Volume 86  ·   Issue 20  ·  2006

In recent years, the incidence of infertility has been increasing gradually. With the widespread application and continuous improvement of assisted reproductive technology (ART), the clinical treatment of infertility has made great breakthroughs, but this has also hindered the research in the basic fields of reproductive physiology, reproductive endocrine and infertility to some extent [1]. Some research methods are too simple or lack of uniform standards, which makes many important basic research results cannot draw appropriate conclusions or give scientific and reasonable explanations.

Objective to observe the effect of pulmonary artery perfusion with low temperature washing erythrocyte protective solution on lung injury during open heart surgery. Methods Thirty patients with rheumatic mitral valve disease accompanied by moderate to severe pulmonary hypertension (PH) were selected and divided into pulmonary artery perfusion group and control group (15 cases each). After ascending aorta occlusion, the perfusion group was intermittently perfused with low temperature washing erythrocyte protective solution through the aortopulmonary artery. The perioperative pulmonary vascular resistance (PVR), the ratio of venous blood to arterial blood leukocyte count (V/A), the content of malondialdehyde (MDA) and oxygenation index (OI) in pulmonary circulation, and the time of mechanical assisted ventilation were measured and recorded in the two groups. Results (1) PVR: Immediately after bypass, 12 and 24 hours after operation, the perfusion group (46.4 kPa·s·L-1±8.1 kPa·s·L-1,48.5 kPa·s·L-1±7.0 kPa·s·L-1,36.1 kPa·s·L-1±6.3 kPa·s·L-1) was significantly lower than that in the control group (65.7 kPa·s·L-1±5.3 . (2) MDA contents were significantly lower in perfusion group (14.3 mmol/L ± 0.8 mmol/L, 16.1 mmol/L ± 0.7 mmol/L, 13.3 mmol/L ± 0.5 mmol/L, respectively) than in control group (18.9 mmol/L ± 0.9 mmol/L, 21.6 mmol/L ± 0.4 mmol/L, 22.5 mmol/L ± 0.7 mmol/L, all P<0.05) . (3) V/A ratio: Immediately after bypass and 12 hours postoperatively in the perfusion group (1.16±0.05, 1.20±0.05) was significantly lower than that in the control group (1.53±0.07, 1.68±0.25, all P<0.01). (4) OI: Immediately after bypass and 12 and 24 hours postoperatively in the perfusion group (370±33, 388+41, 414±40) was significantly higher than that in the control group (217±30, 210±36, 222±33, all P<0.05). (5) The time of mechanical assisted ventilation: Immediately after bypass and 12 and 24 hours postoperatively in the perfusion group (13h±4 h) was significantly lower than that in the control group (27h±6 h, P<0.01). Conclusion Pulmonary artery perfusion with washing red blood cell protective solution can alleviate lung injury after cardiopulmonary bypass.

Objective to investigate the effects of Celecoxib on the proliferation and early apoptosis of K562 cells and the expression changes of Rb protein (PRb) and P27Kip protein, and to observe whether the combination of Celecoxib and imatinib has a synergistic effect on the proliferation inhibition and apoptosis induction of K562 cells. Methods K562 cells were treated with different concentrations of Celecoxib (0, 10, 20, 40, 80 and 160 μ mol/L) for 36 h, and the cell viability was measured by tetramethylazolium salt colorimetric assay (MTT assay). Detection of early apoptosis in K562 cells by phospholipid binding protein V analysis; The effects of different concentrations of Celecoxib on the expression of PRb and P27Kip protein in K562 cells were detected by Western blot; Celecoxib was treated with 40 μ mol/L and 0.2 μ mol/L imatinib respectively or in combination to observe whether the drugs had synergistic effects on cell viability and apoptosis induction.Results Celecoxib could significantly inhibit cell viability in a drug dose-dependent manner: with the increase of Celecoxib concentration, the percentage of apoptosis of K562 cells increased correspondingly, and the apoptosis rate reached 25.92% at 160 μ mol/L concentration. Conclusion Celecoxib can inhibit the proliferation and induce apoptosis of K562 cells in a concentration-dependent manner. Its antiproliferative effect may be related to the down-regulation of PRb expression and the up-regulation of P27Kip expression. Celecoxib combined with imatinib has a significant synergistic effect on the proliferation of leukemia cells.

