Serum polyunsaturated fatty acid metabolites as useful tool for screening potential biomarker of colorectal cancer.

The biomarker identification of cancer is benefit for early detection and less invasion. Polyunsaturated fatty acid (PUFA) metabolite as inflammatory mediators can affect progression and treatment of cancer. In this work, the serum was collected from colorectal cancer patients and healthy volunteers, and then we tested the change of serum PUFA metabolites in both of them by ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Of the 158 PUFA and their metabolites, we found that abnormal change of 2, 3-dinor-8-iso-PGF2α, 19-HETE and 12-keto-LTB4 from arachidonic acid were observed in colorectal cancer patients. Meanwhile, 9-HODE and 13-HODE from linoleic acid were significant lower in colorectal cancer patients. Our data suggested that some PUFA metabolites might be used as a potential biomarker of colorectal cancer, which might provide assistance in clinical diagnosis and treatment.

Electroacupuncture ST36 prevents postoperative intra-abdominal adhesions formation.

BACKGROUND: We have recently proved electroacupuncture (EA) ST36 exerted an anti-inflammatory effect in the early phase of intra-abdominal adhesion formation. Evidences indicate that the anti-inflammatory effect of EA ST36 involves a cholinergic anti-inflammatory pathway-dependent mechanism via the vagus nerve. However, the exact effects and accurate vagal modulation of acupuncture in prevention of postoperative intra-abdominal adhesion formation has not been thoroughly evaluated. MATERIALS AND METHODS: Sprague-Dawley rats subjected to abdominal adhesion lesions operation at the cecum and abdominal wall were randomly divided into six groups as follows: (a) EAN: EA non-channel acupoints; (b) EA: EA ST36 after abdominal lesions; (c) VGX/EA: vagotomy (VGX) after abdominal lesions, then EA ST36; (d) VGX/EAN: VGX after abdominal lesions, then EAN; (e) α-BGT/EA: intraperitoneal injection of α-bungarotoxin (α-BGT, an antagonist of α7 subunit of cholinergic nicotinic receptor) before EA ST36, and (f) α-BGT/EAN group: α-BGT injection before EAN. Seven days after abdominal surgical lesions, the levels of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in the adhesive tissue were evaluated, macroscopic observation and histopathologic evaluation of adhesion formation and assessment of angiogenesis by immunohistochemical staining of platelet endothelial cell adhesion molecule-1 (CD31) were performed. RESULTS: EA ST36 reduced TNF-α and VEGF levels in adhesive tissue homogenates 7 d after surgery, whereas vagotomy or intraperitoneal injection of α-BGT before EA ST36 reversed its suppressive effects. EA at non-channel acupoints with or without vagotomy or intraperitoneal injection of α-BGT before EA had no suppressive effects on TNF-α and VEGF levels. EA ST36 alleviated the adhesion formation, with both of macroscopic and histopathologic adhesion scores significantly lower than those of the EAN group (1.56 ± 0.29 versus 3.00 ± 0.82, 1.35 ± 0.4 versus 3.91 ± 0.8, respectively, both P < 0.05). Compared with the EAN group, EA ST36 significantly decreased angiogenesis evidenced by reduced CD31 positive microvessel density in adhesive tissue. CONCLUSIONS: EA ST36 might reduce the postoperative local inflammatory response, attenuate the angiogenesis, and alleviate the adhesion formation partly via activating the cholinergic anti-inflammatory mechanism.

Reliable management of post-esophagectomy anastomotic fistula with endoscopic trans-fistula negative pressure drainage.

BACKGROUND: A gastroesophageal anastomotic fistula remains a potentially life-threatening post-esophagectomy complication. To promote fistula closure, we developed a modified endoscopic method of trans-fistula drainage with persistent negative pressure. In this study, we aimed to evaluate the efficacy of this endoscopic therapy. METHODS: Between June and November 2013, five male patients with post-surgical esophageal leakages who had undergone trans-fistula drainage therapy were treated with the modified endoscopic trans-fistula negative pressure drainage (E-TNPD) method. We placed a nasogastric silicone tube into the paraesophageal cavity through the fistula and accomplished drainage of the infected effusion with continuous negative pressure, resulting in shrinkage of the para-anastomotic cavity and eventual fistula closure. We withdrew the trans-fistula drainage when there were no signs of leakage, as confirmed by esophagography. Final closure was confirmed by esophagography before the patient was allowed to begin oral intake. RESULTS: E-TNPD was successful in all five patients. The median duration of drainage until tube removal was 34 days (range: 18 to 81 days). The duration for Cases 1 to 4 was 18 to 28 days. Case 5 suffered from multiple separate leaks at the anastomotic site and the gastric conduit. Complete restoration was achieved in 81 days for this patient. We found that in general, the earlier that trans-fistula drainage was established, the shorter the duration of hospitalization until complete defect closure. CONCLUSIONS: E-TNPD provided reliable and convenient management of post-surgical gastroesophageal anastomotic fistula and esophageal perforation. This method promoted fistula closure and prevented unnecessary repeated endoscopic examinations, extra equipment and expense.

