Volume 86  ·   Issue 13  ·  2006

Objective to evaluate the intervention effect of carvedilol, a new generation beta receptor blocker, on cardiomyocyte apoptosis and related gene expression after acute myocardial infarction (AMI) in rats.Methods Eighty-three female SD rats who survived 24 h after coronary artery ligation were divided into AMI control (MI group, 43 rats) and carvedilol (10 mg · kg-1 · d-1) treatment group (C group, 40 rats), and sham operation group (S group, 27 rats). Each group was divided into two subgroups according to the observation time point: 48 h and 4 weeks. Group C was administered by direct gavage 24 h after surgery. The apoptosis of cardiomyocyte was detected by TUNEL method and DNA gel electrophoresis. The expression of "apoptosis inhibiting complex gene" bcl-2/bax was detected by immunohistochemistry and Western blot method. Results (1) The apoptosis index of cardiomyocyte in infarct/scar area, infarct margin area and non-infarct area in MI group (21.2%±15.6%, 19.8%±12.0% and 6.3%±4.5% at 48 h; 23.5%±13.0%, 8.0%±4.4% and 3.2%±1.6% at 4 weeks) was higher than that in prosthetic hand<0.05); The expression of apoptosis-promoting gene bax was significantly higher in MI48H group than that in sham-operated group, but the expression of "apoptosis-inhibiting gene" bcl-2 increased only in infarcted myocardial cells in MI48H group; The ratio of bcl-2/bax was lower in the MI48 h group (1.06) than that in the sham operation group (1.87). (2) There was no significant difference in the apoptosis index of myocardial cells in the three regions of the C48 h group compared with the MI group (all P>0.05), but the expression of bcl-2 in the infarct area and the marginal area was increased; The apoptosis indexes of myocardial cells in infarct/scar area and marginal area in C4-week group (4.0%±2.0%,2.9%±1.6%) were lower than those in MI group (23.5%±13.0%,8.0%±4.4%, all P<0.05), and the expression of bax in cardiomyocytes in scar area was also significantly decreased, but the expression of bcl-2 was not significantly changed; The ratio of bcl-2/bax in myocardial cells of rats in group C at 48 h and 4 weeks (1.72, 2.23) was higher than that in MI group (1.06, 1.8), and was equivalent to that in sham operation group (1.87, 2.25). Conclusion After AMI occurred in rats, carvedilol treatment for a long time (4 weeks) can effectively prevent the apoptosis of myocardial cells in infarct/scar area and its marginal area, and increase the ratio of bcl-2/bax.

Objective to investigate the effect of human HO-1 gene transfection with adenovirus vector on adult pancreatic islets cultured in vitro, and to explore the potential application value of gene therapy in pancreatic islet transplantation.Methods Adult pancreatic islets were isolated and purified and divided into three groups: transfected with human HO-1 genome (Ad-HO-1 group), transfected with EGFP genome (Ad-EGFP group) and control group.Adult pancreatic islets cultured in vitro were transfected with adenovirus carrying human HO-1 gene and EGFP gene as vector, and apoptosis was detected by flow cytometry after 48 h induction by tumor necrosis factor (TNF) α and cycloheximide.Results The insulin secretion of pancreatic islets in Ad-HO-1 group was 270 mIU/L ± 89 mIU/L under high glucose stimulation, which was higher than that in control group (182 mIU/L ± 59 mIU/L) and Ad-EGFP group (189 mIU/L ± 88 mIU/L, P<0.05); After induction, the incidence of apoptotic cells in Ad-HO-1 group was 63%±11% and that in control group was 91%±11%.<0.01. Conclusion the transfection of HO-1 gene into adult pancreatic islets cultured in vitro by adenovirus can increase the anti-apoptotic ability of pancreatic islets and promote insulin secretion.

Objective to investigate the effect of sinomenine, an inhibitor of cyclooxygenase 2 (COX-2), on acute heart graft rejection in rats. Methods The rats were randomly divided into experimental group (20 rats) and control group (20 rats) after 40 SD → Wistar abdominal heart transplantation. The rats in the experimental group were given sinomenine 30 mg · kg-1 · d-1, and the control group was given physiological saline. The survival time was evaluated, the transplanted heart sections were examined, and the pathological analysis of rejection reaction and the activity of COX-2 were evaluated. Results The survival time of graft in the experimental group was 12.5 d ± 2.6 d significantly higher than that in the control group (6.8 d ± 0.5 d) (P =0.001). At 3 and 5 d after transplantation, the inflammatory reaction, vascular intimal inflammation, myocardial edema and myocardial necrosis in the experimental group were significantly reduced, and the apoptosis of the transplanted heart was significantly reduced (P<Conclusion Sinomenine can prolong the survival time of graft and alleviate myocardial damage and inflammatory reaction.

