BACKGROUND: To prepare a novel biological material that can be implanted into the lesion of bone tuberculosis, keep sustained release of anti-tuberculosis drugs around bone tuberculosis tissues for a long time, and enhance the effect on bone repair has become a hot spot for clinical studies on bone tuberculosis.OBJECTIVE: To prepare the adipose-derived mesenchymal stem cells (ADMSCs)/sustained-release rifampin-loaded microsphere complex, and to preliminarily study its effects in a rabbit model of spinal tuberculosis.METHODS: Rifampin-chitosan-calcium alginate sustained-release microspheres were synthesized, and co-cultured with ADMSCs to prepare an anti-tuberculosis composite for bone tissue engineering. Forty New Zealand rabbits were used to make lumbar L6 tuberculosis models. Thereafter, rat models were randomly divided into four groups and given rifampin by gavage in rifampin group, ADMSCs by paravertebral injection+rifampin by gavage in stem cell group, rifampin by gavage+implantation of the anti-tuberculosis composite material in experimental group, and no treatment in control group.The duration time of rifampin administration was 8 weeks. The anti-tuberculosis effect of the composite material was evaluated by X-ray and CT scanning observation.RESULTS AND CONCLUSION: In the control group, obvious damage to lumbar vertebrae L5 and L6 was apparent; inflammatory granulation tissues formed; and the intervertebral space was narrowed. In addition, two rabbits in the control group showed obvious kyphotic deformity and five showed pasoas major swelling with low-density dark region in the psoas muscle. In the rifampin group, there were five rabbits with moderate damage of the lumbar vertebrae L5 and L6,and two rabbits with pasoas major swelling. In the stem cell group, there were two rabbits with moderate damage of the lumbar vertebrae L5 and L6, three rabbits with mild damage of the upper part of the lumbar vertebra L6, and three rabbits with pasoas major swelling. In the experimental group, only four rabbits suffered from mild damage of the upper part of the lumbar vertebra L6 but with no changes in the intervertebral space between the L5 and L6, and without pasoas major swelling. These results indicate that the composite material of ADMSCs combined with sustained-release rifampin-loaded microspheres can inhibit mycobacterial growth effectively, and reduce vertebral bone destruction,thereby giving some therapeutic actions for the animal models with spinal tuberculosis.

Objective To analyze the clinical features of the hyperamylasemia in critically ill children and investigate the related risk factors in order to provide the basis for prevention and treatment. Methods A total of 1036 critically ill children admitted in pediatric intensive care unit (PICU)from April,2011 to Oct,2012 were studied.They were divided into the high amylase group (n=82)and the normal group (n=954).According to the outcomes,the high amylase group was divided into survival group (n=61 ) and death group (n =21 ).The related risk factors of the occurrence and outcome of hyperamylasemia were analyzed by univariate and multivariate Logistic regression.Results There were statistically significant differences in rates of coagulation disorders, convulsions, disturbance of consciousness,pediatric critical illness score (PCIS)≤80,multiple organ dysfunction (MODS)≥3, sepsis,shock,and lactic acid (LA),procalcitonin (PCT),blood glucose (BG)between the high amylase group and the normal group (P<0.05 ).The differences in the rates of coagulation disorders,convulsions, mechanical ventilation,PCIS≤80,MODS≥3,and LA,PCT,oxygenation index,albumin,C-peptide,BG were statistically significant between the survival group and the death group (P <0.05 ).Multivariate Logistic regression analysis showed that the risk factors of the hyperamylasemia's occurrence were LA,PCT, BG,PCIS<80,MODS>3.Adjusted ORs confidence intervals of them were 1.662 (1.236-2.234),1.042 (1.025-1.060),1.612 (1.411-1.843),3.219 (1.311-7.905),3.411 (1.370-8.494),respectively. The hyperamylasemia's prognostic risk factors were PCT,C-peptide,PCIS ≤80,MODS >3,shock. Adjusted ORs confidence intervals of them were 1.066(1.021-1.113),1.437(1.017 ~2.030),16.137 (1.876-138.836),10.437(1.528-71.925),20.928(1.938-226.009),respectively.Conclusions The severity of the disease,the levels of LA,PCT,BG in critically ill children were positively correlated to the occurrence of hyperamylasemia.The severity of the disease,the incidence of organ failures,the levels of PCT,C-peptide combined shock in children with hyperamylasemia were positively correlated to the prognosis of hyperamylasemia.

Two new sesquiterpenoids, named as tyromols A and B (1 and 2), were isolated from cultures of basidiomycete Tyromyces chioneus, along with two previously reported 15-hydroxy-6 α, 12-epoxy-7β, 10αH, 11βH-spiroax-4-ene (3) and agripilol C (4). Compounds 1-4 were separated and purified by silica gel, RP-18, Sephadex LH-20 column chromatography. Their structures were elucidated on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments.