Objective to investigate the application value of video mediastinoscopy in the diagnosis of difficult mediastinal lesions.Methods 115 patients underwent video mediastinoscopy in our hospital were retrospectively analyzed.Among them, 102 patients underwent cervical mediastinoscopy, 13 patients underwent parasternal mediastinoscopy, and the operation was completed under general anesthesia or local anesthesia.91 patients had unknown diagnosis of simple mediastinal lesion.25 patients with high suspicion of lung cancer or clear non-small cell lung cancer with mediastinal lymph nodes.Results 37 patients were diagnosed as sarcoidosis, 14 cases of tuberculosis, 15 cases of lymphoma, 18 cases of unexplained mediastinal metastasis, and 6 cases of granuloma without caseous necrosis and typical "nodular cells". The diagnosis rate was 93.3% (84/90); The accuracy of diagnosis and staging of lung cancer was 100% (25/25) in 25 cases. The total diagnostic accuracy of video mediastinoscopy was 94.8% (109/115), and that of CT was 56.5% (65/115). The average operation time of this group was 26 minutes, and there were no surgical complications and mortality. Conclusion Video mediastinoscopy is safe and feasible for diagnosis and staging of lung cancer.

Objective to observe the effects of enteral immune nutrition (Ruineng and glutamine) on nutrition, immunity and acute inflammatory reaction in patients with gastrointestinal tumor during perioperative period.Methods 80 elderly patients with gastrointestinal tumor over 60 years old who underwent radical surgery were randomly divided into two groups:immune nutrition group (37 cases) (Ruineng and glutamine) (experimental group) and standard nutrition group (43 cases) (neng whole vegetarian) (control group). The two groups were treated with isocaloric enteral nutrition (125.4 kJ·kg-1·d-1). Intraoperative nasointestinal tubes or puncture jejunostomy (NCJ) were placed, and enteral nutrition was infused through the jejunum on the second day after operation for 7 consecutive days. Venous blood was collected on the 5th and 1st days before operation, and on the 1st and 9th days after operation. Serum albumin (ALB), prealbumin (PA), transferrin (TFN), T cell differentiation subset 3 (CD3), CD4, CD8, CD4/CD8, immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM) and C-reactive protein (CRP) were detected. Results On the 9th day after operation, serum TFN was 2.18g/L ± 0.29g/L, PA: 0.23g/L ± 0.09g/L, CD4:33.8%±5.4%, CD4/CD8 was 1.17±0.12, IgG: 13.2g/L ± 1.8g/L in the experimental group were significantly higher than those in the control group (P<0.05), while CRP was significantly decreased, from 41.3mg/L ± 19.0 mg/L in the control group to

Objective to investigate the characteristics of insulin slow pulse secretion patterns in normal glucose tolerance (NGT), overweight and abnormal glucose tolerance (IGT) individuals by using the technique of detection and analysis of insulin slow pulse secretion patterns. Methods 10 NGT individuals, 6 overweight individuals and 3 IGT individuals were treated with mixed meals. Blood was taken every 15 minutes for 24 hours, and blood glucose, insulin and C-peptide values were measured at each time point. Results (1) In mixed meals, slow insulin pulses appeared 2~4 after each meal, 3~4 at night, and 12~15 in the whole day. The first pulse occurred 30~60 min after meal, and the maximum pulse occurred 45~90 min after meal. This distribution characteristic was not statistically significant among normal control group, super recombination group and IGT group; (2) The mean pulse amplitudes of 24-h insulin secretion rate (ISR) in the three groups were 357 pmol/min±11pmol/min, 820 pmol/min±37 pmol/min, 666 pmol/min±53 pmol/min, respectively; (3) The average ISR pulse period was 75 min in the control group and 75~90 min in the super-recombination group, while the cycle rhythm was not obvious in the IGT group; (4) The cross-correlation functions of the three groups were 0.72±0.11, 0.80±0.11 and 0.51±0.11 respectively. Conclusion The characteristics of insulin slow pulse secretion pattern can be found by time series analysis.