Objective to observe the effect of local mild hypothermia on the expression of aquaporin 4 (AQP-4) mRNA and protein in rat model of cerebral hemorrhage by autologous blood injection, and to explore the possible mechanism of local mild hypothermia in reducing edema after cerebral hemorrhage. Methods 240 male Wistar rats were randomly divided into cerebral hemorrhage (ICH) group and cerebral hemorrhage plus local mild hypothermia (ICH + H) group. Each group was divided into 6 subgroups: control, 6, 24, 72 h, 5, 7 d after cerebral hemorrhage. ICH + H group was treated with local mild hypothermia for 4 h after blood injection, and the blood-brain barrier (BBB) permeability and brain water content were detected in each subgroup, and AQP-4 was measured by reverse transcription (RT) -PCR and Western blot. Results The increase of brain water content, BBB permeability and AQP-4 mRNA expression in ICH group started at 6 h after cerebral hemorrhage, and the increase of AQP-4 protein expression started at 24 h after cerebral hemorrhage, and reached a peak at 72 h (P<The changes of AQP-4 mRNA and protein expression were positively correlated with the changes of BBB permeability (r =0.78 and r =0.76). The brain water content and BBB permeability in ICH + H group decreased significantly at each time point compared with ICH group, while the AQP-4 mRNA absorbance (a) was lower at 48 h after cerebral hemorrhage compared with ICH group, and decreased from 1.25±0.03 to 1.04±0.02 at 72 h (P<The value of AQP-4 protein A decreased significantly at each time point compared with that of ICH group, from 0.77±0.08 to 0.25±0.04 in ICH group at 72 h (P<Conclusion the destruction of BBB integrity can lead to the increase of AQP-4 expression after cerebral hemorrhage. Local mild hypothermia can inhibit the increase of AQP-4 mRNA and protein expression after cerebral hemorrhage. Local mild hypothermia may inhibit the increase of AQP-4 mRNA expression at the transcriptional level by reducing the destruction of BBB integrity, or directly inhibit the expression of AQP-4 protein at the translational level, so as to alleviate the formation of cerebral edema after cerebral hemorrhage.

Objective to investigate the effect of cyclin-dependent kinase inhibitor olomoucine on the microenvironment of axonal regeneration after spinal cord injury and its significance. Methods A spinal cord hemisected injury model was established, and the experimental rats were randomly divided into sham operation group, injury control group and olomoucine intervention group. The expression of cell cycle related proteins after spinal cord injury was analyzed by Western blot. The expressions of glial fibrillar acidic protein (GFAP), chondroitin sulfate proteoglycan (CSPG) and growth-associated protein 43 (GAP-43) were detected by immunofluorescence technique. The motor function of paralyzed hind limbs of rats was evaluated by modified Gale combined score. Results the expressions of cyclin A, B1, E, proliferating cell nuclear antigen, GFAP, CSPG and GAP-43 were weak in the sham-operated group; After spinal cord injury, the expressions of cyclinA, cyclinB1, cyclinE and cyclin (PCNA) were significantly increased, astrocytes were activated and proliferated, and the expressions of GFAP and CSPG were significantly enhanced (all P<0.01), the expression of GAP-43 was higher than that of sham-operated group (P<0.05); After olomoucine intervention, the expression of cell cycle-related proteins was significantly downregulated, the proliferation of astrocytes and the formation of glial scars in the injured area were inhibited, the expression of CSPG produced by glial scars was significantly reduced, the expression of GAP-43 was significantly increased, and the motor function of paralyzed hind limbs was significantly improved (all P<Conclusion olomoucine can regulate the cell cycle, inhibit the activation and proliferation of glial cells and the formation of glial scar after spinal cord injury, reduce the expression of inhibitory factor CSPG and upregulate the expression of GAP-43, which is beneficial to axon regeneration, improve the microenvironment of axon regeneration after injury, and finally promote the recovery of motor function of paralyzed hind limbs.

