Volume 86  ·   Issue 22  ·  2006

Total mesorectal excision (TME) and extended radical excision (TME) have been widely used in the treatment of rectal cancer, but there are also great controversies. We analyzed the similarities and differences between them and discussed them with our colleagues.

Objective to construct a small interfering RNA (siRNA) expression vector targeting human MAGE3 gene, and to observe the effect of RNA interference-mediated expression of MAGE3 gene on colonization and metastasis of lung cancer cells. Methods A hairpin-shaped siRNA targeting MAGE3 was designed, and two complementary oligonucleotide strands were synthesized according to the design, annealed and ligated into pSUPERneoGFP vector. After transformation and amplification, the sequence was determined. Human lung cancer cell NCI-H446 was transfected with liposome encapsulation, and the expression levels of mRNA and protein of MAGE3 gene were detected by RT-PCR and Western blot. The effects on tumor formation and invasion were observed by soft agar cloning experiment and cell invasion experiment in vitro. Results The double-stranded oligonucleotide fragment of siRNA targeting MAGE3 gene was cloned into pSUPERneoGFP vector, and the results were correctly identified and sequenced by enzyme digestion; RT-PCR and Western blot showed that the expression level of MAGE3 gene was significantly reduced; The number of clones formed in soft agar and the number of cells penetrating Matrigel in lung cancer cells transfected with siRNA expression vector decreased significantly, compared with those in untransfected and empty vector groups (P<0.05). Conclusion The siRNA vector targeting MAGE3 gene was successfully constructed, and the silencing of MAGE3 gene expression mediated by RNA interference can effectively reduce the colonization and metastasis of lung cancer cells.

Objective to investigate the correlation between the biological behavior of breast cancer and the above three indexes by detecting the expressions of thymidylate synthase (TS), thymidine deoxynucleoside phosphorylase (TP) and difluoropyrimidine dehydrogenase (DPD) in breast cancer samples. Methods Immunohistochemistry (ABC) was used to detect the expressions of TS, TP and DPD in 228 breast cancer tissues. Results TS, TP were significantly correlated with tumor size, lymph node status, histological grade and MVC, but not with age and hormone receptor status (ER, PR) of patients. There was no correlation between the expression of DPD and many clinicopathological factors such as age, tumor size, lymph node status, hormone receptor status (ER, PR), histological grade and MVC. The 10-year disease-free survival rate and the overall survival rate of patients with positive TS and TP expression were significantly lower than those with negative expression (0, 0, 3.9%, 0.9% and 25%, 25.4%, 58.8% and 61.8%), with significant statistical differences (all P<0.01). The 10-year disease-free survival rate and the overall survival rate of patients with positive and negative DPD expression were 8.8%, 16.2%, 23.3% and 39.5%, with no significant differences (all P<0.05). Multivariate analysis showed that the histological grade, lymph node status, TS and TP expression were significantly correlated with the overall survival rate of patients, and the risk coefficients of TS and TP were 0.379 and 0.219, respectively, both of which were less than 1, which also indicated that both of them were very strong protective factors for the overall survival rate. Conclusion The level of TS and TP expression may be an important prognostic index of breast cancer patients; There was no correlation between the expression of DPD and clinicopathological factors, 10-year disease-free survival and overall survival.

Objective to detect and analyze the distribution of single nucleotide polymorphism (SNP) loci in patients with trunk conus malformation and normal population.Methods the genotypes of three SNP loci in 130 patients with trunk conus malformation and 200 normal subjects were analyzed by polymerase chain reaction-restriction fragment length polymorphism method. The genotypes and allele frequencies of SNP loci in patients and control groups were statistically analyzed by contingency table method; The haplotypes were constructed by PHASE software and statistically analyzed whether there was any difference in haplotype frequency between the patient group and the control group.Results The distribution of allele frequency and genotype frequency of G2963A locus was significantly different between the patient group and the control group.The frequency of G allele in the patient group was significantly higher than that in the control group (χ 2=8.14, P<0.005). The haplotype analysis showed that the distribution frequency of four haplotypes between the patient group and the control group was statistically significant (χ 2=22.39, P<0.005): G2857/G2963/A6571 and G2857/G2963/T6571 were common haplotypes in the population. The frequency of G2857/G2963/A6571 and C2857/A2963/T6571 haplotypes in the patient group was higher than that in the control group.Conclusion The SNP locus G2963A in the coding region of TBX1 gene is significantly correlated with trunk conus malformation, and the risk of trunk conus malformation in people with G allele is relatively higher; The haplotype of the three SNP loci has some significance, and may be linked to the susceptibility gene of conic trunk malformation.