Objective to observe the effect of dexamethasone (Dex) on the proliferation of human ovarian carcinoma cell line HO-8910 and to explore the role of small G protein RhoB signaling pathway in it. Methods the proliferation of human ovarian carcinoma cell line was detected by tetramethylazolium salt colorimetric assay (MTT assay) and soft agar assay. RhoB mRNA expression was detected by RT-PCR; Western blot was used to detect the expression of RhoB and its upstream and downstream signaling molecules phosphorylated Akt (p-Akt), p21cip1/waf1 and p27 proteins; The transcription of p21cip1/waf1 gene was detected by reporter gene technique.Results Dex could significantly inhibit the proliferation of HO-8910 cells in anchor-dependent and independent manner. The inhibition rate of the cells treated with 100 nM Dex for 6 days was 31%. The expression of RhoB mRNA and protein was significantly upregulated by 100 nM Dex (2.5 times that of the control). At the same time, the expression of p-Akt protein decreased (about 50% of control at 24 h), the expression of p27 protein increased (3.3 times higher than control at 36 h), and the transcription and protein expression of p21cip1/waf1 increased (about 1.7 times of control at 24 h). RhoB overexpression could decrease the growth rate of cells and enhance the proliferation inhibition of Dex (100 nM Dex treatment for 6 days, the inhibition rate was 44%); Conclusion the mechanism of Dex inhibiting the proliferation of HO-8910 cells may be related to the inhibition of PI3K/p-Akt signaling, thereby up-regulating RhoB protein and increasing the expression of downstream RhoB signaling molecules p21cip1/waf1 and p27.

Objective to investigate the effect of nuclear factor κ B "Decoy" oligonucleotide (NF-κ B Decoy ODN) on dextran sodium sulfate (DSS) colitis, and to provide experimental basis for finding a new drug for the treatment of ulcerative colitis (UC). Methods Thirty-six BALB/C mice were randomly divided into: normal control group; DSS + NF-κ B "bait" ODN group; DSS + mismatch ODN group; Methods (1) The concentrations of urea nitrogen, creatinine, random blood glucose, alanine aminotransferase and aspartate aminotransferase in serum of mice in each group were measured. (2) The colon tissues of mice were taken for gross and histomorphological observation. (3) The expression of NF-κ B p65 in colon mucosal cells was observed by immunohistochemical staining. (4) The content of tumor necrosis factor α (TNF-α) in colon mucosal tissues was determined by enzyme-linked immunosorbent assay (ELISA). (5) The distribution of NF-κ B "decoy" ODN in colon mucosal tissues of mice was observed by confocal microscope. Results (1) The DAI scores and histological scores of the mice treated with 5% DSS and the expression of TNF-α in intestinal mucosal tissues of the three groups were significantly higher than those of the normal control group (all P<0.05); However, the above indexes in the NF-κ B "bait" ODN group were significantly lower than those in the missense NF-κ B "bait" ODN group and the normal saline group (both P<0.01). (2) The expression of NF-κ B p65 in the intestinal mucosa of the three groups treated with 5% DSS was mainly in the nucleus, and there was no significant difference in the expression of NF-κ B p65 in the nucleus between the three groups. On the contrary, the expression of NF-κ B p65 in the normal control group was mainly in the cytoplasm. (3) NF-κ B bait ODN can effectively enter the colon mucosa and submucosa tissue of mice. (4) There was no significant difference in the liver, kidney function and blood glucose of mice in each experimental group. Conclusion NF-κ B "bait" ODN has a good protective effect on DSS colitis in mice.