Objective to investigate the role of chemotherapy in the treatment of patients with recurrent endometrial carcinoma. Methods Twenty patients with recurrent endometrial carcinoma were retrospectively studied. The main contents of the analysis were the recurrence of tumor, the treatment after recurrence and the effect of chemotherapy on prognosis. Results 20 patients with recurrence, accounting for 11.0% of the total number of patients with endometrial carcinoma in the same period, recurrence occurred on average (17 ± 10) months after initial treatment. Among the 20 patients with recurrence, 11 (55.0%) received chemotherapy for an average (6 ± 4) course of treatment. After retreatment, the average follow-up time of 20 patients was (17 ± 16) months. Among them, 1 (5.0%) survived without tumor, 4 (20.0%) survived with tumor, and 15 (75.0%) died due to cancer. Univariate analysis showed that whether or not to receive chemotherapy and the number of chemotherapy courses were significantly correlated with the prognosis of patients (all P<0.05), and whether or not to receive radiotherapy was also correlated with the prognosis (P<0.05). However, the pathological type of tumor, the stage of primary operation, the degree of tumor cell differentiation, age and recurrence time, whether or not to receive surgery, different chemotherapy regimens, chemotherapy routes and whether or not to receive endocrine therapy had no correlation with the prognosis of patients Chemotherapy has a significant effect on the prognosis of patients after recurrence (β = -1.349, OR =0.259, P<0.05). Conclusion Comprehensive treatment can improve the survival rate of patients with recurrent endometrial cancer, and chemotherapy plays a leading role in it, which has an important effect on the prognosis of patients.

Objective to provide scientific basis for determining the reference value of ultrasonic bone mineral density and the diagnostic criteria of osteoporosis in healthy people in China. Methods Calcaneal stiffness index was measured in 2769 healthy people (1262 males and 1507 females) with Achilles Express ultrasonic bone mineral densitometry apparatus produced by Lunar Company in the United States. The changes of calcaneal stiffness index with age, height and weight and the prevalence of osteoporosis in healthy people of different genders in Zhejiang Province were investigated. According to different genders, every 5 years old was divided into one age group, and over 64 years old was combined into one group, 12 groups of males and females were respectively analyzed. Results The peak values of calcaneal stiffness index of males and females appeared at 20~24 years old and 15~19 years old, respectively, and then gradually decreased with the increase of age. After 55 years old, the stiffness index of females decreased faster than males, The prevalence of osteoporosis was also higher than that of males. The difference between the hardness index T value obtained by the instrument (using Japanese "young adults" as reference) and the actual T value (using this study population "young adults" as reference) was statistically significant (instrument T value, male different age groups χ 2 test: χ 2=71.049, df =18, P<0.01; Female: χ 2=237.828, df =18, P<0.01; Actual T value, male different age groups: χ 2=69.897, df =18, P<0.01; Female: χ 2=239.324, df =18, P<Conclusion the calcaneal stiffness index is correlated with age, height and weight in both sexes. The distribution of bone mineral density in the reference population of imported instruments is not completely consistent with that of Chinese, and the diagnostic criteria for Chinese osteoporosis need to be determined.

Objective to investigate the feasibility of allogeneic articular chondrocytes as cartilage seed cells in repairing articular cartilage defects? Methods the articular cartilage of limbs of adult New Zealand white rabbits was aseptically collected, the cells were isolated by enzyme digestion, cultured and expanded in vitro in conditioned medium, embedded in calcium alginate gel for 1 week, and implanted into the intercondylar cartilage defect of knee joint of rabbits, and the defect was left in the control group. The samples were collected 3 and 6 months after operation, respectively, to observe the defect repair; Results 7/8 defects were repaired by mature hyaline cartilage tissue and 1/8 by fibrocartilage; Wakitani has an average rating of 1.75; There was no infiltration of lymphocytes or leukocytes in the 3-month group and no obvious degeneration in the 6-month group in the experimental group. Conclusion It is feasible to repair knee cartilage defects by embedding allogeneic articular chondrocytes with alginic acid, and the allogeneic articular chondrocytes can become seed cells for tissue engineering of articular cartilage after proper treatment.

Objective to explore the changes and expression of specific genes in nasal NK/T cell lymphoma (N-NK/T-L). Methods Restriction enzyme digestion pathway genome scanning (RLGS) was performed on N-NK/T-L tumor cells and peripheral blood leukocytes in the same case, combined with the corresponding virtual genome scanning (VGS) bioinformation analysis and gene bank search to determine the abnormal gene signals; By cloning the abnormal genes found in the above results and preparing probes, Southern and dot blot hybridization were performed on 3 fresh N-NK/T-L tumor tissues and 1 peripheral blood sample, and in situ hybridization was performed on N-NK/T-L in 20 paraffin tissues and control group including 17 B cell lymphoma at the same site, 3 normal spleen tissues and 5 chronic inflammatory nasal mucosa tissues. Results RLGS and VGS methods showed that there was abnormal T-bet gene in N-NK/T-L tumor cells; The T-bet gene was amplified in 3 fresh N-NK/T-L tumor cells by Southern and dot blot hybridization; In situ hybridization showed that the positive expression of T-bet gene was found in 18 (90.0%) of 20 cases of N-NK/T-L, but in only 2 (11.8%) of B cell lymphoma. The difference between the two cases was statistically significant (P<Conclusion T-bet gene is amplified and overexpressed in N-NK/T-L tumor cells. The abnormal expression of T-bet gene may play a role in the occurrence of N-NK/T-L, and it is necessary to further study whether it can be used as classification basis and auxiliary diagnostic index.

