ObjectiveTo investigate the effect of cytoskeleton modification on the adipogenic differentiation of rat Achilles-derived tendon stem cells (TSCs) in vitro. MethodsTSCs were isolated from the tendon tissue of male Sprague Dawley rats (aged 3 weeks) by enzymatic digestion method and cultured for 3 passages. After the 3rd passage cells were cultured with DMEM medium containing 15% fetal bovine serum and cytochalasin D (CYD) at the concentrations of 0, 50, 100, 500, and 1 000 ng/mL, the cell survival condition and morphology changes were observed by inverted phase contrast microscope, the cytoskeleton was observed through fibrous actin (F-actin) staining, and the ratio of F-actin/soluble globular actin (G-actin) was detected and calculated through Western blot. According to the above results, the effective concentration of CYD was selected and used for next experiments. After TSCs were cultured for 3 and 7 days respectively with adipogenic induction media (induction group), adipogenic induction media containing CYD (CYD+induction group), ordinary medium (ordinary group), and ordinary medium containing CYD (CYD+ordinary group), the real-time quantitative PCR (qRT-PCR) and Western blot were carried out to measure the mRNA and protein expressions of adipogenic differentiation-related markers, including peroxisome proliferator-activated receptor γ (PPARγ), 1ipoprotein lipase (LPL), and fatty acid binding protein (aP2). ResultsThe final CYD concentration of 100 ng/mL can inhibit effectively G-actin polymerization into F-actin, but could not affect TSCs survival, which was used for next experiments. qRT-PCR and Western blot suggested that the mRNA expressions of PPARγ, LPL, and aP2 and the protein expressions of PPARγ and aP2 were increased significantly in the CYD+induction group at 3 and 7 days when compared with the induction group (P<0.05). In the CYD+ordinary group, there still was a significant increase in the mRNA expressions of PPARγ, LPL, and aP2 when compared with the ordinary group (P<0.05). ConclusionInhibition of F-actin polymerization can increase adipogenic differentiation of rat Achilles-derived TSCs in vitro, and cytoskeleton modification is a pre-requisite for TSCs differentiation into adipocytes, which might have important implications for the mechanism research of tendinopathy.
Objective To analyze the rationality of emergency medication in the West China Hospital after Lushan earthquake based on the actual drug use of earthquake victims. Methods We applied DDDs and DUI as evaluation index, input data using Excel software, and analyzed if the emergency medication was required for the injury/illness and the rationality of emergency medication. Results Earthquake victims mainly had trauma and wound infection and they were given antibiotics as main treatment one week after the earthquake. Drugs for the respiratory system and digestive system were mainly used in patients who mainly manifested as non-traumatic diseases and internal diseases four weeks after the earthquake. Among 49 kinds of drugs which could be calculated for the value of DUI, injection accounted for a larger proportion than non-injection (59.18% vs. 40.82%). The results showed that, the medication (DUI=1) only accounted for 12.24%; the medication (DUIlt;1, Mean=0.65) accounted for 38.78%, which implied insufficient drug use; the medication (DUIgt;1, Mean=1.77) accounted for 46.94%, which implied drug overuse. Conclusion Medication for every system is basically timely and rational when treating symptoms and causes in the West China Hospital within one month after Lushan earthquake. However, the rationality of emergency medications using DUI=1 under normal conditions still needs to be further verified.
Objective To analyze drug use for 23 sick and wounded hospitalized in the departments of gynaecology, obstetrics and paediatrics in the West China Second University Hospital, during one month after Lushan earthquake, so as to provide references for the drug use for the sick and wounded women and children. Methods Medication information was retrospectively investigated in the departments of gynaecology, obstetrics and paediatrics in the West China Second University Hospital during one month after Lushan earthquake. We extracted data including hospitalization duration, medication prescriptions, types and costs of essential medicines, types and costs of antibiotic medicines using the HIS system. Discharge diagnosis was standardized and classified according to ICD-10. Indicators of rational drug use included defined daily dose (DDD), defined daily dose statistic (DDDs), drug utilization index (DUI), children defined daily dose (cDDD), children drug utilization index (cDUI), proportion of essential medicines, proportion of antibiotic medicines, mismatch of medicine and diagnosis, repeated drug use, improper combination of drug, incorrect usage and dosage, and frequently drug change. Medication information was evaluated and discussed by the prescription working group. Discrepency was resolved by consulting with the panel. Results A total of 23 sick and wounded women and children were assessed during one month after Lushan Earthquake. There were 13 sick and wounded children in the department of pediatrics (mainly manifested as bronchial pneumonia) and 10 sick and wounded women in the department of gynaecology and obstetrics (mainly involved in cesarean delivery). Proportions of essential medicines were 13/13 in the department of pediatrics and 9/10 in the department of gynaecology and obstetrics. Proportions of antibiotic medicines were 10/13 in the department of pediatrics and 8/10 in the department of gynaecology and obstetrics. Irrational drug use was mainly caused by improper use of antibiotics. Conclusion The drug use for 23 hospitalized sick and wounded is rational in the departments of gynaecology, obstetrics and paediatrics in the West China Second University Hospital during one month after Lushan earthquake. Evaluation methods of rational drug use in high-risk population are urgently needed. Using cDDD and cDUI is an active exploration for the evaluation of pediatric drug use, which needs more practice to test and improve.