Genetic landscape of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is one of the deadliest cancers. We performed exome sequencing on 113 tumor-normal pairs, yielding a mean of 82 non-silent mutations per tumor, and 8 cell lines. The mutational profile of ESCC closely resembles those of squamous cell carcinomas of other tissues but differs from that of esophageal adenocarcinoma. Genes involved in cell cycle and apoptosis regulation were mutated in 99% of cases by somatic alterations of TP53 (93%), CCND1 (33%), CDKN2A (20%), NFE2L2 (10%) and RB1 (9%). Histone modifier genes were frequently mutated, including KMT2D (also called MLL2; 19%), KMT2C (MLL3; 6%), KDM6A (7%), EP300 (10%) and CREBBP (6%). EP300 mutations were associated with poor survival. The Hippo and Notch pathways were dysregulated by mutations in FAT1, FAT2, FAT3 or FAT4 (27%) or AJUBA (JUB; 7%) and NOTCH1, NOTCH2 or NOTCH3 (22%) or FBXW7 (5%), respectively. These results define the mutational landscape of ESCC and highlight mutations in epigenetic modulators with prognostic and potentially therapeutic implications.

Si-RNA mediated knockdown of CELF1 gene suppressed the proliferation of human lung cancer cells.

BACKGROUND: Lung cancer is the leading cause of cancer-related death in the world, with metastasis as the main reason for the mortality. CELF1 is an RNA-binding protein controlling the post-transcriptional regulation of genes related to cell survival. As yet, there is little knowledge of CELF1 expression and biological function in lung cancer. This study investigated the expression levels of CELF1 in lung cancer tissues and the biological function of CELF1 in lung cancer cells. METHODS: CELF1 mRNA expression was determined in lung cancer and normal tissues, and the relationship between the expression level of CELF1 and clinicopathological parameters was evaluated. The biological function of CELF1 in A549 and H1299 lung cancer cell lines growth was examined. RESULTS: The expression of CELF1 was higher in human lung cancer tissues compared with the normal lung tissue. Lentiviral-mediated transfection of CELF1 siRNA effectively silenced the expression of CELF1 in both A549 and H1299 cells. Moreover, CELF1 knockdown markedly reduced the survival rate of lung cancer cells. Colony formation assays revealed a reduction in the number and size of lung cancer cell colonies from CELF1 knockdown. CONCLUSION: These results indicated that CELF1 may have significant roles in the progression of lung cancer, and suggested that siRNA mediated silencing of CELF1 could be an effective tool in lung cancer treatment.

Quantification of the sixth DNA base 5-hydroxymethylcytosine in colorectal cancer tissue and C-26 cell line.

BACKGROUND: DNA methylation at the five position of cytosine is well recognized as an important epigenetic modification in human health and disease. Recent evidences demonstrated that 5-methylcytosine (5-mC) by the TET family of enzymes can be converted to 5-hydroxymethylcytosine (5-hmC). Here, we use an ultrasensitive and accurate isotope-based LC-MS/MS method to precisely determine the levels of 5-hmC and 5-mC in colorectal cancer and the C-26 colon adenocarcinoma cell line. RESULTS: Our data showed that 5-hmC content is significantly reduced (approximately sixfold) in colorectal cancer as compared with adjacent normal tissue. Similarly, the ratio of 5-hmC to 5-mC dropped from 0.054 ± 0.005 in normal tissues, to 0.011 ± 0.002 in cancer. CONCLUSION: The analysis of 5-hmC levels and the ratio of 5-hmC:5-mC during tumor progression might provide insight into the role of this modification in cellular immortalization and transformation.

