Humans ; Infant, Newborn ; Intestinal Perforation ; complications ; diagnosis ; Mycoses ; complications ; diagnosis

Objective To evaluate the effect of pituitrin and nursing for patients with vasoplegic syndrome after cardiopulmonary bypass. Methods 38 patients with cardiopulmonary bypass after open heart operation were divided into two groups with 19 cases in each group according to odd-nunbered and even-numbered days, the PT group was given pituitrin treatment, the NE group was given norepinephrine treatment. Heart rate, MAP, urine volume were recorded at 0.5, 1, 2, 6, 12 and 24 hours after drug therapy.Mechanical ventilation time, ICU stay time,hepatic and renal function and clinical symptom were compared between the two groups. Results MAP and urine volume increased significantly at 0.5 hour after pituitrin therapy. MAP increased from (55.33±4.10)mmHg to (67.34±4.13)mmHg. Urine volume increased from (0.08±0.02)ml·kg-1·h-1 to (1.84±1.21)ml·kg-1 ·h-1 MAP and urine volume also increased at other time points. In the NE group, MAP increased at 1h after treatment. Both MAP and urine volume increased at 2h and later after norepinephrine treatment. One death happened in the NE group. Compared with the NE group, the PT group was associated with lower ICU stay, mechanical ventilation time and 24h serum LAC.Liver and kidney function, myocardial infarction, nausea and vomiting, and other side effects were not different between the two groups. Conclusions Application of low-dosage pituitrin is rapid, efficient and safe treatment for patients with vasoplegic syndrome after open heart operation.

AIM:To explore the inhibitory action of recombinant human tumor necrosis factor-related apoptosis-inducing ligand(rhTRAIL) on mouse breast cancer. METHODS:Each mouse was inoculated 0.2 mL (1?106) D2F2 cells subcutaneously in the right lower limb and they were divided into five groups randomly. The control group was infused PBS 0.2 mL,while the low-dose,medium,high groups received purified rhTRAIL 2.5 mg/kg,5.0 mg/kg,10.0 mg/kg,respectively,the positive group was administered cyclophosphamide 30.0 mg/kg. Every group was operated by peritoneal injection once a day for fifteen days. The mice were weighed every day. The growth state was viewed and the size of the tumor was measured every 3 d to calculate the tumor volume and tumor suppression rate. All mice were killed after 15 d. The pathologic changes of the tumor were observed under light-microscopy and electronic microscopy. The cell cycle and apoptosis index of D2F2 cells were analyzed by flow cytometry. RESULTS:The body weight and tumor volume in low-dose,medium,high groups were lower than those in control group and the restriction effect was more significant than that in the control group (P

BACKGROUND: Skin transplantation is the most effective conventional method to cure large area full-thickness skin damage caused by empyrosis or some diseases, but present deficiency of skin donator is the largest barrier in front. The most ideal way to solve this problem is to use tissue-engineering skin reconstructed by self-skin cells as seed cells.OBJECTIVE: To investigate the effects of tissue engineered artificial animal skin in animal grafting experiment.DESIGN: A randomized controlled trial SETTING: Institute of frontier medical sciences and department of dermatology in a university.MATERIALS: Study was performed in the Cell-Engineering Institute of Jilin University from September 1998 to July 2001. Totally 20 newborn Wistar rats and 24 8-week old male nude mice were selected into our study.METHODS: Full-thickness wounds(diameter: 20 nn) were made on the backs of twenty-four nude mice to establish full-thickness skin defect animal model for grafting by the tissue-engineered reconstructive artificial skin made by chitosan(CH) as stromal scaffold. Twenty-four 8-week old nude mice were divided into artificial skin (AS) group, chitosan membrane(CH) group and control group (CG). All wounds were covered with AS, CH or petrolatun gauze correspondingly. The wounds of each groups were observed daily,techniques like histology and infrared-ray scan were used for a dynamical surveillance on the 3rd, 7th, 14th and 21st days.MAIN OUTCOME MEASURES: ① general observation; ② blood supply in recipient area under infrared-ray observation; ③ histological observation.RESULTS: Transplanted AS had a favorable fusion between tissue-engineered skin and self-skin on the 3rd day after grafting with a few of ingrowths of capillaries. The color of the AS was closed to self-skin. The capillaries gradually increased in the grafts over time. The new epidermis was clearly consisted of stratum basale, stratum spinosum, stratum granulosum, and stratum corneum. Keratinization enhanced with exfoliation. Cells in dermis increased and the scaffold gradually degraded. The secreted extracellular matrix increased as well. On the 14th day after grafting, the wounds almost recovered.The color of the grafted artificial skin was more similar to the nature skin with very little scaring, which indicated that a second grafting was unnecessary. The scabs did not completely fall off in CH group until the 14th day, and the wound was not healed. The color was darker than that of AS group. The scabs fell off in CG, and the wounds were big and deep with dark red color.CONCLUSION: The new reconstructive tissue-engineered artificial skin with CH as stromal scaffold has good histocompatibility, which can be applied in grafting for full-thickness wounds.

