Objective:To explore the clinical significance of combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood for the diagnosis of colorectal cancer. Methods The data of patients admitted to the Department of Gastroenterology, Shanghai East Hospital Affiliated to Tongji University from January to September 2019 were retrospectively analyzed. They were divided into colorectal cancer group (62 cases of colon cancer, 59 cases of rectal cancer), precancerous lesions group (77 cases of colorectal adenoma, 5 cases of high-grade intraepithelial neoplasia), interference group (61 cases of colorectal cancer and advanced adenoma negative but suffered other intestinal lesions, 17 cases of non-colorectal cancer) and healthy group (94 cases). The methylation status of three genes (Septin9, SDC2 and BCAT1) in peripheral blood plasma was detected simultaneously by fluorescence PCR. The relationship between the positive rate of three genes detected jointly and the clinic pathological characteristics of colorectal cancer was analyzed and compared with serum carcinoembryonic antigen (CEA) positive rate. The colorectal cancer group was divided into stage Ⅰ, Ⅱ, Ⅲ and Ⅳ according to TNM stage, and the colorectal cancer group was analyzed and counted by grade. The diagnostic efficiency of detection methods was analyzed by receiver operating characteristic (ROC) curve, and the area under ROC curve (AUC) was compared.Results:The positive rate of combined detection of SDC2 and BCAT1 gene methylation was higher than other three groups (χ 2 =237.246, P<0.001). The positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation was higher than CEA in colorectal cancer group ( P<0.001). The positive rates of the combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅳ of colorectal cancer group were 73%(16/22), 87%(34/39), 86%(30/35) and 96%(24/25), respectively. Compared with CEA group, the positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅲ of colorectal cancer group was higher than serum CEA ( P<0.001), but the positive rate of stage Ⅳ was not statistically significant compared with CEA group ( P>0.05). ROC curve analysis showed that the AUC of Septin9, SDC2 and BCAT1 was 0.857(95% CI 0.810-0.903),0.819(95% CI 0.768-0.871)and 0.862(95% CI 0.816-0.909), respectively. The AUC of combined detection of three gene methylations was 0.889 (95% CI 0.846-0.933), and the AUC of combined detection with serum CEA was 0.913 (95% CI 0.874-0.951). There was no significant difference in the positive rate of combined detection of Septin9, SDC2 and BCAT1 gene methylation among different gender, age and cancerous site of colon cancer patients (all P>0.05). Conclusion:The combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood plasma is helpful for the early diagnosis of colorectal cancer. The positive rate in stage Ⅰ-Ⅲ of colorectal cancer group is higher than serum CEA. The combined diagnosis of the three genes can improve the diagnostic efficiency.

OBJECTIVETo explore the differences between conservative and surgical treatment for thoracic and lumbar spine tuberculosis on indications and clinical results.

METHODSFrom May 2000 to June 2008, 162 cases of thoracic spinal tuberculosis patients were studied retrospectively, including 89 males and 73 females, aged from 4 to 71 years (means 43.6 years). Among them, 38 cases of onset time was short, X-ray showed a narrow cone gap and MRI showed vertebral body signal changes, the ones who had better general condition were applicated of anti-TB drugs [6HREZ (S)/6-9HRE] and traditional Chinese medicine dialectical; 51 cases with thick swollen and vertebral body marginal damage but not affect the stability of spine, were treated with spinal tuberculosis debridement surgery; 73 cases with abscess, vertebral destruction of center, spinal instability or spinal cord function associated with damage to persons, were treated with surgical debridement and interbody bone grafting and fixation. Clinical observation were evaluated according to the standard cure for bone tuberculosis.

RESULTSThirty-eight cases by conservative treatment, had been cured in 1 to 2 years. Focus of infection cleared in 51 cases, 3 cases occurrenced the sinus next to incision, 4 cases of kyphosis angle (Cobb) lost 5 degrees-10 degrees compared with that before treatment (means 4.5 degrees). Seventy-three cases by internal fixation obtained neurological deficiency symptoms recovery; 2 occurenced the sinus next to the incision, 3 cases appeared subcutaneous emphysema of iliac area and local pain, but need no special treatment. A total of 136 patients were followed-up from 12 to 60 months with an average of 32.8 months, and had been all clinically cured.

CONCLUSIONPatients with early detection only on the imaging showing vertebral lesions, without obvious sequestrum, abscesses, can be selected for conservative anti-tuberculosis treatment. Patients combined with abscess, vertebral destruction on light degree and not affected the stability of the spine, can be removed by simple surgery to obtain better efficacy. Patients with abscesses, sequestrum, spinal vertebral instability leading to heavy damage associated with spinal cord or nerve function impairment,will need surgical removal of lesions of tuberculosis, give graft and spinal fixation at the same time.