[Diagnostic accuracy research of needle puncture biopsy during operation for pulmonary single nodules].

OBJECTIVE: To investigate the diagnostic accuracy of needle puncture biopsy and pathological examination of frozen during operation for pulmonary nodules, and whether this diagnostic method can replace tumor resection examination. METHODS: Totally 50 patients (28 males and 22 females, average age was 59 years) who had the single nodule after imaging examination without any pathological diagnostic from January to October 2010 were selected in this research work. During open operation or video assisted thoracic surgery, needle (14 G model) was used to puncture biopsy for pathological examination of frozen. All the adverse events during puncture biopsy would be recorded. The resection specimens would be accepted paraffin pathological examination. The relationship between puncture frozen pathological and paraffin pathological examination was analyzed. RESULTS: All tumor sizes were ranged from 1.0 cm × 0.6 cm to 5.6 cm × 9.0 cm. The paraffin pathological examination after operation as the golden standard, there were 7 cases of benign tumor and 43 cases of malignant tumor. The diagnostic sensitivity of puncture biopsy was 90.7%, the specificity was 100%, the positive predictive value was 100% and the negative predictive value was 63.6%. There were 11 cases of benign tumor diagnosed by needle puncture biopsy, among which 4 cases were proved as malignant tumor by paraffin pathology, and the false negative rate was 9.3%. The main risk of puncture biopsy was bleeding after puncture immediately, and the rate was 4.0% (2/50). CONCLUSIONS: The puncture biopsy during operation had a high specificity for malignant lung tumor, and there was a certain false negative rate for benign tumor. Puncture biopsy and pathological examination of frozen tissue can replace tumor section biopsy in a way.

[Diagnostic value of enhanced computed tomography plus vascular endothelial growth factor C expression in hilar and mediastinal lymph node metastasis of non-small cell lung cancer].

OBJECTIVE: To explore the diagnostic value of preoperative enhanced computed tomography (CT) plus vascular endothelial growth factor C (VEGF-C) expression in hilar and mediastinal lymph nodes metastasis of non-small cell lung cancer. METHODS: A total of 87 patients with non-small cell lung cancer (NSCLC) received preoperative chest enhanced CT scans and underwent standard radical operation and systematic lymph node dissection. Pathologic examination was selected as the gold standard to determine lymph node metastasis. The immunohistochemical method was used to detect the expression of VEGF-C. The predicting values of chest enhanced CT, VEGF-C expression and their combination for the diagnosis of hilar and mediastinal lymph nodes metastasis were evaluated through comparing the sensitivity, specificity and accuracy. RESULTS: The sensitivity of CT scan was 75.0%, specificity 59.6% and accuracy 66.7%. The positive expression rate of VEGF-C was 78.2% (68/87) and strong positive rate 13.8% (12/87). The sensitivity of VEGF-C was 97.5%, specificity 38.3% and accuracy 65.5%. The combination of CT and VEGF-C had a better accuracy (74.7%) and the sensitivity and specificity were 80.0% and 70.2% respectively. CONCLUSION: Compared with CT scan or VEGF-C expression alone, the combination of CT and VEGF-C improves the specificity and accuracy of diagnosing lymph nodes metastasis in NSCLC. If this combination method is employed before therapy, the accuracy of clinical nodal staging may be enhanced.

[The effects of oral rehydration on ischemia injury in gut in 40% blood volume loss in rats].

OBJECTIVE: To investigate the effects of oral rehydration with glucose electrolyte solution(GES) on intestinal ischemia injury in 40% blood volume loss in rats. METHODS: SD rats were randomly divided into three groups (n=24): oral rehydration without hemorrhage (GES), hemorrhage without oral rehydration (HS), hemorrhage resuscitated with oral GES(HS + GES). About 4% of total blood volume was bled from the right common carotid artery of rats to produce a model of hemorrhagic shock. GES, which volume was three times of blood loss was given to GES group and HS + GES group in 0.5 h, 1 h and 6 h by a gastric tube post bleeding. The intestinal blood flow(IBF) were measured by laser Doppler at 2 h, 4 h and 24 h post hemorrhage. Animals were sacrificed, and specimens of intestinal tissue was taken for evaluation of Na+ -K+ -ATPase, diamine oxidase (DAO) and the rate of tissue water content, and assessment of the intestinal pathological changes. RESULTS: The IBF and the activity of Na+ -K+ -ATPase in HS+ GES group were dramatically higher than those in HS group (P < 0.05), and lower than those in GES group (P < 0.05). The water content of intestinal tissue in HS group were dramatically higher than those in GES group (P < 0.05), and lower than those at 2 h and 4 h, but dramatically higher than those at 24 h in HS + GES group. The activity of DAO at 24 h in HS+ GES group was higher than those in HS group (P < 0.05), and lower than those in GES group (P < 0.05). Less edema and hyperemia were found in HS + GES group than those in HS group at 24 h after bleeding. CONCLUSION: It is indicated that oral rehydration alleviate edema and ischemia injury in gut by increasing intestinal blood flow and the activity of Na+ -K+ -ATPase and DAO in the resuscitation of hemorrhagic shock.