Objective to investigate the feasibility of microsurgical biliary tract reconstruction to reduce the incidence of biliary tract complications after living donor liver transplantation (LDLT). Methods Thirty-two patients with LDLT (8 children and 24 adults) who were treated in our hospital from January 2001 to December 2005 were analyzed. There were 43 hepatic duct openings in 32 donor liver grafts (24 in right liver, 6 in left liver and 2 in left lateral lobe). Therefore, a total of 43 hepatic duct openings were treated for biliary tract reconstruction. Microsurgical technique was used for biliary tract reconstruction. Among them, 21 anastomoses were anastomosed by end-to-end bile duct and 13 anastomoses of 22 hepatojejunostomies were anastomosed by surgical magnifying glass, and 9 hepatojejunostomies were anastomosed by surgical microscope. Results Two recipients had biliary tract complications after surgery. One patient underwent hepatojejunostomy under surgical magnifying glass, and the anastomotic leakage occurred on the third day after surgery, which was cured after open drainage again, and the other patient developed mild anastomotic stricture of the biliary duct at 3 months after surgery. Conclusion Microsurgical technique of biliary tract reconstruction in living donor liver transplantation, especially the anastomosis under surgical microscope for hepatic duct jejunostomy of very thin hepatic duct (diameter ≤2 mm), is helpful to reduce the incidence of biliary tract complications after LDLT.

Objective to explore the feasibility of screening sleep apnea hypopnea syndrome (SAHS) by using polysomnography (PSG) as a control. Methods From February 2004 to November 2004, 80 patients in sleep centers were monitored by polysomnography (PSG) all night (more than 7 h), and simultaneously performed dynamic electrocardiogram examination. Both parties calculated the sleep apnea hypopnea index (AHI) separately without communication with each other, and made positive and negative diagnosis of sleep apnea hypopnea syndrome. The PSG results were used as the gold standard to evaluate the feasibility of screening SAHS by using dynamic electrocardiogram EDR technique. Results Software was used to analyze automatically, and the sensitivity of software was set to 75%, 87.5% and 100%, respectively. When the sensitivity was 75%, the sensitivity of EDR to detect SAHS was 26.7%, the specificity was 80%, the positive predictive value was 80%, the negative predictive value was 26.7%, and the diagnostic coincidence rate was 40%; When the sensitivity was 87.5%, the sensitivity was 55%, the specificity was 45%, the positive predictive value was 75%, the negative predictive value was 25%, and the diagnostic coincidence rate was 52.5%; When the sensitivity is 100%, the sensitivity is 88.3%, the specificity is 35%, the positive predictive value is 84.1%, the negative predictive value is 50%, and the diagnostic coincidence rate is 75%. Conclusion EDR technique of dynamic electrocardiogram and automatic analysis of software can be used for the primary screening of SAHS patients, among which the sensitivity and coincidence rate of automatic analysis are higher when the sensitivity is 100%.

Objective to investigate the effects of α 1 antitrypsin (AAT) and lipopolysaccharide (LPS) on the synthesis of AAT in renal tubular epithelial cells. Methods the expression of AAT mRNA and protein in human renal tubular epithelial cell line (HKC) was detected by indirect immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR), and the PCR products were identified by DNA sequencing. HKC was stimulated with different concentrations of LPS, and divided into 4 groups according to the LPS concentration experiment. The changes of AAT mRNA and protein expression in HKC after LPS stimulation were detected by real-time fluorescence quantitative PCR and Western blot, respectively. Results the indirect immunofluorescence showed that HKC AAT was positive and distributed in the cytoplasm; RT-PCR detection found that HKC expressed AAT mRNA; Compared with the unstimulated group, real-time fluorescence quantitative PCR showed that the expression of AAT mRNA was significantly upregulated when stimulated by 2.0 μ g/ml LPS for 4 h (the fluorescence intensity ratio was 3.43±0.88 vs 1.22±0.20; P<0.01), and Western blot showed that the expression of AAT protein was significantly increased when stimulated by 2.0 μ g/ml LPS for 8 h (the band density ratio was 0.88±0.12 vs 0.59±0.05; P<0.01). However, 0.5 μ g/ml and 1.0 μ g/ml LPS did not significantly stimulate the expression of AAT mRNA and protein in HKC. Conclusion HKC can synthesize AAT, and LPS can upregulate the expression of AAT mRNA and protein in human renal proximal tubular epithelial cells.