Objective :To study a simplified method of isolation of rat hepatocytes and to observe the pro-cess of cell morphology in long-term culture. Methods :Rat hepatocytes were isolated by a single two-stepperfusion method. The yield and viability were assessed by trypan blue exclusion. [3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide] (MTT) was used to test the effect of serum concentration of newborn calf serum on the proliferation of hepatocytes. Hepatocytes were inoculated in the culture mediumconsisted of Williams' E supplemented with insulin,dexamethasone and 10％ new born calf serum. Themorphologic change of cultured hepatocytes was observed. Results:The average yield of hepatocytes was 2.26× 108 cells per rat, with an average viability of 95.6％. New born calf serum had strong biological activi-ty to stimulate the proliferation of hepatocytes and there was close-effect relationship followed by the in-crease of new born calf serum concentration. The rat hepatocytes can be cultured for 5～ 6 weeks withpreservation of normal morphologic appearance. Conclusion:The rat hepatocytes isolated by the abovemethod have high yields and viability and can be long-term cultured in vitro.

OBJECTIVETo study the expression and significance of apoptosis-related protein p53, Bcl-2, and Bax during the development of oral squamous cell carcinoma (SCC).

METHODSThe expression was observed in 10 normal oral epithelia, 48 dysplasia epithelia and 42 SCC by immunohistochemical evaluation.

RESULTSIn normal mucosa, the positive rate of p53, Bcl-2 and Bax were 0%, 20% and 60%. In dysplasia epithelia, the positive rate of p53 is increased (P < 0.05), the positive rate of Bcl-2 and Bax remained no significant change (P > 0.05), but the positive intensity in severe dysplasia was higher than in mild group. In SCC, the positive rate of Bcl-2 increased significantly (compared with dysplasia, P < 0.05), while the expression of Bax was decreased with the increase of SCC histological grade. Further analysis showed the correlation was evident in p53 and Bax in dysplasia, and in p53 and Bcl-2 in SCC.

CONCLUSIONSIn dysplasia, p53 gene mutation results in accumulation of dysplasia cells. In SCC, the cooperation of p53, Bcl-2 and Bax results in the progression of SCC. Apoptosis genes could work either independently or cooperatively.

Apoptosis ; genetics ; Carcinoma, Squamous Cell ; chemistry ; Humans ; Immunohistochemistry ; Mouth Mucosa ; chemistry ; pathology ; Mouth Neoplasms ; chemistry ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Tumor Suppressor Protein p53 ; analysis ; bcl-2-Associated X Protein

Objective To construct the engineering bacteria expressing the recombinant human Kunitz protease inhibitor domain of amyloid protein precursor variant (rhKD/APPvar)in Pichia pastoris,and to establish the methods suitable for large-scale fermentation and purification of rhKD/APPvar.Methods The rhKD/APPvar expression vector was constructed based on the rhKD/APPvar-pPICZαexpression vector. Two restriction enzyme loci (ApaⅠ and SacⅡ)were added to two flanks of KD/APP and human KD/APP activity center RAM was replaced by the active site of BPTI KAR.After the rhKD/APPvar-pPICZαexpression vector was transformed into Pichiapastoris,optimized expression and purification of rhKD/APPvar was performed.The rhKD/APPvar was purified with cation exchange chromatography and desalting.Results The results of digestion identification and DNA sequencing analysis demonstrated that the recombinant plasmid rhKD/APPvar-pPICZα was successfully constructed and transfected into pastoris X-33. The SDS-PAGE analysis results indicated that rhKD/APPvar expressed after the induction of methanol and the relative molecular weight was 6 700.After a series of experiments the optimal expression conditions of rhKD/APPvar were obtained as follows:the optimal pH was 6.0 and the optimal induction time point was about the 5 th day for the strain.After purified the purity of rhKD/APPvar was about 95%.Conclusion KD/APPvar-pPICZ is successfully constructed;after expression in Pichia pastoris and purification,the rhKD/APPvar protein is obtained.

Objective To study the clinical features and follow-up of newborns with severe hypoxic-ischemic encephalopathy ( HIE) , and to provide the basis for rational diagnosis, treatment and follow-up.Methods Clinical data of cases of HIE from the Neonatal Department of our Hospital from January 2011 to October 2014 were studied retrospectively. The data of general information, laboratory examination, treatment, outcome, follow-up and prognosis of the patients were collected. Multivariate logistic regression analysis was used to study the influential factors of the prognosis of HIE.Results A total of 123 infants with sever HIE were enrolled in our study. In addition to general therapy, 6 cases were treated with mild hypothermia, and 21 cases were treated with high pressure oxygen. 60 cases improved our treatment, 55 cases had withdrawal treatment with parental consent, and 8 cases died. Single factor analysis showed that 5 minutes Apgar score, convulsions, coma, pH, BE, organ injury, and mild hypothermia treatment were the risk factors that affect the prognosis of severe HIE. Multiple factors analysis showed that 5 min Apgar score <3 points ( OR=4. 071 ,95℅CI 1. 309-15. 613 ) and BE≤-10 mmol/L ( OR=36. 810, 95℅CI 5. 913-41. 119) were independent risk factors of prognosis of severe HIE ( P<0. 05). Hospitalization within the first 72 hours of life ( OR=0. 096, 95℅CI 0. 096-0. 353) was a protective factor of severe HIE. Multiorgan injury ( mainly the injury of brain, lung and heart) and electrolyte imbalance ( mainly hypocalcemia and hyponatremia ) were common complications of serve HIE. In the follow-up of these patients, 33 cases were loss in follow up, and 49 cases died (8 cases died during hospitalization, 41 cases died after withdrawal of treatment). The top five causes of death were abandonment of treatment due to financial reasons and the fear of adverse outcome (n=20), multiple organ dysfunction ( n =16 ) , and pneumothorax ( n =4 ) , diffuse intravascular coagulation (n=6), and shock (n=3). 41 cases survived were followed up for 9~54 months. The critical clinical conditions observed among these infants included cerebral palsy ( n = 5 ) , epilepsy ( n = 3 ) and developmental retardation(n=26).Conclusions There are many complications of severe HIE.The mortality of severe HIE is high, and the incidence of poor outcome of survivors is also high. Timely detection of risk factors is the key to the prevention of severe HIE. Long-term prognosis of severe HIE requires proper organization of neonatal follow up.