[The protective effects of carbachol on visceral ischemia-induced lipid peroxidation injury in rats with sepsis].

OBJECTIVE: To investigate the effect of carbachol (CAR) on visceral perfusion and lipid oxidation injury in rats with sepsis. METHODS: Sixty-four Sprague-Dawley (SD) rats received cecal ligation and puncture (CLP) surgery, and they were divided randomly into two groups: septic model group (CLP group, n=32) and septic model with CAR-treatment group (CAR group, n=32). CAR (10 microg/kg, CAR group) or normal saline (CLP group) was immediately injected into penial vein. Sixteen animals in each group were used to observe the mortality rates 12 hours and 24 hours after CLP, and the remaining rats for measurement of variables of blood and tissue. At the 18 hours after CLP, the mean arterial pressure (MAP), the blood flow (BF) of liver, kidney and jejunum, the plasma levels of alanine aminotransferase (ALT) and creatinine (Cr) were measured. Animals were sacrificed after the aforementioned determinations, and specimens of liver, kidney and jejunum were harvested for evaluation of malondialdehyde (MDA), xanthine oxidase (XOD), and assessment of tissue water content (ratio of dry to wet weight) of those organs . The activity of diamine oxidase (DAO) in jejunal tissue was detected. RESULTS: The mortality rates of 12 hours and 24 hours of CAR group were 25.0% (4/16)and 50.0% (8/16) respectively, all significantly lower than those of CLP group [37.5% (6/16) and 75%(12/16), both P<0.05]. CAR treatment did not result in significant statistical difference in the levels of MAP compared with CLP group at 18 hours after CLP (P>0.05), but led to significant increases in BF of CAR group in liver, kidney and jejunum compared with those of CLP group (all P<0.05). The levels of XOD and MDA, as well as the tissue water content were significantly lower in CAR group than CLP group in kidney and jejunum (all P<0.05). The parameters of organ function were significantly different in CAR group compared with CLP group [ALT: (64.3+/- 8.3) U/L vs. (81.5+/-7.9) U/L, Cr: (96.4+/-7.0) micromol/L vs. (117.1+/-6.7) micromol/L, DAO: (0.20+/- 0.04) U/L vs. (0.12+/-0.03) U/L, all P<0.05]. CONCLUSION: The results indicate that CAR promotes visceral perfusion, inhibits lipid peroxidation production and alleviates visceral edema and dysfunction in rats with sepsis.

[The effects of electro-acupuncturing at Zusanli point on intestinal proinflammatory factors, diamine oxidase and tissue water content in rats with sepsis].

OBJECTIVE: To investigate the protective effect of electro-acupuncturing (EA) at Zusanli point on sepsis induced ischemic and oxygen free radical intestinal injury in rats with sepsis. METHODS: Thirty-two male Wistar rats were used to reproduce sepsis by cecal ligation and puncture (CLP), and they were randomly divided into four groups (each n=8): CLP+EA (CLP/EA), CLP+sham EA (CLP/SEA), vagotomy+CLP+SEA (VA/CLP/SEA) and vagotomy+CLP+EA (VA/CLP/EA). Zusanli point was electro-acupunctured with constant voltage (2-100 Hz,2 mA for 30 minutes) immediately after CLP surgery. Abdominal vagotomy was performed in rats in VA/CL/SEA and VA/CLP/SEA groups. Six hours after CLP, the mucosal blood flow of jejunum (JMBF) was measured. Animals were sacrificed after 6 hours and specimens of jejunum were harvested for evaluation of malondialdehyde (MDA), xanthine oxidase (XOD), diamine oxidase (DAO) and assessment of the water content (WCR). RESULTS: JMBF and the activity of DAO of CLP/EA group were markedly higher, and the levels of XOD, MDA and WCR in jejunal tissue were obviously lower than those of CLP/SEA group (all P<0.05). The levels of JMBF and DAO of the VA/CLP/SEA group and VA/CLP/EA group were significantly lower, and XOD, MDA and WCR obviously higher than those of the CLP/EA group ( all P<0.05 ). There were no statistically differences in all above measurements between the VA/CLP/EA group and the VA/CLP/SEA group (all P>0.05). CONCLUSION: The results indicate that EA at Zusanli point obviously increased JMBF and DAO, and alleviated tissue edema and insult of intestinal mucosa. Vagotomy could weaken or eliminate the effects of EA. It is suggested that cholinergic anti-inflammatory pathway is one of the main mechanisms of intestinal protective effect of EA at Zusanli point.