Objective To investigate the spectrum of diseases and the current situation of antibiotic use in rural hospitals and community health service centers in Chengdu, so as to provide evidence for selecting essential medicines and promoting rational use of antibiotics. Method We selected 7 township/community health institutions, from which we collected inpatient and outpatient information. Information about antibiotic use was also collected, including categories, cost, and dosage. A standard questionnaire was used to investigate physicians’ prescription behavior for principal diseases. Result Urban and rural areas had different spectrums of diseases. The major diseases in urban areas included diabetes mellitus, hypertension, chronic obstructive pulmonary disease, and respiratory tract infection; while those in rural areas were infectious diseases of the respiratory system, digestive system, and urinary system. The physicians’ prescription behavior was mainly based on their personal experience. Antibiotics accounted for 30-50% of the total medicine cost. The top four types of antibiotics with the highest cost were cephalosporins, penicillin, quinolones, and macrolides. Conclusion Based on the different spectrums of diseases, essential drug lists and standard treatment guidelines appropriate for rural health care should be developed to improve the rational use of drugs. Factors such as the average cost of daily dose and the course of treatment should be taken into consideration to reduce the overall cost of medicine. An antimicrobial resistance monitoring system and special training courses on rational use of antibiotics should be utilized in the rural health institutions.
Objective To explore the effect and mechanism of sleeve gastrectomy （SG） for type 2 diabetes mellitus （T2DM） in Goto-Kakizaki （GK） rats. Methods Thirteen male GK rats at 12 weeks of age were randomly divided into SG group （n=7） and sham operation group （SO group， n=6）， receiving SG surgery and sham operation respectively.Body weight， food intake in 24hours， fasting plasma glucose， plasma glucagon-like peptide-1 （GLP-1）， and plasma Ghrelin of rats in 2 groups were measured or tested before operation， 1， 4， 10， and 26 weeks after operation. In 10 weeks after operation， fecal energy content of rats in 2 groups was tested， in addition， oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were performed to investigate the glucose tolerance and insulin sensitivity. Results ①Body weight：there were no significant difference on body weight between the 2 groups （P＞0.05）. Compared with time point of before operation， the body weight of both 2 groups decreased in 1 week after operation （P＜0.01）， but increased in 10 weeks and 26 weeks （P＜0.01）. ②Food intake in 24 hours：compared with SO group， the food intake of SG group were lower in 4 weeks and 10 weeks after operation （P＜0.05）. Compared with time point of before operation， the food intake of SG group were lower in 1， 4， and 10 weeks after operation （P＜0.05）， but lower only in 1 week in SO group （P＜0.05）. ③Value of fasting glucose：compared with SO group， the value of fasting glucose in SG group were lower after operation （P＜0.01）. Compared with time point of before operation， the value of fasting glucose of SG group were lower after operation （P＜0.01）， but decreased in 1 week only in SO group （P＜0.01）. ④Level of serum GLP-1：compared with SO group， the levels of serum GLP-1 in SG group were higher in 4， 10， and 26 weeks after operation （P＜0.05）. Compared with time point of before operation， the levels of serum GLP-1 in SG group were higher in 4， 10， and 26 weeks after operation （P＜0.05）， but levels of serum GLP-1 in SO group didn’t change significantly （P＞0.05）. ⑤Level of serum Ghrelin：compared with SO group， the levels of serum Ghrelin in SG group were lower at alltime points after operation （P＜0.01）. Compared with time point of before operation， the levels of serum Ghrelin in SGgroup were lower at all time points after operation （P＜0.001）， but levels of serum Ghrelin in SO group didn’t change significantly （P＞0.05）. ⑥Areas under curves （AUC）：the AUC of OGTT and ITT test in SG group were both lower than those of SO group （P＜0.01）. Conclusion SG surgery can induce the level of fasting plasma glucose， and canimprove glucose tolerance and insulin sensitivity with significant changes of levels of plasma GLP-1 and Ghrelin， sugg-esting that SG surgery may be a potential strategy to treat patient with T2DM but without obesity or insulin resistance.
