Volume 86  ·   Issue 44  ·  2006

The antiviral treatment of AIDS has experienced many stages of development. The earliest antiviral drug used for the treatment of human immunodeficiency virus (HIV) infection is the nucleoside reverse transcriptase inhibitor zidovudine (AZT), which has a certain inhibitory effect on HIV replication. However, many infected patients will develop different degrees of drug resistance after 12 weeks of treatment, and the viral load will rebound. After entering the 1990s, non-nucleoside antireverse transcriptase inhibitors and protease inhibitors have been developed, and the kinds of drugs used in clinic have also increased, and the combination of two drugs has begun to apply. The antiviral effect is strengthened, and the proportion of drug resistance has also decreased, but the long-term curative effect can still not be maintained. In 1996, He Dayi, a Chinese-American, proposed highly active antiretroviral therapy (HAART), that is, the combination of three drugs, which is another milestone in the antiviral treatment of AIDS. The advantage of HAART therapy is that the antiviral activity is further improved, and the viral load in the infected patients is reduced below the detectable level. Obviously delaying the development of drug resistance is still the main means of anti-HIV.

Objective to observe the instability of atherosclerotic plaque in rabbit aorta and the intervention effect of Tongxinluo and simvastatin on it, and to explore its molecular mechanism.Methods Twenty-three male white rabbits with large ears were fed with aortic intima balloon injury and high cholesterol for 16 weeks, and randomly divided into model group and Tongxinluo (1 g·kg-1·d-1) and simvastatin (2 mg·kg-1·d-1) intervention group; The changes of serum lipid, serum endothelin (ET) and nitric oxide (NO) levels before and after the intervention were measured in the normal group (NO drug administration, normal diet feeding for 8 weeks). The expressions of metalloproteinase-1 (MMP-1) and cyclooxygenase-2 (COX-2) proteins and apoptosis-related genes (FasL, Bcl-2) mRNA in the model group were measured by pathological analysis. Results Compared with the normal group, the aortic intima thickness, macrophage content in plaque, MMP-1 and COX-2 expressions in the model group were significantly increased (P<0.05), while the collagen content was significantly decreased (P<0.05). Compared with the model group, the vascular endothelial function of Tongxinluo and simvastatin intervention group was improved, and the aortic intima thickness (0.25 ± 0.13, 0.28 ± 0.12 vs 0.60 ± 0.37), macrophage content in plaque, MMP-1 (4.08 ± 1.11, 5.28 ± 2.36 vs 9.67 ± 0.48) and COX-2 (6.97 ± 1.57, 3.62 ± 0.66, 3.78 ± 1.29) expressions were significantly decreased (P<0.05), while the collagen content was significantly increased (0.11 ± 0.08, 0.22 ± 0.11 vs 0.01 ± 0.01, P<0.05). The expression of FasL mRNA in aortic tissues of the two intervention groups was significantly decreased (0.47±0.36, 1.32±0.61 vs 2.44±0.44, all P<0.05), and the expression of Bcl-2 mRNA was significantly increased (0.64±0.16, 1.66±0.94 vs 0.17±0.11, all P<0.05). There was no significant difference between the two groups (all P>0.05). Conclusion Tongxinluo and simvastatin have the same effect on promoting the stabilization of atherosclerotic plaque, and the curative effect is comparable. The mechanism may be related to inhibiting the activity of COX-2 enzyme, reducing the expression of MMP-1 and apoptosis in plaque.

Objective to investigate the clinical significance of the expression of P-glycoprotein (P-gp), lung drug resistance protein (LRP) and multidrug resistance-associated protein (MRP) in invasive breast cancer tissues. Methods Flow cytometry (FCM) was used to quantitatively analyze the expression of the above proteins in cancer tissues and corresponding adjacent tissues of 68 patients with primary invasive breast cancer. Results The relative fluorescence intensity (RFI) of P-gp, LRP and MRP expression in invasive breast cancer tissues was 0.58, 0.51 and 0.56, respectively, while the corresponding adjacent tissues were 0.26, 0.33 and 0.30, respectively. The expression of drug resistance-associated protein in cancer tissues was significantly different from the corresponding adjacent tissues (P<0.05). There was no correlation between the expression of the three drug resistance-associated proteins and the age, tumor size, pathological type, clinical stage, estrogen and progesterone receptors of patients (P>0.05). The expression of P-gp and MRP was 0.61 and 0.69 in patients with lymph node metastasis, respectively, and 0.54 and 0.45 in patients without lymph node metastasis, respectively There is a positive correlation between the expression of LRP and MRP in breast cancer tissues (P<0.05). Conclusion There are differences in the clinical significance of the expression of three drug resistance-related proteins in invasive breast cancer tissues. The expression of LRP is related to lymph node metastasis and positively correlated with MRP expression, which can be used as a prognostic index of breast cancer.