[Effects of pigment epithelial derived factor gene on growth of lung cancer cell and neovascularization: experiments with lung cancer cells and chick embryos].

OBJECTIVE: To investigate the effect of pigment epithelium derived factor (PEDF) on the growth of lung cancer cells and the cancer-related neovascularization. METHODS: The full length of human PEDF gene was amplified by polymerase chain reaction (PCR). Human lung cancer cells of the line SKEMS1 were cultured and transfected with PEDF(exp), An eukaryotic expression vector constructed by recombinant DNA technology, so as to construct the SKMES1(PEDFexp) cells over-expressing PEDF protein. RT-PCR and Western blotting were used to confirm the mRNA and protein expression of PEDF in these cells. Another SKMES1 lung cancer cells were transfected with blank plasmids (SKMES1(pEF/His) cells). The SKMES1 cells not transfected were called SKMES1(WT) cells. The 3 kinds of SKMES1 cells were inoculated in the chorio-allantoic membrane (CAM) of chick embryo hatched for 7 days respectively. The size and weigh of the tumor were measured. The vessels density was examined. RESULTS: The tumor volume of the SKMES1(PEDFexp) group was (0.10 +/- 0.05) cm(3), significantly smaller than that of the control group [(0.17 +/- 0.07) cm(3), P = 0.016], and the mass of the SKME(SPEDFexp) group was (0.008 +/- 0.004) mg, significantly smaller than that of the control group too [(0.024 +/- 0.009) mg, P = 0.006]. The amount of first class neo-vessels of the SKMES1(PEDFexp) group was (15 +/- 3), significantly fewer than that of the control group [(41 +/- 9), P < 0.001]. The amount of second class neo-vessels of the SKMES1(PEDFexp) group was (75 +/- 22), also significantly fewer than that of the control group [(175 +/- 39), P = 0.001]. CONCLUSION: Inhibiting the growth of lung cancer cells and neovascularization, PEDF protein may be used as a potential biological drug to treat lung cancer.

[Diagnosis and treatment of intralobar pulmonary sequestration].

OBJECTIVE: To summarize the diagnosis and surgical treatment of intralobar pulmonary sequestration (PS). METHODS: The clinical data of 7 cases of intralobar PS, 5 males and 2 females, aged 15 - 38, was collected and analyzed. Macroscopic and microscopic pathological findings were recorded. The expression of protein p53 and carcinoembryonic antigen (CEA) was evaluated immunohistochemically in 6 samples obtained from lobectomy. RESULTS: All 7 patients were admitted with major features of intermittent lung infection. Diagnosis was confirmed in all 7 cases before operation by contrast enhanced helical CT or MRI. All patients were treated with surgical excision, of which lobectomy was performed in 6 cases and wedge resection in 1 case. No surgical death was reported. All the aberrant systemic arteries arose from the thoracic aorta. The histological pictures showed polycystic lesion in sequestrated area with fibrosis formation and chronic inflammatory cell infiltration in the surrounding pulmonary stroma. Hyperplasia of epithelium occurred in some parts of the cystic lesions. Positive protein p53 staining and diffuse CEA expression were detected in all the 6 cases, showing stronger protein p53 staining in whose superficial layer of hyperplastic epithelium than in the basal layer. The normal bronchial epithelium was not stained with p53 or CEA. CONCLUSION: The diagnosis of intralobar PS can be confirmed by enhanced contrast helical CT with 3-dimensional reconstruction, a non-invasive method. Surgical intervention, such as lobectomy, can be applied after complete control of pulmonary infection. Aberrant accumulation of p53 protein and CEA expression in the cystic epithelium inside PS tissues show a relationship with chronic inflammation.