Objective To establish different kinds of reduced size liver transplantation model of rats， and to explorethe optimal marginal size of liver graft in orthotopic liver transplantation， in purpose of providing a kind of animal modelfor the study about mechanism and prevention measures of small-for-size syndrome. Methods One hundred and ninety-two rats were randomly divided into whole liver graft transplantation group （underwent whole liver graft transplantation），half liver graft transplantation group （the median lobe and right lobe of the liver were selected to be the graft）， small size liver graft transplantation group （the median lobe of the liver was selected to be the graft）， and extra-small size liver graft transplantation group （the median lobe and left lobe of the liver were reduced， and remained lobes were selected to be the graft）， each group enrolled 48 rats. After liver graft transplantation， 24 rats of each group were selected to observe the survival situation， 12 rats of each group were selected to measure portal venous pressure at time point of before operation，and 5， 15， 30， 45， and 60 minutes after transplantation. The other 12 rats of each group were test the level of alanine aminotransferase （ALT）. Results Seven-day survival rate of the whole liver graft transplantation group， half liver graft transplantation group， small size liver graft transplantation group， and extra-small size liver graft transplantation group was 100% （24/24）， 87.5% （21/24）， 37.5% （9/24）， and 0 respectively. Portal venous pressure of whole liver graft trans-plantation group was stable after opening the portal vein， although there was slight increase at prophase in half liver graft transplantation group， and then the portal venous pressure would let down， keeping stable at the later stage. But in small size liver graft transplantation group and extra-small size liver graft transplantation group， the portal venous pressure incr-eased and got the top at 15 minutes after opening the portal vein， and then induced， keeping stable during the 45-60 minutes.Portal venous pressure at the point of 5 （r=-0.942）， 15 （r=-0.947）， 30 （r=-0.900）， 45 （r=-0.825）， and 60 （r=-0.705）minutes after opening the portal vein were significantly related to liver graft size （P＜0.001）. The levels of ALT in wholeliver graft transplantation group and half liver graft transplantation group were both lower than those of small size livergraft transplantation group and extra-small size liver graft transplantation group （P＜0.05）， and levels of ALT in small size liver graft transplantation group was lower than extra-small size liver graft transplantation group too （P＜0.05）. Levelof ALT at 24 hours after transplantation were significantly related to liver graft size （r=-0.685， P＜0.001）. Conclusions The minimum graft volume/standard liver volume （GV/SLV） in reduced size liver transplantation in rat is 50%. The liver graft whose GV/SLV is 30%-35% should be considered as marginal size liver graft， and the liver graft whose GV/SLV less than 30% should be considered as extra-small size liver graft in the rat.
Objective To establish and evaluate the hind limb ischemia model in type 2 diabetic rats， and to providea platform for subsequent intervention experiment. Methods Fifteen SD rats were randomly divided into 3 groups：normal control group， diabetes group， and diabetic ischemia group， each group enrolled 5 rats. The 10 rats of later 2 groups were fed with high fat diet for 4 weeks， and then received intraperitoneal injection of streptozotocin （40 mg/kg） to establish type 2 diabetic model. Three days later， the rats of diabetic ischemia group underwent ligation of the bilateral common iliac artery to establish the hind limb ischemia model， but iliac artery of rats in other 2 groups didn’t received ligation. Ultrasound and color Doppler flow imaging detection was used to determine the peak velocity and acceleration time of femoral artery in rats of 3 groups in 2 weeks after operation， and triceps tissues and quadriceps tissues were collected to perform HE staining and SP staining for the observation on status of nutrition and vascular regeneration respectively. Results The peak velocity of femoral artery in rats of normal control group， diabetes group， and diabetic ischemia group were （22.49±3.02） cm/s， （17.36±2.60） cm/s， and （11.23±1.26） cm/s， the acceleration time were （0.080±0.009） s，（0.120±0.009）s， and（0.160±0.020） s， the arteriolar density measured by immunohistochemistry SP method were 6.80±0.84/HPF， 4.60±0.55/HPF， and 1.40±0.55/HPF respectively. The peak velocity of femoral artery and arteriolar density of rats in diabetic ischemia group were both lower （P＜0.05）， but acceleration time was longer （P＜0.05）. Results of HE staining showed that structure of triceps tissues was damaged， with infiltration of lots of inflamm-atory cells， which was worse than normal control group and diabetes group. Conclusion Method of high-fat diet in combination with small dose of streptozotocin can induce type 2 diabetic rat model， and hind limb ischemia model can be successfully established by ligation of common iliac artery on this model.
Objective To establish interstitial cells of Cajal （ICC） loss models caused by incomplete small intestinal obstruction in rats with modified method and verify it. Methods Modified method was used to establish the models， making the ring around the small intestine but not through it. Morphological changes were observed by general signs， pathological changes were tested by HE staining and transmission electron microscope （TEM）， and changes of ICC number were tested by immunohistochemistry staining. Results Success rate of this method was 56% （28/50）， weight loss happened compared with before operation in ileus group （P＜0.01）. Hyperemia and swelling were observed in ileus group， and gastric retention was obvious. Results of HE staining and TEM showed that there was obvious inflammatory change， and ICC reduced was observed by immunohistochemistry. Conclusion ICC loss models caused by incomplete small intestinal obstruction meet the basic performance， and can be used for further research.