Objective to investigate the relationship between the polymorphism of HindⅢ enzyme cleavage site in the non-recombinant region of Y chromosome and essential hypertension in Chinese Han nationality. Methods 857 male subjects were enrolled, including 402 patients with essential hypertension and 455 normal controls. All subjects were extracted from leukocyte DNA by conventional methods, and the polymorphism of HindⅢ enzyme cleavage site in the non-recombinant region of Y chromosome was detected by polymerase chain reaction combined with restriction endonuclease (HindⅢ) method. Results The genotypes of HindⅢ enzyme cleavage site in Y chromosome were significantly different between control group and essential hypertension group (P =0.02). The genotypes of HindⅢ (+) were 58.5% (235/402) in hypertensive patients and 66.4% (302/455) in normal population. The genotypes of HindⅢ (+) patients were less than those of normal people, and the genotypes of HindⅢ (-) patients were increased. Systolic blood pressure (133.8 mm Hg ± 25.2 mm Hg), diastolic blood pressure (83.5 mm Hg ± 13.3 mm Hg) and mean arterial pressure (100.2 mm Hg ± 16.5 mm Hg) of HindⅢ (+) patients The genotypes were significantly lower than those of HindⅢ (-) (respectively 138.0 mm Hg±27.0 mm Hg, 85.9 mm Hg±14.4 mm Hg, 103.3 mm Hg±17.6 mm Hg, all P<0.05) Conclusion HindⅢ polymorphism of Y chromosome is related to essential hypertension, which may be a genetic marker of essential hypertension in Chinese Han nationality.

Objective to investigate the relationship between baseline (1992) blood lipid level and 10 years (1992-2002) blood lipid change and carotid atherosclerosis in Peking University community cohort.Methods A cohort of 1985 people aged 35-64 years was established in Peking University community in 1992, and the baseline survey data were obtained; In 2002, the risk factors of cardiovascular disease were investigated again, and carotid artery ultrasonography was performed in this population. The relationship between baseline blood lipid level and 10-year blood lipid change and carotid atherosclerosis was analyzed in 1331 patients with complete data from the two surveys. Results (1) The thickening rate of carotid intima-media (IMT) was 47.8% and the incidence of carotid plaque was 29.9% in the study population aged 45-74 years in Peking University community. (2) When baseline total cholesterol (TC)<3.64mmol/L, the incidence of carotid plaque was 19.7% after 10 years, and when TC ≥6.24mmol/L, the risk of carotid plaque was 3.1 times (46.8%) that of the low TC level (<3.64mmol/L) group. With the increase of baseline TC level, the incidence of carotid plaque and carotid IMT thickening showed an increasing trend after 10 years (P<0.001). (3) Among the normal TC in 1992 and 2002, 319 patients (32.0%) had carotid plaque; Carotid plaque was found in 29 patients (50.0%) with abnormal TC in 1992 and 2002. Conclusion Carotid atherosclerosis was widespread in 45-74 years old. Both the incidence of carotid plaque and the thickening rate of IMT increased with the increase of baseline total cholesterol level.

