Volume 87  ·   Issue 06  ·  2007

In recent years, the diagnosis and treatment of coronary heart disease have developed rapidly, and related basic research, drug therapy and interventional therapy have been deepened. However, there are few research reports on cardiomyopathy, and the progress of basic and clinical research is relatively lagging behind other cardiovascular diseases. However, there is no doubt that cardiomyopathy seriously affects the quality of life of patients and endangers their lives, which is one of the main causes of death from cardiovascular diseases.

A large-scale population epidemiological study and a series of large-scale clinical trials of lipid-lowering therapy have confirmed that lipoprotein is independently related to coronary heart disease. However, it is believed that the bottom line concentration of low-density lipoprotein cholesterol (LDL-C) has not been explored yet. A recent meta-analysis suggests that the preventive and therapeutic effect of statin drugs on atherosclerotic cardiovascular disease cannot be attributed solely to its LDL-C reduction effect, but the effects other than lipid-lowering are also very important. Moreover, even if lipid-lowering is intensified, it can only reduce the cardiovascular disease events by 25% ~40%, and there are still 60% ~75% events that cannot be prevented by reducing LDL-C solely. This suggests that we need to find new therapeutic targets, and low-high-density lipoprotein cholesterol (HDL-C), as an important risk factor for coronary heart disease, meets the conditions of being the main therapeutic target. Although early studies have confirmed that HDL-C has a significant negative correlation with the risk of coronary heart disease, and even some studies have found that HDL-C can predict the risk of coronary heart disease better than LDL-C [1], the emphasis on HDL-C is far less than LDL-C, One of the reasons for this is that there are currently no drugs that can effectively and safely raise HDL-C.

Objective to investigate the pathogenic genes of hypertrophic cardiomyopathy (HCM) in China and analyze the relationship between genotype and clinical phenotype. Methods the mutations of cardiac troponin T gene (TNNT2), cardiac myosin binding protein C gene (MYBPC3) and β-myosin heavy chain gene (MYH7) were screened in a HCM family. The exon fragments of the functional regions were amplified by polymerase chain reaction (PCR) and sequenced by dideoxytermination method. The data of the family survey included clinical manifestations, physical examination, cardiac ultrasound and electrocardiogram. Results Among the 10 subjects in this family survey, 4 cases carried TNNT2 14035c>T (R130C) mutation, all of which developed before the age of 40, with a penetrance rate of 100%. There was no abnormality in the same position in the normal control group. The mutation site changed the arginine at position 130 of exon 10 of TNNT2 gene to cysteine. The proband and his two brothers showed mainly cardiac dysfunction. Two brothers of the proband died suddenly during our follow-up.No mutations were found in MYH7 and MYBPC3 genes.Conclusion TNNT2 gene 14035c>t mutation is the pathogenic mutation in this HCM lineage.The clinical phenotype of its carrier is worse.For HCM families with worse clinical phenotype and higher incidence of sudden death, it is necessary to perform TNNT2 mutation screening.

