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Objective:To investigate the effects of alum ice nanoemulsion on VEGF and TGF-β1 in hypertrophic scar based on Notch signaling pathway.Methods:Totally 144 SD rats were divided into blank control group, model group, triamcinolone acetonide group and alum ice nanoemulsion low-, medium- and high-dose groups according to random number table method, with 24 rats in each group. Except for the blank control group, the rats in other groups were prepared with deep Ⅱ ° burn models. 24 hours after the successful modeling, the model group was given the same amount of normal saline, the rats in alum ice nanoemulsion low-, medium- and high-dose groups were given 8.15, 6.30 and 32.60 mg/ml alum ice nanoemulsion respectively, and the triamcinolone acetonide group was given triamcinolone acetonide twice a day, 0.2 ml each time, for 35 consecutive days. At 14, 21, 28 and 35 d, the collagen fiber surface density was calculated by VG staining. The protein expressions of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), Notch1 and Jagged1 were detected by Western Blot. The expressions of Notch1 mRNA and Jagged1 mRNA were detected by RT-PCR.Results:Compared with model group, triamcinolone acetonide and different doses of alum ice nanoemulsion groups could decrease collagen fiber surface density, protein expressions of VEGF, TGF-β1, Notch1, Jagged1 and mRNA expressions of Notch1, Jagged1 in different degrees ( P<0.05). Compared with the triamcinolone acetonide group, the collagen fiber surface density, protein expressions of VEGF, TGF-β1, Notch1 and Jagged1 and mRNA expressions of Notch1, Jagged1 in the alum ice nanoemulsion medium-dosage group decreased ( P<0.05). Conclusion:Alum ice nanoemulsion can inhibit hypertrophic scar formation, and its mechanism is related to down-regulating Notch signal pathway related molecules Notch1, Jagged1 protein and mRNA levels, and then down-regulating VEGF and TGF-β1 protein expressions.
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The modern bio-fermentation industry requires design and creation of efficient microbial cell factories for directed conversion of raw materials to target products. The main criteria for assessing the performance of microbial cell factories are their product synthesis capacity and stability. Due to the deficiencies of plasmids in gene expression such as instability and being easy to lose, integration of genes into chromosome is often a better choice for stable expression in microbial hosts. To this end, chromosomal gene integration technology has received much attention and has developed rapidly. In this review, we summarize the recent research progresses of chromosomal integration of large DNA fragments in microorganisms, illustrate the principles and features of various technologies, highlight the opportunity brought by the CRISPR-associated transposon systems, and prospect future research direction of this technology.
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Chromosomes , Plasmids , DNA , Cloning, Molecular , FermentationABSTRACT
OBJECTIVES@#Hypertrophic scar (HS) is the most common pathological scar in clinical practice. During its formation, angiogenesis-related factors show dynamic expression. Modern studies have found that Notch signaling pathway has an extremely important role in maintaining the construction and remodeling of vascular endothelial cells and vascular network. The correlation between Notch signaling pathway and angiogenesis in hypertrophic scar has been rarely reported. This study aims to investigate correlation between Notch signaling pathway and the expression of angiogenic factors in a proliferative scar model.@*METHODS@#A total of 81 Sprague Dawley rats (SPF grade) were randomly assigned into a blank control group, a model group, and a blocker group. In the blocker group, a 2 cm diameter circular scald head was placed on the back of the rats for 10 s at 75 ℃ by using a constant temperature and pressure electrothermal scalding apparatus to form a rat deep II° burn model, and a hyperplastic scar model rat was obtained after natural healing of the wound skin (21 to 23 day epithelialization). A syringe was used to inject a needle from the normal skin around the scar at the 1st, 3rd, 5th, 7th, and 14th days after modeling. The γ-secretase inhibitor was injected locally at 2 mg/kg in a dilution of 0.1 mL at the base of the scar. The rats in the model group was injected with the same amount of saline after modeling; the rats in the blank control group was injected with the same amount of saline. Nine rats in each group was randomly killed by air embolization at the 21st, 28th, and 35th days, respectively. The protein expressions of collagen type I (COL-I) and collagen type III (COL-III) were detected by immunohistochemistry. The protein expressions of vascular endothelial growth factor (VEGF), angiopoietin 1 (Ang1), transforming growth factor-β1 (TGF-β1), and matrix metalloproteinase-2 (MMP-2) were detected by Western blotting.@*RESULTS@#Immunohistochemical results showed that, at the 21st,28th, and 35th days, the protein expressions of COL-I and COL-III in the model group were up-regulated compared with the blank control group (all @*CONCLUSIONS@#In the Sprague Dawley rat proliferative scar model, inhibition of Notch signaling pathway could attenuate the expressions of COL-I and COL-III, reduce traumatic scar proliferation, down-regulate the expressions of VEGF, Ang1, TGF-β1, and MMP-2, and inhibit angiogenesis. The expressions of angiogenesis-related factors appeare to be up-regulated during the formation of proliferative scar. When the Notch signaling pathway is inhibited, the up-regulated angiogenic factors show a decreasing trend and the proliferative scar is alleviated, which suggests that Notch signaling pathway may affect the formation of hyperplastic scar by regulating the expression of angiogenic factors.