Objective to explore the principle of scraping and suction anatomy, and to explore and summarize the application skills of scraping and suction anatomy. Methods 2213 cases of complex abdominal surgery performed by scraping and suction anatomy from November 1997 to May 2006 were retrospectively analyzed, and the success rate, operation duration, intraoperative bleeding volume and postoperative complications were statistically analyzed. The average bleeding volume during pancreaticoduodenectomy was 105 ml, and the average operation time was 3.6 h; The rate of cholecystectomy in acute cholecystitis was 100%, the average bleeding volume during radical gastrectomy was 75ml, and the average operation time was 2.3 h. None of them needed postoperative blood transfusion. None of them had major bleeding and collateral injury in retroperitoneal tumor surgery. Conclusion Peng's multifunctional surgical dissection device (PMOD) and the extended scraping and aspiration separation dissection method have shown the superiority in abdominal surgery.

Objective to investigate the role of mitogen-activated protein kinase (MAPK) signal transduction pathway in the synthesis of transforming growth factor β 1 (TGF-β 1) in renal tubular epithelial cell line (HKC) promoted by aldosterone (ALDO). Methods HKC cells were cultured in vitro. (1) The expression of phosphorylated ERK1/2 and total ERK1/2 in cell lysates was detected by Western blot. The relative expression of ERK1/2 (phosphorylated ERK1/2/total ERK1/2) was correlated with the synthesis of TGF-β 1 under the corresponding stimulation conditions. (3) After different concentrations of spironolactone and glucocorticoid receptor blocker RU486 were treated with HKC cells, the expression of phosphorylated ERK1/2 and total ERK1/2 was stimulated by exogenous ALDO However, the synthesis of TGF-β 1 in HKC under 10-7mol/L ALDO stimulation was (121±10) pg/ml, and the difference was statistically significant (P<0.05). However, the synthesis of TGF-β 1 in JNK and P38 branch pathway blockers (SP600125 and SB203580) did not change significantly (all P>0.05). (2) Exogenous ALDO could increase the relative expression of ERK1/2 in HKC in a dose-dependent manner. The relative expression of ERK1/2 in 10-9 and 10-7mol/L ALDO stimulation groups were 0.67±0.06 and 0.80±0.05, respectively, which were significantly higher than those in 0 mol/L ALDO group (P<0.05 or 0.01). There was a significant positive correlation between the relative expression of ERK1/2 and the synthesis of TGF-β 1 under the corresponding concentration of aldosterone stimulation (R =0.793, P<0.01); After 10-7mol/L ALDO stimulated HKC for 15 min, phosphorylated ERK1/2 began to increase significantly and reached a peak (P<0.01), and its relative expression level was 0.84±0.06, then gradually decreased. At 360min, its expression level was 0.49±0.08, which was not statistically significant compared with that of 0min (P>0.05). (3) When 10-7mol/L ALDO was incubated with 10-9 and 10-7mol/L spironolactone for 30 min, the relative expression level of ERK1/2 was 0.62±0.08 and 0.60±0.04, respectively, which was significantly lower than that of 0 mol/L spironolactone group (P<0.05 or P<0.01), but its expression level was still significantly higher than that of the control group without any stimulation (P<0.05); The expression of ERK1/2 was not significantly lower than that in the 0 mol/L RU486 group when co-stimulated with 10-7mol/L ALDO and RU486 for 30 min (P>0.05). Conclusion ERK1/2 signal transduction pathway may be one of the main pathways that mediate the synthesis of TGF-β 1 in HKC promoted by aldosterone, and the binding of aldosterone to intracytoplasmic receptors is an important condition for its pathological role by activating ERK1/2 pathway.