Objective to investigate the activation of transforming growth factor β 1 (TGF β 1) -related Smads signaling pathway and the intervention effect of valsartan in diabetic cardiomyopathy (DCM). Methods Forty male Wistar rats were randomly divided into three groups: control group (8), DCM group (16) and valsartan group (16). DCM rats were fed with high-fat and high-calorie diet plus low-dose streptozotocin (STZ) injection to make DCM rats model. After the model was formed, the valsartan group was treated with valsartan (30 mg/kg). At the end of the experiment, the changes of biochemical indexes and hemodynamic changes of cardiac function of rats were observed. The collagen content, mRNA and protein expression levels of TGF β 1, TGF β ⅡR, Smad2, Smad3 and Smad7 in left ventricular myocardium of rats were detected by electron microscope, Masson staining, real-time quantitative reverse transcription (RT) -PCR and Western blotting techniques. Results The cardiac function of rats in DCM group was significantly lower than that in control group (P<0.01), and the collagen content in left ventricular myocardium was significantly higher than that in control group At the same time, the mRNA expression levels of TGF β 1, TGF β ⅡR, Smad2 and Smad3 were all higher than those in the control group (0.0054 ± 0.0009 vs 0.0126 ± 0.0057, 0.0523 ± 0.0218 vs 0.0877 ± 0.0272, 0.0413 ± 0.0186 vs 0.0884 ± 0.0146, 0.0064 ± 0.0021 vs 0.012 ± 0.0048, P<0.05~0.01), the mRNA ratios of Smad2/Smad7 and Smad3/Smad7 were also significantly higher than those in the control group (P<0.05), the expression levels of TGF β 1, phosphorylated (P) -Smad2 and P-Smad3 proteins were significantly higher than those in the control group (103 ± 18 vs 143 ± 17, 107 ± 21 vs 212 ± 43, 89 ± 17 vs 151 ± 32, P<0.01), and the ratios of P-Smad2/Smad7 and P-Smad3/Smad7 proteins were also significantly higher than those in the control group (P<0.05).

Objective to investigate the relationship between the allele polymorphism of retinoic acid receptor α (RARA) gene D17S579 microsatellite locus and the genetic susceptibility to non-syndromic cleft lip and palate (NSCL/P) in Han population of Hunan, China.Methods PCR and polyacrylamide denaturing gel were used to study the polymorphism of RARA gene D17S579 microsatellite locus in 132 healthy subjects and 140 patients with NSCL/P. Results 10 alleles were detected in the two groups, and the allele frequencies of normal control group and case group were statistically significant (χ 2=17.163, df =9, P =0.046); The allele frequencies of A6 (8.93%) and A9 (13.21%) in the case group were significantly higher than those in the healthy control group A6 (4.17%) and A9 (5.68%, both P<0.05); There was no significant difference in allele frequency between patients with family history and those without family history (χ 2=2.710, P =0.978). Conclusion Microsatellite polymorphism of RARA gene D17S579 is genetically susceptible to NSCL/P in Hunan, China, and alleles A6 and A9 may be risk factors for NSCL/P. There was no association between RARA gene polymorphism and family history of NSCL/P.

Objective to observe the changes of epithelial space widening (DIS) in esophageal mucosa of various subtypes of gastroesophageal reflux disease (GERD) before and after esomeprazole therapy. Methods 19 patients with GERD with heartburn as the main complaint, which were confirmed by endoscopy, 24-h esophageal pH monitoring and proton pump inhibitor therapy, were treated with esomeprazole 40 mg/d for 2 months. Biopsies were taken from normal mucosa at 3~5 cm above the dentate line of esophagus before and after endoscopy, and histology and transmission electron microscopy were performed, respectively. Results After 2 months of treatment, DIS returned to normal in 5 cases (83.33%), 6 cases (85.71%) and 5 cases (83.33%) of erosive esophagitis (6 cases), NERD with pathological acid reflux (7 cases) and NERD without pathological acid reflux (6 cases), respectively, and the symptoms of heartburn disappeared completely. The symptoms of heartburn still existed in 2 of 3 patients with DIS not returned to normal. Conclusion DIS recovered after 2 months of esomeprazole therapy in GERD patients There was no statistically significant difference in DIS recovery between the groups.

Objective to evaluate the efficacy and safety of intragastric water balloon in the treatment of obese patients. Methods 48 obese patients (35 females, mean age (39 ± 9) years) from Hong Kong Special Administrative Region, China, who had received and completed intragastric water balloon treatment since November 2004, were analyzed. The average basal body weight and body mass index of the patients were (106 ± 26) kg and (40 ± 9) kg/m2, respectively, and 34 (70.8%) of them suffered from obesity-related diseases. After intragastric water balloon treatment, the average body weight decreased (13±7) kg; The mean body mass index decreased (5 ± 3) kg/m2, and the mean excess body weight decreased (45 ± 36) %; The mean waist circumference decreased (12 ± 8) cm; All the above indexes were significantly lower than those before treatment (all P<0.01). 32 (66.7%) patients were satisfied with the efficacy of this weight loss method; No serious complications related to intragastric water balloon were found. Conclusion Intragastric water balloon is a safe weight loss device, which can achieve appropriate weight loss effect in Chinese obese patients.