Adolescent ; Adult ; Aged ; Antitubercular Agents ; therapeutic use ; Child ; Child, Preschool ; Female ; Humans ; Lumbar Vertebrae ; drug effects ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Vertebrae ; drug effects ; surgery ; Treatment Outcome ; Tuberculosis, Spinal ; drug therapy ; surgery ; Young Adult

AIMTo investigate the effect of ciliary neurotrophic factor (CNTF) on the nuclear translocation of protein kinase C (PKC) following NMDA administration in the primary cultured hippocampal neurons.

METHODS(1) PKCGAMMA or PKCepsilon- immunocytochemistry staining method was used after treating neurons with NMDA or CNTF. (2) The gray of the nucleus of the PKC-positive neurons were measured under the image pattern analysis system.

RESULTS(1) After NMDA administration of different concentration and time, Nucleus appear PKCgamma and PKCepsilon activities, especially the 100 micromol/L NMDA 30 min group. (2) The gray of nucleus in CNTF + 500 micromol/L NMDA group is similar to control group.

CONCLUSIONNMDA can induce nuclear translocation of PKC in the primary cultured hippocampal neurons, and CNTF can inhibit the translocation. It suggests that the inhibition of PKC translocation induced by NMDA is one of the important reasons for the neuro-protective effects of CNTF.

Animals ; Cells, Cultured ; Ciliary Neurotrophic Factor ; pharmacology ; Hippocampus ; cytology ; N-Methylaspartate ; pharmacology ; Neurons ; drug effects ; metabolism ; Protein Kinase C ; metabolism ; Protein Transport ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of down-regulating Tim-3 gene in the peripheral blood mononuclear cells (PBMCs) of an asthmatic mouse model by short hairpin RNA (shRNA) and to explore the effect of Tim-3 on Th1 and Th17 cell differentiation.

METHODSAn asthmatic murine model was established by ovalbumin sensitization and challenge. PBMCs were isolated from asthmatic mice and transfected by shRNA targeting Tim-3 gene. The mRNA and protein expressions of Tim-3 were detected by quantitative PCR and Western blot. Flow cytometry analysis was performed to determine the levels of Th1 and Th17, and ELISA was performed to determine concentrations of IFN-γ, IL-4 and IL-17 in the supernatant.

RESULTSTim-3 mRNA expression in PBMCs was significantly increased in asthmatic mice. The mRNA and protein expression of Tim-3 decreased significantly in the shRNA group. Compared with the negative groups, Th1 cell levels increased and Th17 cell levels decreased significantly in the asthmatic groups after Tim-3 shRNA interference. In the Tim-3 shRNA interference groups concentrations of IFN-γ increased significantly while IL-17 decreased significantly.

CONCLUSIONSSpecific Tim-3 shRNA effectively silences the expression of Tim-3 and change in Tim-3 expression could affect T cell differentiation.

Animals ; Asthma ; immunology ; therapy ; Cell Differentiation ; Female ; Gene Silencing ; Hepatitis A Virus Cellular Receptor 2 ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; genetics ; Receptors, Virus ; antagonists & inhibitors ; genetics ; Th1 Cells ; cytology ; Th17 Cells ; cytology

In order to determine the curative effect of multiglycosidorum Tripterygii and prednisone for treatment of children with chronic refractory idiopathic thrombocytopenic purpura (ITP), a total of 28 patients had been treated with multiglycosidorum Tripterygii and prednisone. Multiglycosidorum Tripterygii was given at a dose of 1 mg/kg body weight per day and the prednisone was given at a dose of 0.5 mg/kg body weight per day for two consecutive months. Bone marrow examination and blood platelet associated immunoglobin G (PAIgG) measurement were performed during the treatment. The results showed that in 21 patients acquired effectiveness, of whom, according the common criteria, 9 patients achieved curative effective, 7 achieved significant effectiveness and 5 effective. 7 patients did not achieve effectiveness. There were no serious side effects associated with therapy. The number of proplatelets on bone marrow smears was increased and the titers of IgG to platelet were restored to normal range in patients achieving remission. In conclusion, combination of multiglycosidorum triptergii with prednisone is effective and safe for treatment of chronic refractory idiopathic thrombocytopenic purpura in children.
Adolescent ; Child ; Child, Preschool ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Immunoglobulin G ; blood ; Immunosuppressive Agents ; administration & dosage ; Infant ; Male ; Platelet Count ; Prednisone ; administration & dosage ; Purpura, Thrombocytopenic, Idiopathic ; blood ; drug therapy ; Tripterygium

OBJECTIVETo examine the role of Polo-like kinase 1(PLK1) in the migration and invasiveness of human colorectal cancer cells.