Objective to observe the effects of insulin on cardiac function, coronary blood flow, coronary diastolic function and apoptosis of coronary endothelial cells in dogs with acute myocardial ischemia/reperfusion (MI/R). Methods A canine MI/R model was prepared, with quantitative myocardial ischemia (left anterior descending artery blood flow decreased by 80%) for 50 min and reperfusion for 4 h, and was randomly divided into normal saline, glucose-insulin-potassium solution (GIK), glucose-potassium solution (GK) and sham operation groups. After reperfusion, the coronary blood vessels were separated, and the coronary diastolic function was observed by isolated vascular perfusion. At the same time, the content of NO in the perfusion was measured, and the apoptosis of coronary endothelial cells was detected by in situ end labeling method. Results Compared with the saline group, the left anterior descending coronary blood flow (CBFLAD) was increased in the GIK group (19.2 ml/min±2.2 ml/min vs 14.6 ml/min±1.8 ml/min, P<0.05), and the left ventricular systolic and diastolic function was improved after reperfusion, while the GK group had NO such effect. Compared with the sham operation group, the MI/R saline group had obvious coronary diastolic dysfunction, decreased NO release and massive apoptosis of coronary endothelial cells. The GIK group could effectively protect the coronary diastolic function (P<0.01) and increase the release of NO (P<0.01). At the same time, it was found that insulin significantly inhibited the apoptosis of coronary endothelial cells induced by MI/R (12%±4%), and the normal saline group (45%±7%, P<0.01). Conclusion Insulin administration during reperfusion can significantly reduce the coronary artery damage induced by MI/R Inhibition of apoptosis of MI/R coronary vascular endothelial cells.

Objective to study the immune effect of heparinase gene-modified dendritic cell (DC) vaccine on gastric cancer cells and to explore the feasibility of heparinase vaccine in active immunotherapy of gastric carcinoma. Methods DCs derived from peripheral blood mononuclear cells of human leukocyte antigen A2 (HLA-A2) positive healthy volunteers were infected with recombinant heparinase adenovirus containing full-length heparinase cDNA. The DC vaccine modified by heparinase gene was prepared. The lymphocytes from the same source were stimulated to activate them into heparinase-specific cytotoxic T lymphocytes (CTL). The immune effect of heparinase-specific CTL on KATO-Ⅲ and SGC-7901 gastric cancer cells in vitro was verified by standard 51Cr release assay. The release of interferon (IFN) -γ after heparinase-specific CTL was co-incubated with different target cells was performed by enzyme-linked immunosorbent assay (ELISA). Results the expression of heparinase protein in the DC was significantly increased after heparinase recombinant adenovirus infection. On the contrary, SGC-7901 had no killing effect on heparinase-positive but HLA-A2-negative gastric cancer cells, and its killing activity was only 11.1% ± 4.6% even at the highest efficiency/target ratio; Further studies showed that heparinase-specific CTL had no immune killing effect on autologous lymphocytes, and its killing activity was 11.4% ± 7.9% at the highest efficiency/target ratio. At the same time, it was also found that the expression of IFN-γ in heparinase-modified DC-stimulated effector cells was significantly higher than that in empty virus-modified group and IL-2-stimulated group after co-incubation with KATO-Ⅲ (280 pg/ml ± 24 pg/ml vs 121 pg/ml ± 19 pg/ml and 60 pg/ml ± 11 pg/ml, P<0.05); However, there was no significant difference in IFN-γ detected by specific effector cells stimulated by heparinase modified DC and SGC-7901 or autologous lymphocytes (all P>0.05). Conclusion Heparinase modified DC vaccine can induce specific CTL and has good immune killing activity against HLA-matched and heparinase-positive gastric cancer cells, but has no immune killing effect on autologous lymphocytes, so it is a safe and effective target for tumor immune gene therapy.