Objective to construct recombinant pshRNA-DNMT1 silencing DNMT1 gene and investigate its effect on the proliferation and apoptosis of gastric cancer AGS cells in vitro and in vitro.Methods A short hairpin RNA (shRNA) oligonucleotide fragment was designed and cloned into vector pTZU6+1. Recombinant pshRNA-DNMT1 was constructed and transfected into gastric cancer cell line AGS. The protein level of DNMT1 was detected by Western blot, the mRNA level was assessed by reverse transcription-polymerase chain reaction (RT-PCR), the number of viable cells was dynamically monitored by tetramethylazole salt colorimetric assay (MTT assay), and its pro-apoptotic effect in vitro was observed by electron microscope and in situ end labeling (TUNEL). The cell apoptosis was detected by hematoxylin-eosin (HE) staining, electron microscopy and TUNEL, and the proliferation ability was evaluated by proliferating cell nuclear antigen (PCNA) assay. Results the recombinant plasmid pshRNA-DNMT1 was successfully constructed and confirmed by sequence analysis. The expression of DNMT1 protein decreased at 24 h, 68.54% at 48 h and 81.47% at 72 h after transfection with pshRNA-DNMT1. The inhibition rate of pshRNA-DNMT1 was 21.63% at 24 h, 52.97% at 48 h and 72.06% at 72 h after transfection. The cell survival rate at 24, 48 and 72 h after transfection was 79.49%, 51.63% and 39.16% in PBS group. The recombinant plasmid pshRNA-DNMT1 can inhibit tumor growth in a time and dose dependent manner. Conclusion the recombinant plasmid pshRNA-DNMT1 can specifically and effectively inhibit the tumor growth of gastric cancer cell line AGS

Objective to investigate the prevalence of peripheral vascular disease (PVD) and its main risk factors in people with diabetes mellitus and impaired glucose regulation (IGR) in Shanghai community. Methods 717 patients with diabetes mellitus and IGR obtained from the epidemiological survey of metabolic syndrome in Caoyang community of Shanghai from 2000 to 2001 were followed up for 5 years. Among them, 427 patients with complete follow-up and complete data were included in this analysis, including 210 males and 217 females, aged 21-104 years (67.3 years ± 14.0 years). PVD was diagnosed by using Nicolet Versalab SE two-way Doppler flow detector to detect the ankle/brachial artery pressure ratio (A/BI), and intermittent claudication was investigated by Edinburgh claudication questionnaire. The influencing factors related to PVD were analyzed by Logistic regression. Results The prevalence of PVD in hyperglycemia population in Caoyang community was 12.2%, of which 15.1% in diabetic population and 7.7% in IGR population. Only 13.5% of patients with ABI ≤0.9 had intermittent claudication symptoms. Logistic regression analysis found that, Conclusion the prevalence of PVD in Shanghai community hyperglycemic population is high, not only in the stage of diabetes, but also in the pre-diabetic stage.Most patients with PVD have no clinical symptoms, so routine Doppler ABI detection in diabetes clinic is helpful for early diagnosis and early intervention of PVD.

Objective to investigate the expressions of hypoxia-inducible factor-1 α (HIF-1 α), HIF-2 α, vascular endothelial growth factor (VEGF) and microvessel density (MVD) in renal clear cell carcinoma, and to analyze their relationship and significance. Methods Immunohistochemical method was used to analyze the expressions of HIF-1 α, HIF-2 α, VEGF and MVD in 107 cases of renal clear cell carcinoma. Results The positive staining rate of HIF-2 α in this group (80.5%) was significantly higher than that of HIF-1 α (69.2%) (P<0.01). The positive rate of VEGF in HIF-1 α and HIF-2 α positive groups (90.5% and 89.8%) was significantly higher than that in their negative groups (51.5% and 26.3%), P<0.01. There was a positive correlation between the expressions of HIF-2 α and VEGF and MVD, (Spearman's correlation coefficient was 0.500 and 0.513, respectively), P<0.01. However, there was no correlation between the pathological grade and stage of tumor and the expression of HIF-1 α and 2 α, P<0.05. Conclusion HIF-2 α in renal clear cell