Objective To establish a stable and reliable lung injury model caused by severe acute pancreatitis（SAP）in rats， which is helpful to study the acute lung injury （ALI）and acute respiratory distress syndrome （ARDS） induced by SAP.Methods Sixty Sprague-Dawley rats were randomized into ligature group （n=20）， traditional group （n=20），and sham operation group （n=20）. SAP model was established through retrograde injection of 5% taurocholic acid. After injection， the pancreatic duct of rats was ligated in ligature group， but not in traditional group. The lung damage and edema at 24 h after operaton and natural course of rats were observed.Results The ALI model of rats induced by SAP was established successfully in ligature group. The rats died of acute respiratory failure within 48 h in ligature group， the mortality was significantly higher than that in traditional group （100% vs.20%），P＜0.05. Pleural effusion occurred in four rats in ligature group， while no pleural effusion was found in rats in other two groups. The volume of ascites of rats in ligature group was （21.15±5.33） ml， which was more than that in traditional group 〔（7.75±2.66） ml〕，P＜0.05， while no ascites was found in rats in sham operation group. The level of serum amylase of rats in ligature group was （2 470.70±399.73） U/L，which was significantly higher than that in traditional group 〔（1 528.40±289.54） U/L〕 and sham operation group 〔（831.10±93.26） U/L〕，P＜0.001. The level of serum albumin of rats in ligature group was （6.90±1.66）g/L， which was significantly lower than that in traditional group 〔（13.10±0.99） g/L〕 and sham operation group 〔（16.20±0.92） g/L〕，P＜0.001.The lung wet-to-dry weight ratio （W/D） of rats in ligature group was 6.50±0.23， which was greater than that in traditional group （4.92±0.18） and sham operation group （4.61±0.16）， P＜0.001. The score of lung histopathologic of rats in ligature group was 29.25±1.07， which was significantly higher than that in traditional group （12.65±1.98） and sham operation group （0），P＜0.001. The score of pancreas histopathologic of rats in ligature group was 15.95±0.15，which was significantly higher than that in traditional group （13.75±0.66） and sham operation group （0.13±0.29），P＜0.001. Under transmission electron microscope， basement membrane of pulmonary capillary of rats in ligation group was destructive， the nuclei was dissolved， endothelial pinocytotic vesicles was functional active， and tight junctions between capillary endothelial cells were blurred and even ruptured. Moreover， tight junctions between alveolar epithelial cells were destructive. Pathological changes of lung ultrastructure of rats in ligation group were more severe than that in traditional group， while no pathological change of lung ultrastructure was observed in rats in sham operation group. Conclusions Injury process and pathogenesis of ALI or ARDS clinically caused by acute gallstone pancreatitis can be reproduced in this animal model， which is suitable to explore the related mechanisms of ALI caused by SAP and provides good animal model for the study of ALI caused by SAP.
Objective To establish chronic hindlimb ischemia model with suture-occluded method in rats， and then compare the effects of chronic hindlimb ischemia model with acute ischemia model. Methods Models of chronic hindlimb ischemia were established by using suture-occluded femoral artery method. The laser Doppler blood flow analysis and angiography were performed on day 7， 14， 28， 42， and 49 after operation， and then the rats were sacrificed after angiography， respectively， the quadriceps and gastrocnemius of contralateral and ipsilateral （surgical side） were gotten， which were tested by HE staining and α-actin immunohistochemistry staining， and then calculate arteriolar density. Results There were no lameness and limb necrosis after operation in chronic hindlimb ischemia models. Laser Doppler analysis found that chronic hindlimb ischemia models were still maintained in ischemia state on day 49 after operation compared with acute ischemic models. The resluts of HE staining showed no acute necrosis and muscle fibrosis in chronic hindlimb ischemia model group. Chronic hindlimb ischemia models after operation did not appear obvious lameness and limb necrosis. The arteriolar density of quadriceps femoris on day 7 after operation in chronic hindlimb ischemia models were less than that in acute hindlimb ischemia models （0.015 2 vs. 0.036 4）. Conclusions Compared with the commonly used acute ischemic models， the duration of arterial limb ischemia in chronic hindlimb ischemia rats， which were established by suture-occluded method， is longer and less likely to be affected by the compensatory mechanisms. So suture-occluded method can provide a new animal hindlimb ischemia model for further study of ischemia angiogenesis mechanism and treatment of severe lower extremity ischemia.