Objective to investigate the relationship between serum anti-β 2 glycoprotein Ⅰ antibody (A β 2gpⅠ) and platelet activation status in patients with ulcerative colitis (UC). Methods A case-control study was performed, using enzyme-linked immunosorbent assay (ELISA) and flow cytometry technique to detect the serum A β 2gpⅠ level and the positive rate of platelet activation markers in 56 patients with UC and 25 healthy subjects, respectively, and the relationship between the two levels and the severity of active UC was analyzed. Results ①The level of IgG type A β 2gpⅠ was statistically significant compared with the active UC group (0.61±0.13), the remission UC group (0.50±0.13) and the normal control group (0.22±0.14); The positive expression rate of fibrinogen receptor (PAC) -Ⅰ and P-selectin (CD62P) on the platelet surface of active UC patients (30.6%±7.6%, 45.0%±8.8%) was significantly higher than that of normal control group (6.3%±1.7%, 9.2%±2.7%) and remission UC group (19.6%±7.8%, 31.9%±7.8%), and the differences were statistically significant; The difference was statistically significant. ③ Using one-way analysis of variance, the level of A β 2gp Ⅰ (IgG), the positive expression rate of PAC-Ⅰ and CD62P [FA β 2gpi =3.679, FPAC-I (%) =5.346, FCD62P (%) =5.418] were significantly different in the active UC patients for different disease severity. Conclusion A β 2gp Ⅰ and platelet activation status were closely related to the occurrence and development of UC.

Objective to investigate the probability and mutation pattern of connexin 26 gene (GJB2) in patients undergoing cochlear implantation. Methods 115 Chinese patients undergoing cochlear implant and 109 healthy controls were collected. Genomic DNA was extracted from peripheral blood and analyzed by polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) and direct sequencing. Results GJB2 gene mutation was found in 36.5% (42/115) of cochlear implant patients, 41% (41/100) of non-syndromic deafness patients, and only 1 case (1/15) of inner ear malformation group. There were 11 mutation patterns found in the study, among which 235delC was the most common mutation type, and the allele frequency accounted for 18.3% (42/230) of all cochlear implant patients and 21.0% (42/200) of non-syndromic deafness group. 187G>T and 230G>A mutations were the first new mutations found. Conclusion GJB2 gene mutation is the main deafness-causing gene mutation in Chinese cochlear implant patients

Objective to explore the strategy of effective activation of mucosal and systemic immunity in BALB/c (H-2d) mice immunized with human immunodeficiency virus type 1 (HIV-1) vaccine.Methods BALB/c mice aged 6-8 weeks were randomly divided into 6 groups, 3 mice in each group; DNA vaccine with or without vaccine adjuvant was used for mucosal-guided immunization, and recombinant vaccinia Tiantan strain vaccine was used for systematic boosting strategy to improve the body's immune response. The immunization time of DNA vaccine was 0, 14 and 28 days, and the dose was 10 μ g/time per mouse; The immune response of T lymphocytes was measured by ELISPOT and intracellular staining on the 56th day after the first immunization. Results Only weak systemic T cell immunity (2 ± 1) spot forming cells (SFC/106 cells) were activated after mucosal immunization with DNA vaccine alone. Co-administration of cholera toxin to mucosa could improve the corresponding systemic T cell immunity (14 ± 11 SFC/106 cells). After the recombinant vaccinia Tiantan strain vaccine system boosted, the specific T lymphocytes secreting γ-interferon (IFN-γ) increased 4.5 times (61 ± 35 SFC/106 cells). The immune response of HIV-1 gag-specific CD8+ T cells could be obtained by intracellular staining, and the levels of specific IgG antibody in serum and specific IgA antibody in pulmonary lavage fluid increased by 2 times [absorbance (A) value: 1.5±0.3] and 2.5 times (A value: 1.8±0.8) respectively after systemic boosting with vaccinia Tiantan strain. Conclusion Mucosal guided immunization with systemic boosting can induce effective mucosal, systemic humoral immune response and T lymphocyte immunization.

Objective to study the variation characteristics of the full-length gp120 gene of recombinant human immunodeficiency virus type 1 (HIV-1) AE (CRF01_AE) strain, and to provide help for the immunological study and vaccine development of recombinant CRF01_AE. Methods Nested PCR was used to amplify the full-length gp120 of outer membrane protein from 21 blood samples from 100 HIV-1 CRF01_AE infected patients in Fujian Province from 2004 to 2005. The sequence of the full-length gp120 of HIV-1 was analyzed by MEGA, DNASIS, CLUSTAL1.83 and N-GLYCOSITE. Results The gene phylogenetic tree showed that the 21 samples belonged to CRF01_AE recombinant HIV-1 strain, and the gene distance in the sample group was 9.5% ± 2.5%. There were four types of tetrapeptides at the apical end of V3 loop: GPGQ (71.43%), GPGR (19.05%), GPGH (4.76%) and GQGQ (4.76%) Conclusion Most of the recombinant strains of CRF01_AE in Fujian Province are closely related to the representative strains of Southeast Asia, and their origins are complex. Most of the recombinant strains of CRF01_AE are NSI type, and the gp120 gene variation is large. This study provides rich materials and basic data for the future research on virus immunology and vaccine.