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Animals , Rats , Cicatrix, Hypertrophic/pathology , Endothelial Cells , Matrix Metalloproteinase 2 , Rats, Sprague-Dawley , Signal Transduction , Transforming Growth Factor beta1 , Vascular Endothelial Growth Factor AABSTRACT
Objective:To study the feasibility of establishing gastrointestinal anastomosis by magnetic compression technique in rabbits.Methods:Ten healthy New Zealand rabbits were selected as models for gastrointestinal anastomosis by magnetic compression technique. Daughter and parent magnets suitable for gastrointestinal anastomosis in rabbits were designed and manufactured. A daughter magnet was inserted into the stomach through the purse fistula in the lack of blood vessel area of gastric body, and was pushed into the duodenum along the intestinal tract. And then a parent magnet was inserted through the stomach fistula. The daughter and parent magnets were automatically attracted and pressed the gastric and intestinal walls after they were adjusted in the proper position. The stomach fistula was closed with purse string suture. After ischemia, necrosis, detachment of the tissues between magnets, gastrointestinal anastomosis was established, and the magnets and necrotic tissues were expelled together from the body through the digestive tract. Survival of experimental animals was observed. Anastomotic specimens were obtained one month after operation. The blasting pressure of anastomotic stoma was measured, and the healing of anastomotic stoma was observed with naked eyes.Results:According to the pre-designed operative route, 10 New Zealand rabbits all successfully completed the operation and survived one month after surgery. No complications occurred during perioperative period. The operation time was 35.80±4.71 min (range 28.00-43.00 min), and the magnet discharge time was 11.40±1.56 days (range 9.00-14.00 days). Anastomotic specimens were obtained one month after the operation. Gross observation showed that the anastomotic stoma of gastrointestinal bypass anastomosis healed well, and the surrounding tissues adhered slightly. The anastomotic bursting pressure was 103.00±7.95 mmHg (range 94.00-113.00 mmHg) (1 mmHg=0.133 kPa).Conclusion:The establishment of gastrointestinal anastomosis by magnetic compression technique in rabbits is simple and effective.
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Objective:To design magnets to locate colorectal neoplasms based on magnetic tracer technology, and to test its feasibility and safety by animal experiments.Methods:The magnets used for endoscopic localization of colorectal tumors consist of a tracer magnet and a pursuit magnet, both of which are ring-shaped Nd-Fe-B magnets. Eight healthy Beagle dogs were used as animal models. Tumor locations were assumed in the different parts of the colon and rectum under colonoscopy. The tracer magnet was sent to the hypothetical tumors by endoscopic soft tissue clamp and fixed near the tumors. After 24 hours, laparoscopic surgery was performed under general anesthesia. The pursuit magnet was inserted near the resected colon or rectum through the main operating hole. The tracer magnet was absorbed to the pursuit magnet to identify the location of tumors.Results:The tracer magnet and pursuit magnet were successfully designed and processed. The suction force between the tracer magnet and the pursuit magnet at zero distance was 16 N. All the 8 Beagle dogs successfully received indwelling of magnets under colonoscopy, and no magnets fell off after 24 hours. After the placement of pursuit magnet under laparoscopy, the two magnets attracted each other rapidly and accurately, and successfully completed localization of tumor site without any damage during the operation.Conclusion:Colonoscopy combined with laparoscopy for colorectal neoplasms localization based on magnetic tracer technique is simple, accurate, safe and feasible.
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Objective To verify the feasibility and safety of stomach tumor marker localization based on magnetic tracer technique in dogs.Methods Six male Beagle dogs were examined by gastroscopy.Then tracer magnets were sent to the "tumor" locations assumed in advance and fixed near the "tumors" by endoscopic soft tissue clamp.Laparoscopic gastric tumor localization was performed under general anesthesia 24 hours later.The tracer magnet was placed near the tumor on the surface of the stomach through the operating hole after the conventional establishment of laparoscope puncture parallel mirror to explore the tracer magnet.After the two magnets were attracted,the location of the tracer magnet seen under the laparoscope was the location of the gastric tumor,so as to complete the labeling and positioning of the lesion.Results All the 6 Beagle dogs were successfully implanted with tracer magnets under gastroscopy.Twenty-four hours after the gastroscopy,the pursuit magnet was successfully implanted during laparoscopic surgery.The two magnets automatically attracted each other and formed a sandwich structure of "tracer magnet-gastric wall-pursuit magnet ",which completed the location and identification of gastric tumor under the laparoscopy.Conclusion Gastroscopy combined with laparoscopy based on magnetic tracer technique is simple,accurate,safe and feasible.