Objective To analyze the clinical features of critically ill children with abnormal pancreatic ultrasound findings in order to explore the related risk factors for offering evidence-based diagnosis of pancreatic damage secondary to critical illness.Methods A prospective study was performed in 531 critically ill children admitted to pediatric intensive care unit(PICU) of 17 children' hospitals from January 2012 to March 2014.All patients were divided into control group(513 cases) and abnormal group(18 cases) according to the pancreatic ultrasound findings.Comparison of clinical features and biochemical indicators were made between two groups.The related risk factors associated with abnormal pancreatic ultrasound findings were analyzed by using Logistic regression analysis.ROC curves were used to evaluate the role of amylase and lipase in the diagnose of abnormal pancreatic ultrasound findings.Results The incidence of abnormal pancreatic ultrasound findings in critically ill children was 3.39%, the average age of abnormal group was significantly older than that in control group (P<0.01).There were not statistically significant differences in gender and primary disease between two groups.The incidence of hypotension was 22.2% and the incidence of abdominal muscle tension was 16.7% in abnormal group,which were statistically higher than those in control group (P<0.05).The levels of calcium, albumin of abnormal group were significantly lower than those in control group (P<0.01), and levels of serum amylase, lipase, lactate dehydrogenase in abnormal group were significantly higher than those in control group (P<0.01).The risk factors associated with abnormal pancreatic ultrasound findings were age, blood pressure, calcium,amylase.Area under the ROC curve of abnormal pancreatic ultrasound findings determined by amylase and lipase were 0.803 and 0.745,respectively (P<0.05).The sensitivity was 0.667, specificity was 0.881 when the serum amylase was 101.5 U/L, the sensitivity was 0.722, specificity was 0.928 when the serum lipase was 96.9 U/L.Conclusions The incidence of abnormal pancreatic ultrasound findings was rather low in critically ill children.The risk factors associated with abnormal pancreatic ultrasound findings were hypotension, hypocalcemia, and hyperamylasemia.The elevated serum amylase and lipase might be the most likely factors associtaed with abnormal pancreatic ultrasound findings.

Objective To discuss the blockage reasons and countermeasures of temporary catheter in central venous including femoral vein and internal jugular vein for hemodialysis patients.Methods The reasons of blockage of temporary catheter in central venous among 60 hemodialysis patients from January 2014 to September 2015 in Hemodialysis Room of the First People's Hospital of Longquanyi district of Chengdu were reviewed.There were 32 cases in femoral vein and 28 cases in the right internal jugular vein.The incidence of blockage of the two temporary catheter in the first two weeks,the incidence of blockage caused by mechanical factors and blood clots or fibrous sheath and the incidence of blockage which needed to change the catheter in the two groups patients within the first two months were analyzed.Results The first rate of defective function in the first two weeks of femoral vein group was 53.13％(17/32),and 7.14％(2/28) of the right internal jugular vein group,there were significant difference between the two groups (x2=13.061,P＜0.001).The incidence of catheter blockage due to mechanical factors of femoral vein group was 96.88％ (31/32),and 53.57％ (15/28) of the right internal jugular vein,there were significant difference between the two groups (x2 =15.654,P＜0.001).The incidence of blockage caused by blood clots or fibrous sheath and the incidence of blockage which needed to change the catheter in the first two months were no significant differences between the two groups(P=0.061,0.096).Conclusion The cases of temporary catheter in femoral vein have blockage earlier,and they have a higher risk of catheter jam than the cases of temporary catheter in right internal jugular vein.

As an exposing part of the human body,facial surface is easy to be injured in our daily lives.With the increase of living standards,the patients have an ever more urgent require for aesthetic treatment for the medical and plastic surgical treatment for the exposing parts of body,especially for the facial surface.But we found in the clinical work some aesthetic treatment were not satisfying,and some patients even needed a second restitution.We have analyzed the reasons for this phenomenon and proposed some corresponding solving methods.

Objective To observe the effect of the nutrition management according to the JCI(Joint Commission on Accreditation of Healthcare Organizations)management standard for critically ill pediatric patients with mechanical ventilation.Methods A total of 330 cases with mechanical ventilation were en-rolled in the control group from Hunan Province Children's Hospital PICU between Jan.2012 and Dec.2012, and these pediatric patients were managed with the conventional nutrient management.A total of 359 pediatric patients with mechanical ventilation were admitted to the experimental group from Jan.2013 to Dec.2013 and these patients were managed with nutrient management based on JCI standards.The length of mechanical venti-lation,stay in ICU,stay in hospital were compared between two groups,and the incidence of ventilator-associ-ated pneumonia,abandonment rate and mortality were also compared between experimental group and control group.Results The length of mechanical ventilation in control group[(8.39 ±1.34)days]was longer than that of experimental group[(5.69 ±1.12)days].The length of stay in PICU for control group[(12.32 ± 1.37)days]was more than that in experimental group[(9.42 ±1.53)days].The length of stay in hospital for control group [(15.37 ±2.16 )days]was higher than that of experimental group [(12.63 ±2.29 ) days].The incidence of ventilator-associated pneumonia(9.7%)in control group was higher than that of ex-perimental group,and the giving up or mortality rate in control group(8.48%)was higher than that of exper-imental group(4.35%).And there were significant differences by statistical analysis(P ﹤0.01 ,respective-ly).Conclusion According to the nutritional management in JCI standard,the length of mechanical ventila-tion,stay in PICU and stay in hospital time,the incidence of ventilator-associated pneumonia,and the aban-doned or mortality rate were reduced for critically ill pediatric patients with mechanical ventilation.