[A clinical study of thoracic-abdominal double-incision and two-field lymphadenectomy in treatment of esophagogastric junction cancer].

OBJECTIVE: To explore the best operation pattern of esophagogastric junction (EGJ) cancer and the regularity of lymph node metastasis in EGJ cancer according to Siewert typing. METHODS: Twenty-six patients with EGJ cancer received esophagogastrectomy by thoracic-abdominal double incision and two-field lymphadenectomy (12 cases) or by traditional left postero-lateral thoracotomy and lymph node sampling (14 cases). The outcomes were analyzed with SPSS 10.0 software RESULTS: (1) The number of lymph node dissection group of the thoracic-abdominal double incision group was 7.3 lymph node groups, significantly more than that of the traditional left postero-lateral thoracotomy group (3.5 lymph node group, P < 0.001). The number of proved metastatic lymph nodes of the thoracic-abdominal double incision group was 1.9 groups, significantly higher than that of the traditional left postero-lateral thoracotomy group (0.9 group, P = 0.013). The distance between the esophageal incisal edge and the tumor was 5.8 cm in the thoracic-abdominal double incision, longer than that in the traditional left thoracotomy group (5.1 cm). The diaphragm was not damaged in the double-incision group, thus the influence to respiration and circulation was decreased. (2) The abdominal metastasis of Siewert type I cancer was not severe, the cancer of type II might metastasize to abdominal or thoracic cavity, and the main metastatic site of type III cancer was abdominal cavity. CONCLUSION: Thoracic-abdominal double incision and two-field lymphadenectomy helps increase the radical resection rate of EGJ cancer and study the regularity of lymph node metastasis.

[Clinical analysis of 123 gastric cardia carcinoma patients treated with surgical operation].

BACKGROUND & OBJECTIVE: The prevalence of gastric cardia carcinoma is increasing in recent decades, necessitating further research on it. However, there are still debates on its clinical management. This study was to summarize our experiences in surgical treatment of gastric cardia carcinoma. METHODS: A total of 123 gastric cardia carcinoma patients, received surgical operation, were divided into 3 groups according to surgical approaches: 72 in thoracic group, 40 in abdominal group, and 11 in thoracoabdominal group. Clinical data, including preoperative examination, surgical approach, lymph node dissection, and postoperative pathology, of the patients were analyzed. RESULTS: Setting pathologic results as golden standard, the correct diagnosis rates of preoperative ultrasound for serosal involvement, lymph node metastasis, distal esophageal involvement, and others (including liver metastases, extended invasion, and ascites) were 71.2%, 62.2%, 47.8%, and 100%, respectively; those of CT were 78.6%, 72.7%, 51.9%, and 100%, respectively. Endoscopy could indicate the distance between tumor and incisor, and barium meal showed the relationship between tumor and diaphragm. The curative resection rate was 94.3% (116/123); among the 116 cases, 108 (93.1%) were adenocarcinoma, 2 were squamous cell carcinoma, 2 were adenosquamous carcinoma, 2 were atypical carcinoid, and 2 were carcinoid; 84 (72.4%) had abdominal lymph node metastases, 6 (7.1%) had thoracic lymph node metastases, and 40 (34.5%) had distal esophageal involvement. CONCLUSIONS: Preoperative abdominal ultrasound and thoracoabdominal CT scan are helpful in evaluating respectability of gastric cardia carcinoma. Endoscopy and barium meal may be helpful in deciding the surgical approach. Abdominal lymph node is the main route of lymphatic dissemination of gastric cardia carcinoma. The efficacies of the 3 surgical approaches are similar; each has its benefit. Surgical modalities should be carried out individually according to Siewert classification and patient's conditions.

[Abnormal expression of Thy-1 as a novel tumor marker in lung cancer and its prognostic significance].