Objective to investigate the relationship between the expression of nuclear transcription factor (NF-κ B) and apoptosis in testicular spermatogenic epithelial cells on the third day after 2 h testicular torsion reduction. Methods Twenty-four male SD rats were used to establish the left testicular torsion reduction model. Group Ⅱ animals were gavaged with equal amount of normal saline after testicular torsion and reduction; The expression of NF-κ B in testicular spermatogenic epithelial cells was detected in situ by Western Western blot and immunohistochemistry respectively. The apoptosis of spermatogenic epithelial cells was detected by TUNEL assay. Results the expression of NF-κ B protein (9.4±2.68) in the cytoplasm of spermatogenic epithelial cells of torsion testis in group Ⅱ was not significantly lower than that in group Ⅰ and Ⅲ (12±2.2; 11.1±3, respectively), and the expression of NF-κ B in the nucleus was significantly higher than that in group Ⅱ (8.4±3.1; 21.1±3.6; 6.0±2.3, respectively). The expression of NF-κ B in the spermatogenic epithelial cells of torsion testis in group Ⅰ and Ⅲ was mainly in the cytoplasm by immunohistochemistry in situ. The expression of NF-κ B in the spermatogenic epithelial cells of torsion testis in group Ⅱ was mainly in the nucleus, and the proportion of NF-κ B positive cells was significantly higher than that in the other two groups (15.6%±2.6%,66.1%±3.8%,10.8%±2.7%, respectively). The apoptosis level of spermatogenic epithelial cells (37.2%±3.3%) in group Ⅱ was significantly higher than that in the above two groups. Conclusion NF-κ B protein was activated and transferred from the cytoplasm to the nucleus on the third day after testicular torsion and reduction. The activation of NF-κ B protein was an important link in increasing apoptosis of spermatogenic epithelial cells, especially spermatogonia and spermatocytes.

Objective to investigate the clinical significance of detection of Y chromosome microdeletions in male infertility patients. Methods Two multiplex polymerase chain reaction techniques were used to detect Y chromosome microdeletions in 143 patients with severe oligozoospermia and azoospermia (80 cases with severe oligozoospermia and 63 cases with azoospermia) admitted to the First Affiliated Hospital of Nanjing Medical University from July 2003 to March 2005. Results Among the 143 patients, 21 (21/143, 14.7%) were found to have microdeletions, including 12 patients with idiopathic infertility (3 cases with severe oligozoospermia and 9 cases with azoospermia) and 9 patients with non-idiopathic infertility (3 cases with severe oligozoospermia and 6 cases with azoospermia). Among the 21 patients with microdeletions, 1 was located in AZFa region, 2 in AZFb + AZFc region, 2 in AZFb region and 16 in AZFc region. There was no significant difference in the location and extent of AZF microdeletion between idiopathic and non-idiopathic patients. Conclusion It is suggested that Y chromosome microdeletion should be detected in both idiopathic and non-idiopathic infertility patients, especially those who seek intracellular sperm injection therapy.

Objective to investigate the isolation and purification method of rat spermatogonia and the cellular characteristics after purification. Methods Testicular single cell suspension of 10 d-old SD rats was prepared by compound enzyme digestion, and spermatogonia was purified by differential adhesion combined with discontinuous Percoll density gradient centrifugation. The immunochemical characteristics of spermatogonia and purified cells in testis were observed with c-kit and α 6-integrin as cell markers, respectively. The positive expression rates of c-kit and α 6-integrin in purified cells were detected by flow cytometry, respectively. Results the spermatogonia expressed c-kit and α 6-integrin specifically in testis tissue, and the positive expression of c-kit and α 6-integrin was observed in purified single cells. The positive expression rates of c-kit and α 6-integrin were 1.59%±0.04% in unpurified group and 68.33%±2.45% in purified group, with significant difference between the two groups (P<0.01); The positive expression rate of α 6-integrin was 2.38%±0.60% in the unpurified group and 72.04% ±3.65% in the purified group, and the difference between the two groups was very significant (P<0.01). Trypan blue rejection test showed that the activity rate of purified cells was>95%. Conclusion C-kit and α 6-integrin can be used as surface markers of spermatogonia at a specific period.

ulcerative colitis (UC) is a non-specific inflammatory disease of rectum and colon with unknown etiology. At present, medical treatment is the main treatment, but 10% ~50% of patients still need surgical treatment. Total colorectal resection and ileal pouch anal anastomosis (IPAA) is an ideal surgical procedure for completely removing diseased tissue and reconstructing intestinal tract. However, the surgical operation is complicated and there are many controversial issues. In order to explore the therapeutic value of IPAA for UC, we treated 31 patients with ulcerative colitis with IPAA, and received good results.

