diameter of stone is large, the number of stone is more in these patients. Combined with periampullary diverticulum will increase the difficult of cannulation and the ratio of patient with mechanical lithotripsy, and reduce the ratio of patient with stone complete clearing without increasing postoperative complications of choledocholithiasis patients undergoing ERCP. Total bilirubin >30 μmol/L, number of stones >1, combined with periampullary diverticulum are independent risk factors for difficult cannulation in patients of periampullary diverticulum who underwent ERCP for choledocholithiasis.