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Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS), and explore the efficacy of endoscopic treatment in patients with esophageal submucosal tumor. Methods Sixty-eight patients with esophageal submucosal tumor were selected, and the tumor was derived from the muscularis mucosa and submucosa according to the common endoscope and endoscopic ultrasonography detection. Endoscopic mucosal resection (EMR) was applied to remove submucosal tumor with diameter less than 1.0 cm, endoscopic piecemeal mucosal resection (EPMR) or endoscopic submucosal dissection (ESD) was applied to remove submucosal tumor with diameter 1.1 - 1.5 cm, and ESD was applied to remove submucosal tumor bigger than 1.5 cm. Samples were examined by pathology after treatment. Results Tumors in all the patients were completely removed, and the tumor diameter was 0.6-2.3 cm. Forty-one cases were treated with EMR, 9 cases were treated with EPMR and 18 cases were treated with ESD. Four patients had intra-operative bleeding that was stopped by electrocoagulation hemostasis. No perforation occurred in all cases. Postoperative pathology revealed 43 cases had leiomyoma, 23 cases had interstitialoma, and 2 cases had lipoma. Patients were reviewed by gastroscope 3 months after operation. The white scars formed in all patients, and there was no residue or recurrence. Conclusions Different origin layers and property of esophageal submucosal tumor can be diagnosed accurately by EUS, and endoscopic therapy (EMR, EPMR and ESD) is an effective treatment for submucosal tumor from muscularis mucosa and submucosa. Endoscopic therapy is safe and effective. It provides sufficient pathological information.
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Objective:To prepare a bone substitute using microsphere scaffold containing adiponectin(APN)and to investigate the release behavior of the scaffold in vitro.Methods:Chitosan microsphere was developed by an emulsion-ionic cross-linking method. Poly (L-lactic-co-glycolic)acid (PLGA)and β-tricalcium phosphate (β-TCP)were used to prepare microsphere scaffold containing APN.The morphology,particle size,drug loading,incorporation efficiency and release behavior of the microsphere were examined. Results:The APN containing microsphere showed good spherical geometry,suitable size and microporosity under scanning electron microscope.The average diameter of the milipore was 20 -200 μm;the drug loading and incorporation efficiency were 1 .3% and 70.3% respectively.The controled-release process continued for 91 days.The extract solution from the APN microsphere-scaffold promoted MC3T3 cell proliferation without cytotoxicity.Conclusion:The APN microsphere-scaffold has sustained release function and may promote osteoblast proliferation.
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Objective To investigate the clinical effect of gastric bypass operation on treating type 2diabetes,and analyze the management status.Methods Fifty-seven patients wiith type 2 diabetes treated by gastric bypass operation were divided by different body mass index(BMI) into 4 groups:group A( BMI ＜ 20kg/m2) with 8 cases,group B (20 kg/m2≤BMI ＜24 kg/m2) with 21 cases,group C (24 kg/m2≤BMI ＜27kg/m2) with 17 cases and group D (BMI ≥27 kg/m2) with 11 cases.The control of blood glucose and glycosylated hemoglobin,changes of diets and body weight,diabetes complications,adverse reactions after operation and postoperative management were followed up or reviewed.The clinical data were analyzed comprehensively.Results The cure rate of group C was the highest [ 70.6％( 12/17 ) ],while the cure rate of group A was the lowest (0).The overall cure rate was 43.9％(25/57) and the overall improvement rate was 28.1％(16/57).The overall efficient rate was 71.9％(41/57) and the overall inefficient rate was 28.1％(16/57).Apart from a few patients had malnutrition,amenorrhea,bad stomach,diarrhea and poor diet after leaving the hospital,other patients had no surgery complications and no patients died.There was no clinician to follow up and did education to these patients after operation.Conchusions Gastric bypass operation is safe and effective for obese and non-obese patients with type 2 diabetes.It is a new method to weat type 2diabetes at present,but the indications should be grasped and operation enlargement must he prevented.The management of teams of physicians,surgeons,dietitians,nurses should be established afler operation.