Objective to investigate the characteristics of HIV-1 gag-specific T lymphocyte proliferation response in human immunodeficiency virus (HIV) infected subjects, analyze their relationship with disease progression, and explore the mechanism of HIV-specific immunodeficiency. Methods The frequency of specific proliferation response of PBMC to HIV-1 Gag antigen in 34 HIV-1 infected subjects was detected by BrdU-doped flow cytometry intracellular staining method, and their correlation with absolute value of CD4+ T cells, plasma viral load and CD8+ T cell activation index was analyzed. Results The proliferation rate of HIV-1 gag-specific CD4+ T cells in HIV infected subjects was significantly lower than that in healthy controls (P<0.01), and it was negatively correlated with CD38 expression of CD8+ T cells (P<0.01), positively correlated with absolute value of CD4 (P<0.01), and negatively correlated with plasma viral load (P<0.05); The proliferation rate of HIV-1 gag-specific CD8+ T cells in Chinese HIV infected patients was significantly higher than that in healthy controls (P<0.01), and it was positively correlated with CD38 expression of CD8+ T cells (P<0.01), significantly negatively correlated with absolute value of CD4 (P<0.01), but not correlated with plasma viral load (P>0.05). Conclusion The proliferation function of HIV-1 gag-specific CD4+ T cells in HIV infected patients was abnormal, and the proliferation function of HIV-1 gag-specific CD4+ T cells was decreased, and its degree was closely related to the disease progression.

Humans mainly rely on natural immunity and acquired immune response to recognize and defend against microbial infection or distinguish "non-self" antigens. Among them, acquired immune response is delayed and antigen-specific, mediated by antigen receptors on B and T lymphocytes, and has immune memory function. Acquired immunity starts when T cells recognize exogenous peptides restricted by histocompatibility antigen (MHC) expressed in antigen presenting cells (APC), but T cell activation requires not only the recognition of MHC complex of exogenous peptides, but also the expression of co-stimulatory molecules such as CD80 and CD86 on APC. In order to ensure the normal activation of T cells, the recognition of T cells must be strictly regulated, among which the strict expression of co-stimulatory molecules is the key to regulation, but this regulatory mechanism is still not clear enough. In 1989, Janeway [1] speculated that the regulation of co-stimulatory molecules must be controlled by the characteristic receptors that recognize microbial products, The recently discovered Toll-like receptors (TLR) family of autoimmune key receptors is in line with this hypothesis, and a series of studies have shown that they play an important role in controlling the acquired immune response.

Since the first case of AIDS was reported in the United States in 1981, AIDS has become the most serious and catastrophic disease facing mankind. According to<At present, there are 38 million HIV carriers in the world, among which the number of HIV infections in Asia is the fastest increasing. In 2003, a total of 2.9 million people died of AIDS [1]. Since 1995, human immunodeficiency virus 2 (HIV) infection has entered a widespread epidemic period in China, and it has been at the critical point of spreading from high-risk population to general population [2]. With the widespread application of antiretroviral therapy, the survival time and opportunistic infection of HIV infected patients have been controlled to a large extent, while non-infectious complications including pulmonary hypertension have received more and more attention in the world, and related studies are increasing. If patients with pulmonary hypertension are complicated, the survival time will be significantly reduced, in fact, pulmonary hypertension has become an independent risk factor for death of HIV infected patients [3]. Since the late 1980s, there have been a lot of studies on epidemiological investigation, pathogenesis and treatment methods of HIV infection complicated with pulmonary hypertension abroad, However, only one case of HIV-associated pulmonary hypertension (PAHRH) has been reported in Fuwai Cardiovascular Hospital [4], and the relevant research in this area is almost blank. This article will elaborate the epidemiological data, pathogenesis, clinical characteristics and treatment of PAHRH.