OBJECTIVE: The purpose of this study was to investigate the expression of Thy-1 immunohistochemically in different lung tumors and its prognostic significance in non-small cell lung cancer (NSCLC) cases. We also evaluate relationship between Thy-1 and p53 expression status so as to find any clue about the mechanism. METHODS: In this study, we used anti-Thy-1/CD90 antibody to detect the expression pattern of Thy-1 in different lung tumor sections, which were embedded in paraffin blocks. The expressions of Thy-1 in 175 lung tissue cases, including different pathological types, were analyzed as tissue array form. We also detect expression status in 91 NSCLC among these cases and analyze the relationship between Thy-1 and p53. The relationship between Thy-1 expression and patients' survival was studied. RESULTS: We first found that anti-Thy-1 antibody can strongly stain a nuclear molecule in different type of lung cancer cells. Among lung cancer cases, 89 (56.7%) cases showed strong nuclear staining for Thy-1 specially. In univariate and multivariate analysis for 91 NSCLC patients we found TNM staging, lymph node status and Thy-1 overexpression in nuclei were independent factors to affect the prognosis of NSCLC patients. In lymph node non metastasis subgroup cases, Thy-1 negative patients had significant longer survival than Thy-1 positive cases (mean survival: 46.42 mons vs 38.56 mons, P = 0.0207). There was a significant association between Thy-1 and p53 expression (P < 0.0001). CONCLUSION: There is a significant overexpressed Thy-1 located in lung cancer cell nucleus as compared to the normal tissue or benign tumor cells of lung, and it is one of the factors effected on the prognosis of NSCLC patients. This finding suggests that Thy-1 maybe a novel latent malignant marker in the lung cancer pathology. The association between Thy-1 and p53 expression in nucleus suggests that p53 protein and Thy-1 may have some interaction.

[The expression of 11 cancer/testis (CT) antigen genes in esophageal carcinoma].

OBJECTIVE: To investigate the expression status of 11 different cancer/testis (CT) antigen genes in esophageal carcinoma. METHODS: Esophageal carcinoma tissue and adjacent normal esophageal mucosa taken from 35 esophageal carcinoma patients were assayed for the expression of 11 different CT antigen genes by RT-PCR techniques. RESULTS: Of the 11 CT antigen genes analyzed, none of them was expressed in normal esophageal mucosa. MAGE-3 was found to be the most frequently expressed in esophageal carcinoma tissues (62.9%), followed, in the order of expression frequency, by MAGE4 (31.4%), LAGE-1 (28.6%), MAGE-1 (25.7%), CT10 (20.0%), NY-ESO-1 (20.0%), CT7 (5.7%) and SCP1 (2.9%). No expression of SSX-1, SSX-2 and SSX-4 was found. Among the 35 cases, 28 (80.0%) expressed at least one CT antigen gene, 21 (60.0%) expressed more than 2 CT antigen genes, and 4 of the 21 (19.0%) expressed more than 4 CT antigens, which accounted for 11.4% of total number of patients (4/35). No CT antigen expression was found in the tumor tissue in 7 cases, including 5 cases in stage II and 1 case each in stage I and IV, respectively. Of the 11 CT genes examined, expression of 5 genes (NY-ESO-1, LAGE-1, MAGE-1, MAGE-3 and MAGE-4) was correlated with tumor progression. SCP-1 and CT10 expression was found more frequently in early stage patients. With progression of the disease, the frequency of co-expression of multiple CT antigen genes was significantly increased reaching 28.6% in stage III patients. CONCLUSION: Of the 11 different CT antigen genes examined by RT-PCR in esophageal carcinoma, 8 genes were detected in various frequencies in 28 of the 35 esophageal cancer patients studied. They are candidate tumor-associated antigens in the preparation of tumor vaccines for immunotherapy in esophageal cancer patients.

[Expression of placental growth factor and pigment epithelium-derived factor in non-small cell lung cancer and the prognostic significance thereof].

OBJECTIVE: To study the expression of placental growth factor (PlGF) and pigment epithelium-derived factor (PEDF) in non-small cell lung cancer, explore the possible role of these factors in the process of neovascular forming, and understand the predict function of these factors to verdict non-small cell lung cancer patients' prognosis. METHODS: Immunohistochemistry was used to detect the expression of PlGF and PEDF in 81 surgical specimens of non-small cell lung cancer from patients. CD31 immunohistochemical staining was used to measure the microvessel density (MVD) so as to investigate the microvessel forming status. The association of these two factors with MVD and their predictive function for the prognosis of non-small cell lung cancer were analyzed. RESULTS: The positive rates of PlGF and PEDF in the non-small cell lung cancer specimens were 43.2% (35/81) and 53.1% (43/81) respectively. There was a positive relationship between PlGF positive expression and MVD, and a negative relationship between PEDF positive expression and MVD. Furthermore, there was a negative correlation between PlGF and PEDF expression. PlGF was an independent prognostic factor in both univariate and multivariate analysis. CONCLUSION: PlGF and PEDF are expressed in non-small cell lung cancer. They are antagonistic to each other in the process of neovessels forming. PlGF plays a more important role in the progression of non-small cell lung cancer and prediction of its prognosis compared with PEDF.