Objective to investigate the psychological status of patients with congenital microtia and to explore the related influencing factors.Methods A cross-sectional survey of 410 patients with congenital microtia was conducted.The patients with psychological disorders were screened by Achenbach Child Behavior Scale (CBCL) and Self-rating Symptom Scale (SCL-90) as standards, and the related factors were analyzed by univariate χ 2 test and multivariate Logistic regression analysis.Results 83 patients (20.2%) had "depression" psychology; 150 patients (36.6%) had "interpersonal sensitivity or social difficulty"; 108 patients (26.3%) had "hostile OR aggressive" psychology. The increase of age (OR =1.75), severity of deformity (OR =2.77), low education level of mother (OR =2.49), ridicule of others (OR =2.58), excessive psychological pressure of family members (OR =1.96), incorrect attitude of family members towards patients (OR =2.45) and long-term family disharmony (OR =2.57) were related to the formation of "depression" psychology of patients; Increasing age (OR =1.62), being ridiculed by others (OR =3.34) and excessive psychological pressure of family members (OR =1.89) were related to the formation of psychology of "sensitive interpersonal relationship OR social difficulty"; Conclusion the risk factors related to the formation of psychological problems in microtia patients should be effectively controlled and avoided, and early education should be given to the patients. It is necessary to carry out external ear reconstruction surgery as soon as possible to prevent psychological problems.

Objective to investigate the role of phosphatidylinositol-3 kinase (PI3K) /Akt signaling pathway in the pathogenesis of ulcerative colitis (UC) and to provide experimental evidence for wortmannin as a new drug for the treatment of UC. Methods 30 patients with UC and 15 normal tissues adjacent to colon cancer were collected as controls, and the expression of phosphorylated Akt (p-Akt) in intestinal mucosa of UC was detected by immunohistochemistry. The effects of wortmannin, an inhibitor of PI3K/Akt signaling pathway, on the expression of tumor necrosis factor (TNF) -α in UC intestinal mucosal biopsy tissues were observed by tissue culture in vitro.Results ①The expression of p-Akt in UC intestinal mucosal tissues was significantly higher than that in the normal control group (A values were 73.6±5.2, 18.0±2.6, respectively, and the positive staining area was 720±58, 133±29, all P<0.05). ②Compared with the UC control group without wortmannin treatment, the level of TNF-α secreted in UC intestinal mucosal tissues after wortmannin treatment was significantly lower (296±39, 138±11, 26±5, all P<0.05). ③The expression of p-Akt in UC intestinal mucosal tissues after wortmannin treatment was lower than that in UC control group without wortmannin treatment (A values were 72.3±6.2, 35.3±5.6, 18.0±2.2, respectively, and the positive staining area was 716±94, 351±50, 129±30, all P<0.05). Conclusion ①PI3K/Akt signaling pathway is involved in pro-inflammatory