Objective to observe the frequency of CD4+ CD25 high regulatory T cells (Treg) in peripheral blood of patients with different types of HBV infection and the correlation of clinical indexes. Methods The peripheral blood of 72 patients with chronic hepatitis B (CHB), 16 patients with acute hepatitis B (AHB) and 32 healthy subjects was collected. The frequency of Treg and the expression of Treg cells on the surface and intracellular cells were analyzed by flow cytometry. All cases and controls were detected by enzyme-linked immunosorbent assay (ELISA), HBsAg, HBsAb, HBcAb, real-time fluorescence quantitative RT-PCR, and liver function was measured. Results CD4+ CD25 high Treg expressed high CD45RO, low CD45RA, and high CTLA-4 in cells. The frequency of Treg in CD4+ T cells was compared with that of chronic hepatitis B group (3.9% ± 1.4%) and normal controls (3.5% ± 0.7%). The difference was not statistically significant (P>0.05), but it was significantly higher than that of acute hepatitis B group (3.9% ± 1.4%) The frequency of Treg was significantly higher in the recovery phase (6.0% ± 1.7%) than in the acute phase (3.0% ± 0.6%) in 8 follow-up patients with acute hepatitis B, and the paired t-test was P<0.001. Conclusion CD4+ CD25 high Treg was significantly higher in the high viral load group of chronic hepatitis B patients, which suggests that Treg may affect virus clearance by inhibiting cellular immune response; Treg is low in the early stage of acute hepatitis and then increased, which may be related to the role of Treg in different stages of acute hepatitis.

The mechanism of hypertrophic scar formation has not been clearly revealed. In the past, the research on scar mainly focused on fibroblasts, but the research on vascular endothelial cells, another important cell component in scar, has been neglected. At present, it is believed that vascular endothelial cells are not only related to the integrity of microvascular endothelium, but also can secrete a large number of cytokines and hormones, such as fibroblast growth factor (FGF), metastatic growth factor (TGF) β 1, nitric oxide (NO), endothelin (ET) 1, etc., which are involved in the pathogenesis of many organs and systems.

A 37-year-old female was admitted to hospital with abdominal pain and diarrhea for 2 days. Two days ago, the patient developed persistent abdominal pain with diarrhea without obvious inducement, 7-8 times a day, dilute black stools, no fever, chills, and no obvious weight loss. The patient was admitted to hospital with "acute diffuse peritonitis". Two months ago, the patient was admitted to a local hospital with abdominal pain for 1 day. The patient was diagnosed as "acute peritonitis" and underwent exploratory laparotomy. During the operation, "omentum ischemic necrosis, yellow-white lesions with a diameter of 5-10 mm scattered on the intestinal surface" was found. Part of the omentum was removed, and the patient recovered smoothly after operation.

Compared with invasive ventilation, noninvasive ventilation has the advantages of convenient use, lower cost, alleviating patients' pain and fewer complications, etc. It has been gradually recognized by clinicians, and its clinical use is more and more widespread.There has been sufficient evidence to show that the use of non-invasive positive pressure ventilation (NIPPV), including continuous positive airway pressure (CPAP) or biLevel ventilation (biLevel), can reduce the secondary invasive mechanical ventilation in patients with cardiogenic pulmonary edema, but there is no evidence-based evidence of whether patients have other benefits.

metachromatic leukodystrophy (MLD) is a genetic disease that affects the expression of acyl sulfatase A (ARSA). It is characterized by oligodendrocyte (OL) dysfunction, cerebroside sulfate aggregation, demyelination and progressive neurodegeneration.

intestinal ischemia-reperfusion (IIR) is very sensitive to the injury of intestinal ischemia-reperfusion [1]. Many factors such as acute blood loss, shock and severe multiple trauma can cause the phenomenon of intestinal ischemia-reperfusion. intestinal ischemia-reperfusion not only causes the injury of intestinal tract [2], but also causes the injury of distant organs, and in severe cases can lead to multiple organ dysfunction syndrome [3]. sodium ferulate (SF) is one of the effective ingredients of traditional Chinese medicines such as Ligusticum chuanxiong and Angelica sinensis, which can be synthesized artificially.

Glutamate is one of the cochlear afferent neurotransmitters [1, 2], but recent studies have shown that glutamate has excitatory cytotoxic effects on various central and peripheral nerve cells, and studies have shown that magnesium ions have protective effects on the damage of central nervous system. However, does magnesium ions also have protective effects on spiral ganglion neurons cultured in vitro? The effect of glutamate on the damage of cochlear spiral ganglion cells in vitro and the effect of magnesium ion on this effect were observed. The effect of magnesium ion and glutamate on the concentration of calcium ion in cochlear spiral ganglion cells at rest was also observed, in order to provide some new strategies for the treatment of sensorineural deafness in the future.

The research points out that the increased risk of cardiovascular disease in the elderly is the result of the interaction between aging and disease [1,2]. The arrival of the global aging society makes the research of vascular aging an urgent social demand. The characteristic remodeling of vascular structure and function with aging is called vascular aging, and vascular remodeling caused by aging may become the target of cardiovascular disease treatment and prevention [3].