In the past, people have always believed that RNA, as a relatively inert biological macromolecule, has no other role except to execute the instructions of DNA-building proteins.Recently, a series of studies have shown that small molecule RNA (microRNA, miRNA) actually manipulates many cellular functions, and can react to mRNA through the binding of complementary sequences, thus turning off or regulating the expression of related genes, and even regulating the development clock of cells through the switching of guide genes, which shows great potential in cutting-edge research fields such as stem cells and tumors [1]. This paper intends to explore its application prospect in miRNA research by combining liquid phase chip detection and analysis system.

H5N1 avian influenza virus can cause human lower respiratory tract infection and severe pneumonia, but so far, the type of cells to which it adheres in the lower respiratory tract of humans and laboratory animals is unclear. Although adhesion is not the only factor required for viral replication, this information is important for better understanding the pathogenesis of H5N1 avian influenza and screening suitable animal models. Debby van Rie1 and others from Erasmus Medical Center in the Netherlands conducted this study and published on 23 March 2006 in<Their latest findings are published on the website of the journal Science (http: / /www.sciencemag.org).

Antibiotic-resistant pneumococcal infections can cause treatment failure and increase medical expenses. Worldwide, the vast majority of antibiotic-resistant pneumococcal infections are caused by five serotypes, all of which are included in 7-valent pneumococcal homologous vaccines (6B, 9V, 14, 19F and 23F). It is precisely because of the relationship between serotypes and drug-resistant bacteria that homologous vaccines can be used to reduce the incidence of diseases caused by drug-resistant strains. In 2000, the United States approved the use of pneumococcal homologous vaccines, and it is recommended for infants under 2 years old and 2~4 years old with some chronic diseases or high risk factors. In order to evaluate the effect of vaccine on invasive diseases caused by drug-resistant strains, Moe H. Kyaw et al. conducted a study.

Parkinson's disease (PD) is an aging-related degenerative disease of the central nervous system. The main lesions are in the substantia nigra and striatum of the brain. The pathophysiological changes are the degeneration of dopaminergic neurons in the substantia nigra of the brain. The accumulation of oxidative DNA or RNA damage during aging is the main cause of inducing mutation and cell death. Oxidative DNA damage such as 8-oxyoguanine (8-oxoG) is significantly increased in the nucleus and mitochondrial genome of patients with various neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis.

Aortoiliac artery occlusive disease is a common vascular occlusive disease, and the incidence rate in China has been on the rise in recent years.From 1998 to 2003, 23 cases of aortoiliac artery occlusive disease were treated in Huashan Hospital affiliated to Fudan University, of which 17 cases underwent artificial vascular bypass grafting of aortofemoral arteries.

Interferon (IFN) -α is a cytokine with various biological effects, which plays an important role in anti-viral infection, tumor suppression and regulation of immune response [1]. It is also used in the treatment of many kinds of tumors and viral hepatitis in clinic, but its exact mechanism is not completely understood.

Breast cancer is one of the most common malignant tumors in women, and the incidence of breast cancer has been increasing in recent years.Previous research data at home and abroad have shown that the occurrence of breast cancer is related to gene alteration.Among them, the mutation of tumor suppressor gene p53 may be related to the malignant transformation of breast epithelial cells [1]. Recently, new members of p53 gene family-p51 and p73 genes have been discovered, which are located at chromosome 3q28 [2] and 1p36.3 respectively [3]. p73 and p51 genes have biological functions similar to p53 gene [1, 4, 5].

Prostate cancer (PCa) has become the first tumor disease to endanger men's health in European and American countries [1]. In recent years, the incidence of prostate cancer in China has also shown an obvious upward trend [2]. At present, the urgent clinical problem to be solved is the accurate diagnosis and timely treatment of early prostate cancer [3, 4]. In recent years, studies have confirmed that human gland kallikrein (hK2) may be a new method for diagnosing prostate cancer. We used real-time fluorescence quantitative polymerase chain reaction (PCR) to study the expression of hK2 in prostatic hyperplasia (BPH) and PCa tissue samples, and explored the specificity and clinical significance of hK2 in the diagnosis of prostate cancer.