Objective to investigate the effect and significance of tetrandrine on pulmonary development and pulmonary hypertension in animal models of congenital diaphragmatic hernia.Methods 25 adult female SD rats were randomly divided into 5 groups, 5 rats in each group.After successful timed breeding, each pregnant rat in the normal control group was given edible oil by gavage on the 9.5th day after breeding, and each rat in the diaphragmatic hernia group was given Nitrofen by gavage on the 18.5th day after breeding, each rat in the normal control group was intraperitoneally injected with normal saline; Each diaphragmatic hernia group was given corresponding intervention measures respectively; All pregnant rats were subjected to cesarean section on the 21st day after breeding, and the lung tissues of fetal rats were removed, and the lung tissues were stained with endothelin immunohistochemistry and image analysis.Results In the normal control group, there were 38 litters of 4 female rats, and there was no diaphragmatic hernia.In the diaphragmatic hernia group, 19 female rats had 126 litters, and 57 had diaphragmatic hernia.The teratogenic rate was 45.2%.The image analysis and statistical data showed that the expression intensity of endothelin in pulmonary arterioles and bronchioles was statistically significant between the diaphragmatic hernia groups.Moreover, there was a positive correlation between the expression intensity of endothelin between pulmonary arterioles and bronchioles, the relative vascular wall thickness, relative vascular wall area and the relative integral optical density of pulmonary arterioles (r =0.847, 0.851, 0.844, all P<0.01). Conclusion Tetrandrine can significantly improve the lung tissue dysplasia, reduce pulmonary hypertension and reduce the expression intensity of endothelin in the lung in the rat model of congenital diaphragmatic hernia.Tetrandrine may inhibit the synthesis and release of endothelin in lung tissue, Inhibiting the binding of endothelin to the corresponding receptor to exert its effect.

Objective to investigate the effect of survivin antisense oligonucleotide (survivin-ASODN) on the apoptosis of bladder cancer cell line BIU87 induced by docetaxel (Taxotere). Methods the successful eukaryotic expression vector pcDNA3-SVVas of survivin-ASODN was transfected into bladder cancer cell line BIU87 by liposome mediation, and the successful transfected positive clones were screened. The expression of survivin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The effect of survivin-ASODN combined with taxotere on the survival of BIU87 cells was observed by trypanosome blue rejection method; The sensitivity of transfected cells to Taxotere was determined by cell counting and tetramethylazo salt (MTT) assay; Analysis of apoptotic DNA fragmentation by agarose gel electrophoresis; Nuclear staining was used to detect the changes of apoptotic cells' nuclei; Results the expression level of survivin mRNA in BIU87-SVVas transfected with pcDNA3-SVVas was decreased, and cell proliferation was significantly inhibited, compared with that in BIU87-neo transfected with pcDNA3 empty vector and BIU87 without vector, the differences were statistically significant (P<0.05). By agarose gel electrophoresis, DNA trapezoidal bands were observed in BIU87-SVVas cells, but not in other controls; Compared with BIU87-neo and BIU87 cells, the nuclei of BIU87-SVVas cells were densely stained; The apoptosis rate of BIU87-SVVas cells treated with Taxotere increased greatly. Conclusion Survivin-ASODN can promote the apoptosis of BIU87 cells induced by Taxotere and increase its sensitivity to Taxotere, which laid the experimental foundation for the biological therapy of bladder cancer.

Objective to investigate the feasibility of inducing the differentiation of human bone marrow-derived pluripotent adult progenitor cells (ZHJ-MAPC) into hepatocytes under the condition of co-culture with human hepatocyte line L02 in vitro.Methods (1) Indirect co-culture: ZHJ-MAPC and human hepatocyte line L02 were indirectly co-cultured only in the liquid phase.The expressions of albumin, alpha-fetoprotein (AFP), cytokeratin-18 (CK-18) and cytokeratin-19 (CK-19) of ZHJ-MAPC were identified by immunocytochemistry on the 1st, 3rd, 5th and 7th days of culture respectively.2) Direct co-culture: ZHJ-MAPC and L02 cells (both 1×104/ml) labeled with fluorescent dye carboxyfluorescein diacetate succinimide ester (CFSE, green) were mixed and directly co-cultured at a ratio of 50% each. After 5 days, the expression of albumin, AFP, Albumin peaked on day 5; CK-18 was positive on the fifth day, and strongly expressed on the seventh day. CK-19 was negative at all time points. (2) Direct co-culture results: On the fifth day of co-culture, the positive cells with yellow fluorescence, that is, ZHJ-MAPC differentiated into hepatocytes, but the positive expression of AFP was less. Conclusion ZHJ-MAPC derived from human bone marrow can be induced to differentiate into mature liver-like cells by indirect or direct co-culture with human hepatocyte line L02.