A 32-year-old female was admitted to the hospital due to repeated pain in the right waist and right lower abdomen for 1 year, which aggravated for 1 month. The patient developed distension and pain in the right waist and right lower abdomen without obvious inducement 1 year ago, and the B-ultrasound in the local hospital showed right kidney stones. A month ago, the patient's symptoms aggravated, persistent pain, accompanied by macroscopic hematuria, no nausea, vomiting, no frequent urination, urgency and painful urination. Physical examination: mild buckling pain in the right kidney, but no obvious positive signs were found. Urinary routine showed red blood cells (+ + + + + +), and other laboratory examinations (-). CT findings: plain scan showed a round-like isodensity soft tissue mass in the anterior part of the upper pole of the right kidney. The size of the mass was 46mm ×41mm ×55mm. The edge of the mass was clear, but no capsule was seen. The mass protruded forward outside the renal contour and pressed the renal pelvis and calyces posteriorly, causing moderate hydrops in the renal pelvis and calyces on the affected side (Figure 1A). Enhanced scan: the mass showed mild uneven enhancement, and multiple enlarged lymph nodes were seen on the right side of the abdominal aorta, which fused into a mass, with local low density foci, which pushed the inferior vena cava forward (Figure 1B).

The pedicled transverse island foreskin flap (Duckett operation) and scrotal septal pedicled flap are commonly used in clinical practice because of their high success rate. From May 1994 to October 2005, we used these two methods and designed a simple and practical one-stage orthopedic repair of 76 cases of hypospadias in children with double perineal drainage. Different pedicled tissue flaps were used for different types of hypospadias, and the curative effect was remarkable.

Due to the structure and pathophysiological characteristics of the ear, the incidence of acute otitis media in children is much higher than that in adults, and it is one of the common diseases causing fever in children. Acute otitis media in children often lacks early specific clinical manifestations. Therefore, in European and American countries, pediatricians routinely perform tympanic membrane examination for children with fever to determine whether there is middle ear infection. However, in China, there is no clear regulation of pediatric diagnosis and treatment routine. Is the incidence of acute otitis media in Chinese children indeed lower than that in European and American children? Should children with fever have routine otoscopy? Therefore, we conducted an epidemiological investigation of acute otitis media in children with fever in our hospital from March to August 2004. The results are reported as follows.

Helicobacter pylori (Hp) infection is closely related to the occurrence of gastritis, peptic ulcer, gastric cancer and other diseases in human body [1,2]. Immunization is the most effective measure to control the epidemic of Hp infection [3]. However, due to the complex immune mechanism of Hp infection, no vaccine antigen with ideal effect has been found yet [4]. The outer membrane of Hp, as an important target of immune attack, plays an important role in human protective immune response [4,5]. Omp11 is an outer membrane protein of Hp, and its encoding gene is highly conserved, which is expected to become a vaccine antigen [6]. However, no reports on the activity and immune effect of Omp11 antigen have been retrieved at home and abroad. Therefore, it is proposed to use gene cloning method to study the preparation, antigenic activity and application value of Omp11 recombinant protein (rOmp11).

A 4-year-old male was admitted to the hospital on November 28, 2005 due to the diagnosis of hepatomegaly half a month ago. During routine physical examination half a month ago, the child was found to have hepatomegaly, negative hepatitis B and C virus markers, alanine aminotransferase (ALT) 140 U/L and glutamate aminotransferase (AST) 180 U/L. Physical examination after admission: body temperature 36.5℃, pulse 82 beats/min, breathing 18 beats/min, blood pressure 90/56 mm Hg (1 mm Hg =0.133 kPa), body weight 15kg. He was conscious and had no special disease. No yellowing and bleeding spots on the skin, no enlargement of superficial lymph nodes, clear breathing sounds in both lungs, normal heart boundary, full abdomen, no distension in abdominal wall veins, the upper boundary of liver was located in the 6th intercostal line of the middle clavicle, and the lower boundary of liver was located about 40 mm below the costal line of the middle clavicle, with mild tenderness and no palpation of the spleen. Neurological examination: cervical resistance (-), Buchner sign (-), Klinker's sign (-), cranial nerve examination showed no positive signs, muscle tone of limbs was normal, muscle strength was grade IV, physiological reflex was symmetrical, but pathological reflex was not drawn out.