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Objective:To investigate the distribution characteristics of syndrome types of traditional Chinese medicine (TCM) in essential hypertension and to explore the distribution rule of TCM syndromes. Methods: A multicenter, large-sample survey method of clinical epidemiology was applied to choose the patients with essential hypertension from North, Middle, and South China. A questionnaire was designed and filled in, then 477 untreated patients with first-diagnosed essential hypertension were selected and the information was recorded into FileMaker database. A cluster analysis method was utilized to study the TCM syndrome distribution rule of essential hypertension. Results: Two-step cluster analysis was done from 3 to 7 clusters. Seven clusters were appropriate, which included deficiency of heart and kidney qi, hyperactivity of liver-yang, deficiency of yin and yang, stagnation of phlegm-dampness, phlegm-heat (subtype of stagnation of phlegm-dampness), blood stasis obstructing collaterals, and other syndromes. The symptoms presenting high percentage in each cluster were more significant in TCM theory. The syndromes of hyperactivity of liver-yang (24.1%) and stagnation of phlegm-dampness (27.1%) presented the high percentages, and deficiency of heart and kidney qi (10.1%), deficiency of yin and yang (8.4%), and blood stasis obstructing collaterals (9.0%) presented the low percentages. Conclusion: As compared with the current syndrome differentiation criteria, two-step cluster analysis results not only include the syndromes of deficiency of yin and yang, hyperactivity of liver-yang, stagnation of phlegm-dampness, but also cover qi deficiency and blood stasis.
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Objective To analyze and investigate endoscopic and pathological characteristics in the diagnosis of early esophageal carcinoma. Methods The clinical data of 62 patients with early esophageal carcinoma who had been admitted to Southwest Hospital from January 2003 to December 2007 were retrospectively analyzed. Results Of all patients, 8 had upper esophageal carcinoma, 42 had middle esophageal carcinoma and 12 had lower esophageal carcinoma. The tumor growth patterns included surface diffusion growth (n = 30), bidirectional growth (n = 11), intracavitary growth (n = 9), intra-esophageal wall growth (n = 7) and mixed growth (n = 5). The diameter of lesions ranged from 1.0 cm to 2.9 cm in majority. Surgical resection was done in 38 patients, endoscopic mucosal resection in 23 patients and endoscopic submucosal dissection in 1 patient. Postoperative pathological examination showed that 13 patients had in situ carcinoma, 22 had intramucosal carcinoma and 27 had submucosal carcinoma. The lymph node metastatic rate of intramucosal carcinoma and submucosal carcinoma were 5% (1/22) and 15% (4/27), respectively. The numbers of squamous cell carcinoma, adenocarcinoma, small cell carcinoma, sarcoma carcinoma and spindle carcinoma were 57, 2, 1, 1, 1, respectively. ConelusionsEarly esophageal carcinoma tends to appear in the middle part of esophagus, and with surface diffusion growth type. Most of the early esophageal carcinoma are squamous cell carcinoma. Endoscopy combined with lugol's iodine and methyleneblue staining is effective in detecting early esophageal carcinoma.
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Blood stasis is one of the main pathogenesis of geriatrics.The main traits of geriatrics blood stasis are: blood stasis mainly induced by def iciency,def iciency and stasis is concomitant,stasis builds up gradually,symptoms of blood stasis aren’t mandatory in clinic practice,the range and severity of blood stasis is so wide and severe that it’s hard to be dissipated.Activating circulation to dissipate stasis is followed through the whole therapeutic process,and often used with replenishing qi,invigorating the kidney,and nourishing blood in order to dissipate stasis without hurting vital qi as well as reinforcing vital energy to dissipate stasis.
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Objective To prepare in-house calibrator of apolipoprotein A1 and apolipoprotein B with the values are traceable to the WHO-International reference materials SP1-01 and SP3-07.Methods We participated in the Northwest lipid research laboratory (NWLRL) protocol for target value transfer of WHO-IFCC apo A1 and apoB reference materials, and evaluated of comparability of the measurement.The analyses were performed on a Olympus AU 400, apo A1 and apoB were determined by immunoturbidimetric endpoint method.Results Through 3-step study, the precision and accuracy of apo A1 and apoB determination of our company met the NWLRL criteria.Correlation coefficients between obtained and assigned values of NWLRL on 40 individual serum samples were 0.983 for apo A1, 0.987 for apoB, the average absolute bias were 2.7 % for apo A1, 3.0 % for apoB, respectively.Conclusion NWLRL issued the certificate to Zhongsheng Beikong Bio-Technology and Science Inc.for the in-house apo A1 and apoB calibrator values are traceable to the WHO- International reference materials.