Objective to observe the effects of lipiodol hydroxyapatite nanoparticles (nHAP) infusion into the hepatic artery on the growth, necrosis rate, apoptosis index, microvessel density (MVD) and proliferating cell nuclear antigen (PCNA) expression of VX2 liver tumors in rabbits, and to observe their effects on the survival time of rabbits with tumors. Methods Eighty New Zealand white rabbits were randomly divided into four groups, 20 in each group, 2 weeks after implantation of intrahepatic tumors. The rabbits were administered by perfusion through the hepatic artery. The experimental groups were normal saline group (Group A), lipiodol group (Group B), doxorubicin lipiodol group (Group C) and lipiodol nHAP group (Group D). At 1 and 2 weeks after treatment, the tumor size was measured by CT, and the tumor growth rate was calculated. Two weeks after treatment, the necrosis rate of tumor area was observed by pathology, the apoptosis index of tumor was measured by DNA gap end labeling (TUNEL), and the MVD value of tumor was measured by immunohistochemistry. The survival time of rabbits after treatment was recorded in each group. Results At 1 and 2 weeks after treatment, The tumor volume and growth rate in the lipiodol nHAP group were significantly smaller than those in other groups (all P<0.05). Two weeks after treatment, the tumor necrosis rate and apoptosis index in the lipiodol nHAP group were higher than those in other groups (all P<0.05). After embolization, the MVD of residual tumor area in the lipiodol group and the doxorubicin lipiodol group increased by 34±7 and 35±8, respectively, which were higher than those in the negative control group A (23±6, P<0.05); The MVD (16 ± 4) of residual tumor area in lipiodol nHAP group was lower than that in other groups, the proliferation index of embolization group was lower than that in negative control group A, and the proliferation index of lipiodol nHAP group was lower than that in other groups (all P<0.05). The survival days (d) of tumor rabbits in the four groups were 38 ± 6, 46 ± 8, 50 ± 8, 55 ± 9, respectively. The survival time of lipiodol nHAP group was longer than that in other groups (all P<0.05). Conclusion Lipiodol nHAP infusion into hepatic artery can significantly inhibit tumor growth and prolong the survival time of tumor rabbits, which may exert anti-tumor effects by promoting tumor cell necrosis, inducing tumor cell apoptosis, inhibiting tumor cell proliferation and inhibiting tumor angiogenesis.

Objective to investigate the effect of nucleosome binding protein 1 (NSBP1) gene on the proliferation of human hormone-dependent prostate cancer cell line LNCaP by RNA interference (RNAi) technique. Methods Four small molecule interfering RNAs (siRNAs) targeting NSBP1 (including one negative control) were designed and synthesized, and the recombinant pSilencer 2.1-U6 neo plasmid, which can inhibit the expression of NSBP1 mRNA, was constructed and transfected into LNCaP cells. The inhibition efficiency of different plasmids on NSBP1 expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The plasmid with the highest inhibition efficiency was selected and transfected into LNCaP cells. The cell proliferation activity was measured by tetramethylazolazole salt (MTT) assay, and the cell cycle changes were detected by flow cytometry. Results The plasmid pSilencer-81 with the highest inhibition efficiency (80% inhibition of mRNA level and 85% inhibition of protein level) was screened and transfected with the negative control pSilencer-Neg plasmid respectively. The percentage of G2M + S phase cells in LNCaP/81 cells was significantly lower than that in LNCaP/Neg cells (t =3.705, 3.887, 8.220, all P<0.05). Conclusion the siRNA targeting NSBP1 can significantly inhibit the proliferation of prostate cancer LNCaP cells by inhibiting the expression of NSBP1 gene in prostate cancer LNCaP cells.