Objective To analyse the relationship between insulin resistance and the pancreatic beta cell dysfunction in critically ill children with hyperglycemia,to investigate the relationship between the pancreas injury and pancreatic beta cells dysfunction.Methods Seven hundred and thirty-six critically ill children admitted in PICU of Hunan Children's Hospital from Nov 2012 to Mar 2013 were reviewed and analyzed.According to the maximum intravenous blood glucose within 24 h after admission,they were divided into severely elevated group (blood glucose ≥11.1 mmol/L,n =67),slightly elevated group(blood glucose 6.1 ～ 11.1 mmol/L,n =361) and control group (blood glucose ≤ 6.1 mmo/L,n =308).Serum insulin,C peptide,serum amylase,lipase,urinary amylase,HOMA-β,HOMA-IR were compared among 3 groups.According to the severity of sepsis,they were divided into non-sepsis group (n =414),sepsis group (n =237),severe sepsis group (n =64),septic shock group (n =21).Blood glucose,serum insulin,C peptide,HOMA-βand HOMA-IR were compared among 4 groups.Results (l)The levels of insulin,C peptide,blood amylase,lipase and urine amylase were gradually increased with elevated blood sugar(rs =0.235,P ＜ 0.05;C =0.142,P ＜ 0.05 ; rs =0.142,P ＜ 0.05 ; rs =0.119,P ＜ 0.05 ; r.s =0.093,P ＜ 0.05).The differences among 3 groups were significant (P ＜ 0.05).The levels of serum amylase (IU/L) [102.81 (10.48-191.69)],lipase(U/L) [69.75(10.67-121.85)] were higher than upper limit of normal in severely elevated group.HOMA-β fell to 18.75％ in severely elevated group.The level of HOMA-β was decreased with the increase of blood sugar level (rs =-0.108,P ＜ 0.05).The level of HOMA-IR was increased with the elevated blood sugar level(rs =0.455,P ＜ 0.05).(2) The levels of blood glucose,insulin,C peptide and HOMA-β were significantly different among 4 groups of non-sepsis,sepsis,severe sepsis and septic shock group (P ＜ 0.05),HOMA-IR showed no significant difference among 4 groups (P ＞ 0.05).The levels of blood sugar increased to 9.21 (6.21-19.60) mmol/L,HOMA-β declined to 10.52％ in septic shock group,and blood glucose,insulin,C peptide,HOMA-β were significantly different compared with the other 3 groups (P ＜ 0.05).Conclusion Hyperglycemia is associated with insulin resistance and pancreatic β cells dysfunction in critically ill children,the cause of beta cell dysfunction is secondary to pancremic injury.Pancreatic beta cells dysfunction inducing hyperglycemia is more significant than insulin resistance in sepsis children.

BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.

The present study is to explore the change process and distribution of phosphorylated DARPP-32 (p-DARPP-32) in rat brain including cortex, hippocampus and striatum and to further deduce whether p-DARPP-32 was possibly involved in epilepsy induced by repetitive low doses of pentylenetetrazol (PTZ). PTZ-induced epilepsy model in rat was established with 30 male SD rats randomly divided into 6 groups, control group and five trial groups [PTZ 1 h, PTZ 6 h, PTZ 24 h, PTZ 48 h and PTZ 72 h respectively, after onset of status epilepticus (SE)]. Immunohistochemistry and immunofluorescence double-labeling were used to detect the temporal time change and distribution of p-DARPP-32 expression and to analyze the coexpression of DARPP-32 and p-DARPP-32 in rat brain after the onset of PTZ-induced generalized SE. The results showed that there was a temporal time change of p-DARPP-32 expression in rat brain after the onset of SE. The number of p-DARPP-32-positive cells increased significantly and reached the peaks at the ends of 1 hour and 6 hours after the onset of SE, but decreased at the end of 24 hours. The moderate to strong p-DARPP-32-immunopositive neurons were observed in cortex, hippocampus and striatum, and located in cell cytoplasm and cell nucleus. Further immunofluorescence double-labeling revealed that denser colocalization of p-DARPP-32 and DARPP-32 in the neurons existed in the area mentioned above. Therefore, PTZ-induced SE may cause phosphorylation of DARPP-32 in rat brain. The temporal time change and distribution of p-DARPP-32 suggest that phosphorylation of DARPP-32 may be involved in PTZ-induced epilepsy in rat brain including cortex, hippocampus and striatum, and p-DARPP-32 may play a central role in the onset of SE.
Animals ; Cerebral Cortex ; metabolism ; Corpus Striatum ; metabolism ; Dopamine and cAMP-Regulated Phosphoprotein 32 ; metabolism ; Hippocampus ; metabolism ; Male ; Neurons ; metabolism ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; metabolism