Objective Toinvestigatetheeffectofrehabilitationrobothandontheupperlimbmotor functioninpatientswithstrokeandhemiplegia.Methods Theeffectsoftraditionalrehabilitation therapy or robot rehabilitation treatment of 15 stroke patients with hemiplegia were analyzed retrospectively. There were 4 females and 11 males. According to the group order,they were divided into either a robot rehabilitation group (n=8 )or a general rehabilitation group (n =7 ). The patients of the general rehabilitation group received general rehabilitation training at the Department of Rehabilitation Medicine for 4 weeks,3 times a week,and 30 min for each time. At the same time,they also conducted family rehabilitation training,5 times a week,once for 1 h;the patients of the robot rehabilitation group received the general rehabilitation training for the same length of time as the general rehabilitation group,and they also performed robot rehabilitation training,5 times a week,once for 1 h. The patients of both groups were evaluated with the modified Ashworth scale (MAS )scores,Fugl-Meyer motor assessment of the upper extremity(FMA-UE),andWolfmotorfunctiontest(WMFT)beforeandaftertreatment.Results Compared with before training,the MAS score of the robot rehabilitation group was decreased significantly after four weeks of training. The time in WMFT was shortened significantly and the function score was improved obviously. The scores of upper extremity,hand score,and total FMA score in FMA-UE were improved significantly,and there were significant differences (all P <0. 05 ),while there were no significant differences in various indicators in the general rehabilitation group (all P>0. 05). Comparing the effects after four weeks of rehabilitation in both groups,the robot rehabilitation group was superior to the general rehabilitationgroupforthehandscoreinFMA-UE(allP<0.05).Conclusion Therehabilitation robot hand effectively lowered the upper limb spasticity. It has significant effect for recovery motor function of hand;however,it did not have a significant effect for motor rehabilitation on wrist.

Objective To investigate the effect of illness severity on preterm infant's hypothalamusputituary-adrenal (HPA) axis, we measured the serum concentration of cortisol,aldosterone and adrenocorticotropic hormone (ACTH). Methods Ninety preterm infants who were transferred to our hospital within 72 hours after birth were involved. These preterm infants were divided into two groups:gestational age (GA) ≥34 weeks' preterm infants and GA <34 weeks' preterm infants. We evaluated these preterm infants at the time of admission,day 7 and day 14 after birth with neonatal critical illness score (NCIS). Then they were divided into mild group and severe group by the lowest score. We measured their serum cortisol,aldosterone and ACTH at the time of admission,day 7 and day l4 after birth. Results (1) The serum cortisol concentration of preterm infants with severe illness was higher than that of preterm infants with mild illness. Among the GA ≥34 weeks' preterm infants,the serum cortisol concentration of preterm infants with severe illness was significandy higher than that of preterm infants with mild illness within 72 hours after birth (t = -2.263,P =0. 029). Among the GA <34 weeks' preterm infants,the serum cortisol concentration of preterm infants with severe illness was significantly higher than that of preterm infants with mild illness on day 14 after birth (t =-2. 913 ,P =0. 006). (2) Among the preterm infants with severe illness,the serum cortisol concentration of the GA≥34 weeks' was significantly higher than that of the GA < 34 weeks' within 72 hours after birth (t =-2. 641 ,P =0. 010) ;the serum cortisol concentration of the GA <34 weeks' was significantly higher than that of the GA≥34 weeks' on the day 14 after birth(t = -2. 189,P =0. 036) . (3) The serum cortisol concentration was significantly decreased in the GA≥34 weeks'preterm infants (F = 4. 679, P =0. 012). (4) The serum cortisol concentration of aldosterone and ACTH was not significantly different between preterm infants with severe illness and those with mild illness. Conclusion The preterm infant already has the ability to respond to stimuli by regulating cortisol secretion. The serum cortisol concentration increases as disease severity worsens.Serum aldosterone and ACTH concentration are not correlated with the severity of the disease.