Methods Four plasmids including pmRNA fluorescein (Luc), pmRNA IRES-Luc, pmRNA chicken ovalbumin (OVA) and pmRNA IRES-OVA were constructed, and the protein expression levels of different forms of Luc mRNA transfected into mouse dendritic cells (DC) were detected by Luc reporter system. The antigen-specific cellular immune responses induced by Capped-OVA mRNA and IRES-OVA mRNA modified DCs were compared by cytotoxic T lymphocyte (CTL) killing experiment and tumor bearing experiment in vivo using OVA as target antigen and mice melanoma lung metastasis as animal model. Results the IRES sequence was inserted into the cDNA sequence of the mRNA transcription template, but the production of the corresponding mRNA was not affected in vitro. IRES-Luc mRNA and Capped-Luc mRNA could still detect Luc activity after transfection of DCs for 96 h. After immunization with Capped-OVA mRNA and IRES-OVA mRNA modified DCs for 1 week, CTL killing rate of 4 h CTL was (28±3) % in Capped-OVA mRNA/DC immunization group, while that of IRES-OVA mRNA/DC immunization group was (32±4) % in 4 h CTL killing rate.

A 29-year-old woman suffered from paroxysmal headache, palpitation and hyperhidrosis for 1 year and aggravated for 2 months. A year ago, the patient developed headache, palpitation and hyperhidrosis after activity. Each attack lasted for 20 min and relieved spontaneously, 4 times/week. During this period, blood pressure was found to be 160/100 mm Hg (1 mm Hg =0.133 kPa), which was not paid attention to. Two months ago, the attacks were frequent, once a day, and the maximum blood pressure measured was 210/160 mm Hg, accompanied by chest, abdomen and neck pressure and near-death sensation, which lasted for 20 min and relieved spontaneously, and blood pressure dropped to 120/80 mm Hg. B-ultrasound showed that the left adrenal gland was enlarged with unclear boundary. There were no symptoms such as increased nocturia, weakness of both lower limbs, soft paralysis, convulsions, etc. No history of kidney disease and no family history of hypertension. Physical examination: body temperature 36.6℃, heart rate 80 beats/min, breathing 20 beats/min, blood pressure 170/140 mmHg of right upper limb, 160/90 mm Hg of left upper limb and 150/100 mm Hg of both lower limbs.

tissue engineering is a new discipline that combines cell biology and material science to construct tissues or organs in vivo and in vitro. The basic method is to combine a certain number of seed cells with specific functions and degradable porous biological scaffold materials and implant them in vivo after a period of in vitro culture. The scaffold materials are gradually degraded and absorbed in vivo, while the seed cells divide and proliferate continuously, synthesize and secrete extracellular matrix, and finally the scaffold is completely degraded to form new tissues or organs with normal structure and function.

Subarachnoid hemorrhage (SAH) is a common type of stroke, which is more common in relatively young and middle-aged people, and the prognosis is generally poor.Delayed ischemic neurological deficit (DIND) often appears 4~10 days after hemorrhage, which is one of the main causes of death and disability in SAH patients.Although nimodipine is used clinically to prevent vasospasm after SAH to alleviate DIND, the efficacy is generally limited. Recently, some authors have found that DIND has an obvious relationship with microembolic load through pathological anatomical data. It has been reported in literature that platelet activation is related to vasospasm and DIND induced by SAH, and platelet aggregation, thromboxane B2 release and thromboxane A2 metabolism increase in SAH patients on the third day.

Cardiomyopathy is a group of diseases manifested by cardiomyopathy. The public and medical workers' understanding of this disease has been troubled by its definition and classification for a long time. The classification of this disease (many in the past) is very important for further understanding this disease; Moreover, it is necessary to use precise language to describe the disease.