Objective to investigate the effect of prophylactic application of pulmonary surfactant (PS) on neonatal respiratory distress syndrome (NRDS)<1500 g, gestational age<High-risk premature infants at 32 weeks and within 0.5 h after birth. Clinical and chest radiographic manifestations of no respiratory distress syndrome (RDS) before the prevention of PS. Microbubble stabilization test of gastric juice was performed<The patients with RDS were randomly divided into prevention group and non-prevention group (50 cases in each group). The prevention group was given Gursu immediately after admission, while the non-prevention group was given PS after RDS. Results Compared with those before administration, the arterial partial pressure of oxygen (PaO2), arterial/alveolar partial pressure of oxygen (A/APO2), pH, arterial partial pressure of oxygen/inhaled oxygen concentration (PaO2/FiO2) increased significantly, while the arterial partial pressure of carbon dioxide (PaCO2) decreased significantly (86.2 mm Hg ± 8.1 mm Hg, 0.30 ± 0.04, 7.38 ± 0.06, 178 ± 37, 37.3 mm Hg ± 9.8 mm Hg vs 45.8 mm Hg ± 10.4 mm Hg, 0.12 ± 0.02, 7.18 ± 0.09, 63 ± 16, 53.6 mm Hg ± 11.1 mm Hg, P all<0.01), and the chest X-ray appearance was also significantly improved 6 h after administration (1.89 ± 0.34 vs 3.02 ± 0.23, P<0.01). FiO2 and mean airway pressure (MAP) decreased significantly after treatment (6.0 cm H2O ± 0.3 cmH2O vs 9.0 cm H2O ± 1.0 cm H2O, P<The incidence of RDS in the prevention group was lower than that in the non-prevention group (30% vs 64%, χ 2=6.032, P<0.01), the incidence of severe illness and mortality were also significantly reduced (20% vs 53%, 0% vs 25%, respectively) (χ 2=6.741, P =0.01; χ 2=4.519,<The total mechanical ventilation time, aerobic time and hospital stay of RDS patients in the prevention group were also significantly shorter than those in the non-prevention group (3.6 d ± 1.7 d vs 5.9 d ± 3.6 d, t =4.692, P<0.05; 8.6 d ± 5.5 d vs 14.1 d ± 6.2 d, t =4.35, P<0.01; 20.5 d ± 10.0 d vs. 32.8 d ± 17.8 d, t =2.952, P<Conclusion PS replacement therapy can improve the pulmonary ventilation function of children with NRDS. Prophylactic supplementation of PS in children with high risk of developing NRDS can reduce the incidence, severity, mortality and complications of RDS.

Bronchial asthma is a disease that seriously endangers the health of the masses, affects the social labor force and the quality of life of patients. In recent ten years, the understanding of asthma in modern medicine has made remarkable progress, which can be summarized as follows: (1) It is clear that the essence of bronchial asthma is a chronic allergic airway inflammation, which still exists in the airway wall even in the remission period, and the application of glucocorticoid can effectively control the airway inflammation of asthma patients; (2) The inhalation technique is the main drug administration method, because this method has rapid onset of medication, convenient medication and few adverse reactions, thus making it possible to use glucocorticoids for a long time; (3) The treatment and prevention of asthma is a long-term task, and doctors need to carry out standardized treatment for patients according to the severity of the disease, and should adjust medication in time according to the changes of the disease and the treatment response during the treatment process, that is, to implement ladder therapy. In 1995, the National Heart, Lung and Blood Institute (NHLBI) /World Health Organization (WHO) formulated a global strategy for the management and prevention of bronchial asthma, and formed the global initiative for asthma (GINA) [1], which was revised in 2002 [2]. GINA put forward the goals of comprehensive control of asthma, including: minimal (ideally no) asthma symptoms, including nighttime symptoms; Minimal (infrequent) acute attacks (exacerbations) of asthma; No emergency medical treatment; minimal (ideally no) on-demand use of beta 2 receptor agonists; No restricted activity, including exercise; Maximum expiratory flow (PEF) daily variation rate<20%; (near) normal PEF; As early as 1993, China formulated the definition, diagnosis, severity classification and efficacy judgment criteria of bronchial asthma, namely the Chinese guidelines for prevention and treatment of bronchial asthma [3]. Since then, it has been revised twice, and its basic content is consistent with GINA [4].