[Expression of multiple cancer-testis antigen genes in non-small cell lung cancer treated by chemotherapy prior surgery].

OBJECTIVE: To investigate the possibility of utilizing cancer-testi (CT) antigens as targets for immunotherapy of non-small cell lung cancer (NSCLC) with vaccines. METHODS: Tissues from 51 NSCLC patients who had chemotherapy prior surgery were assayed for the expression of 11 different CT antigens by RT-PCR. RESULTS: Of the 11 CT antigens analyzed, MAGE-3 was found to be expressed most frequently in NSCLC tissues and CT7 the least frequently. The frequencies of CT antigen expression was: MAGE-3 (38%), NY-ESO-1 (21%), CT10 (17%), LAGE-1 (15%), MAGE-4 (13%), SCP-1, SSX1 and SSX4 (12%), MAGE-1 (10%), SSX2 (6%), and CT7 (2%). Among these cases, 34 (67%) expressed at least one CT antigen gene. 13 of the 17 cases in which no CT antigen expression was found in the tumor tissue, the tumors were classified as at the stage I. MAGE-3 and CT10 were found to be expressed more frequently in tissues from patients with late stage diseases while SCP-1 was found more frequently in earlier stages of NSCLC. CT expression was more frequently found in squamous cell carcinoma than in adenocarcinoma. CONCLUSIONS: (1) Cancer vaccines with CT antigens including MAGE-3, NY-ESO-1, LAGE-1, etc, are suitable for immunotherapy of NSCLC after chemotherapy and surgery. (2) NSCLC patients at different stages of disease may be treated with vaccines of different CT antigen composition. (3) CT antigen vaccines are most attractive for patients with late stage NSCLC and/or squamous cell carcinoma of NSCLC.

[Detection of human papillomavirus type 16 E6 mRNA in carcinomas of upper digestive tract].

OBJECTIVE: To investigate the relationship between human papillomavirus (HPV) infection and the development of upper digestive tract carcinomas. METHODS: In situ hybridization (ISH) technique was used to detect the expression of HPV16 E6 mRNA in 40 specimens of esophageal squamous cell carcinoma, 74 specimens of gastric cardiac adenocarcinoma, 40 specimens of adenocarcinoma of gastric corpus, 62 specimens of adenocarcinoma of gastric antrum, all resected during operation, and 58 specimens of normal tissues of gastric mucosa, obtained by endoscopy. RESULTS: The positive rate of HPV16 E6 mRNA was 70% (28/40) in esophageal squamous cell carcinomas, 67.6% (50/74) in gastric cardiac adenocarcinomas, 47.5% (19/40) in adenocarcinoma of gastric corpus, 35.5% (22/62) in adenocarcinoma of gastric antrum, and 20% (10/50) in normal tissues of gastric mucosa respectively, without a significant difference between esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma (P = 0.955). The positive rate in gastric adenocarcinoma was significantly lower than that in gastric cardiac adenocarcinoma (P < 0.05) or in esophageal squamous cell carcinomas (P < 0.01). The positive rate in normal tissue of gastric mucosa was significantly lower than those in gastric adenocarcinoma (P < 0.05) in gastric cardiac adenocarcinoma (P < 0.01), and in esophageal squamous cell carcinoma (P < 0.01). The positive rate in gastric cardiac carcinomas was negatively correlated with TNM stage (P < 0.05). However, there was no correlation between HPV16 and gender, age, tumor size, depth of penetration, differentiation, lymph node metastasis, vascular invasion and prognosis of these patients with gastric cardiac carcinoma (all P > 0.05). CONCLUSION: HPV infects not only squamous epithelium, but also glandular epithelium. HPV16 may be an etiologic factor in the development of upper digestive tract carcinomas, especially esophageal and cardiac carcinomas.