The treatment of autoimmune diseases has always been the focus and difficulty in this field. Up to now, the etiology of such diseases is not clear, and the pathogenesis is not thoroughly studied. Therefore, the treatment strategies of autoimmune diseases are mainly to alleviate symptoms, such as anti-inflammatory and analgesic, immune regulation and ready-for-use traditional Chinese medicine, etc. However, the inflammation occurring in the pathogenesis of autoimmune diseases has multiple functions, and the simple anti-inflammatory and the use of non-specific immunosuppressants often lead to various serious toxic and side effects. For example, tumor necrosis factor (TNF) and interleukin receptor antagonists that have been used in clinic in recent years often lead to a significant increase in the infection rate of patients, especially tuberculosis infection, and whether this can significantly increase the incidence of malignant tumors remains to be observed in a longer term. It can be seen that, This kind of new generation therapy directly targeting inflammatory cytokines is facing great challenges.Therefore, a safer, more specific and long-term effective therapeutic measure is urgently needed in clinic.With the deepening of gene therapy research in recent 20 years, especially through the discussion of several autoimmune diseases,it is believed that gene therapy as a new therapeutic strategy may bring new prospects for the treatment of autoimmune diseases.At present, many research groups in the world have tried various gene therapy methods in animal models of various autoimmune diseases, such as type 1 diabetes,rheumatoid arthritis (RA) and multiple sclerosis (MS), and there are also reports of clinical phase I ~ II trials. The research results are still exciting.

video-assisted thoracic surgery (VATS), as a minimally invasive surgical technique, has been widely used in all kinds of thoracic surgery and has played an important role [1]. From June 2002 to July 2005, 38 cases of pulmonary diseases were treated with anatomical lobectomy by VATS in the General Hospital of the People's Liberation Army, with satisfactory results.

deep vein thrombosis (DVT) is the main source of pulmonary embolism embolism, which can lead to fatal pulmonary thromboembolism (PTE) and long-term deep vein insufficiency of lower limbs. Elderly patients with lower limb fractures are prone to DVT because of long-term bed rest, old age and immobilization [1, 2]. Therefore, active prevention and treatment of DVT is an important issue to improve the quality of life of elderly patients with fracture and prevent the deterioration of disease.

systemic inflammatory response syndrome (SIRS) is an inflammatory cascade with amplifying effect, and it is an important pathophysiological link and clinical stage of multi-organ dysfunction syndrome (MODS). SIRS is also a common complication after coronary artery bypass grafting (CABG), and it is the pathological basis of complications such as low cardiac output, perfusion lung, respiratory failure, hemorrhagic disease and brain dysfunction after CABG. Accurate assessment of the severity of SIRS after CABG and effective measures to block the progressive development of SIRS can prevent the occurrence and development of MODS, improve the prognosis and improve the survival rate.

Familial febrile convulsion (FC) is a special type of epileptic syndrome in pediatrics, and its pathogenesis is unclear. Studies have confirmed that it has obvious genetic predisposition [1]. In this region, we selected NRTN, CAPS, GPX4 and CSNK1G2 genes as candidate genes according to gene function, and amplified their exons and flanking regions by PCR technique and sequenced them to find mutations.

survivin is an apoptosis inhibitory gene discovered in recent years, which can inhibit cell apoptosis [1]. Human telomerase reverse transcriptase (hTERT) is a restrictive component of telomerase activity. Condyloma acuminatum (CA) proliferates rapidly and often relapses after treatment, and some cases may become malignant, which may be related to the presence of apoptosis and abnormal telomerase activity in CA. However, the expression of survivin in CA has not been reported. In order to study the expression of survivin mRNA and hTERT mRNA in CA tissues, we used reverse transcription-PCR (RT-PCR) and in situ hybridization to detect the expression of survivin and hTERT mRNA in CA tissues, respectively, and explore their possible mechanism in the pathogenesis of CA.

A 52-year-old male was admitted to the hospital on February 18, 2005 due to persistent dull pain in the anterior chest for more than 40 days. Chest X-ray examination after admission showed bone changes in the sternum body (Figure 1); Chest CT showed localized expansion of the joint of the sternal stem body, local cortical destruction, uneven thickness of the cortex bone, soft tissue density shadow in the medullary cavity, and no obvious swelling of the soft tissues of the anterior and posterior walls; However, radionuclide bone scan showed abnormal concentration of radioactivity in the middle sternum.