METHODSNine colorectal cancer cell lines were cultured. Cell lines with the highest level of PLK1 expression was selected by PCR and Western blot. Three siRNA oligo segments targeting PLK1 were designed and selected cell lines transfected. Successful transfection was confirmed using real-time PCR and Western blot. Changes in migration and invasiveness of the selected cell line were evaluated by Transwell test.

RESULTSColorectal cancer cell line SW1116 was selected with the highest expression of PLK1 at both mRNA level and protein level. The expression of PLK1 in SW1116 was reduced by the three siRNA oligo segments to varying degrees, and the No.1 siRNA oligo segment was the most efficient. In migration test, the number of cells crossing through chambers in PLK1-siRNA group was 44 ± 14, which was lower than that in the negative control group (242 ± 40) and in blank control group(240 ± 38). In invasion test, the number of cells crossing through chambers in PLK1-siRNA group was 62 ± 3, which was lower than that in negative control group (207 ± 12) and in blank control group (211 ± 15). These differences were statistically significant(P<0.01).

CONCLUSIONPLK1 silencing by siRNA may inhibit the migration and invasiveness of colorectal cancer cells, suggesting that PLK1 might play an important role in the infiltration and metastasis of colorectal cancer.

Cell Cycle Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Neoplasm Invasiveness ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo explore the characteristics on molecular epidemiology of Mycobacterium tuberculosis isolates from hospitals in Tianjin area.

METHODSOne hundred M. tuberculosis isolated strains were collected in succession from August 16th-December 25th, 2005 in Tianjin Haihe Hospital and genotyped by spoligotyping and multiple locus variable number-tandem repeat(VNTR). Data was analyzed by cluster software. Based on the concept of Beijing lineage, it was determinate two sub-groups: atypical Beijing strains and W strain/typical family strains by multiplex and real-time PCR. The associations of subgroups with drug resistance and age were assessed by the chi2 test.

RESULTS96 M. tuberculosis strains were genotyped in which 91.7% (88/96) strains belonged to Beijing genotype (including 3 Beijing-like strains) by spoligtyping. VNTR typing could differentiate 60 genotypes among the 88 Beijing genotype strains. 93.2% of the Beijing lineage M. tuberculosis strains of this study belonged to W strain/typical Beijing family strains (82/88). No statistically significant differences were observed in the proportions of the two sub-groups in patients of different age, or drug resistance (P > 0.05).

CONCLUSIONThe M. tuberculosis Beijing genotype strains were dominated on tuberculosis hospital patients of Tianjin area. The discriminatory power of VNTR typing was higher than that of spoligtyping. The two sub-groups of Beijing lineage had been prevalent in Tianjin, however W strain/typical Beijing family strains were of preponderance.

Bacterial Typing Techniques ; China ; epidemiology ; Cluster Analysis ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Genotype ; Humans ; Minisatellite Repeats ; Molecular Epidemiology ; Mycobacterium tuberculosis ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Tuberculosis ; epidemiology

OBJECTIVETo explore the secure resection margin (RM) of hepatectomy for primary liver cancer (PLC) with the coexistence of cirrhosis or hepatitis by studying the correlations of the resected liver parenchyma volume with postoperative liver function, complication and RM clinically.

METHODSThe volume of tumor and the surrounding liver in resected liver specimen was measured and calculated in continuous 76 PLC patients prospectively, and the total liver parenchyma volume was measured and calculated using computed tomography (CT) images in former 40 patients. Under ideal circumstances, the surrounding liver volume, which would be resected theoretically, was calculated according to various sizes of tumors and RMs. The correlations of the resected liver volume or hepatic parenchyma-resected rate (HPRR) with postoperative liver function, complication and RM were analysed.

RESULTSThe RM was (5 +/- 7) mm in 76 patients. The volume of the tumors and the surrounding liver in the specimens were (107 +/- 203) cm(3) and (153 +/- 120) cm(3), respectively. In 40 patients, the total nontumorous liver volume using CT images was (1079 +/- 179) cm(3), and HPRR was (14 +/- 9)%. There were statistically significant differences in HPRR (P < 0.05) between three groups with complication score 0, 1-2 and 3-6 points, the value of the first group were lower than that of the third group at the level P < 0.05. The significant factors affecting liver function and complication are HPRR, the size of operation, the time of hepatic portal occlusion and the resected liver volume (P < 0.05) apart from preoperative liver function.

CONCLUSIONSWhen hepatectomy was performed in PLC patients with preoperative liver function of Child A grade and the coexistence of cirrhosis or hepatitis, 30% HPRR was a lower limit for greatly increasing the chance of developing serious postoperative complications, while 20% HPRR was a safe upper limit for achieves quick postoperative recovery or developing only a few mild complications. When PLC patients without macroscopic tumor thrombi or macrosatellites undergo hepatectomy, 10 mm RM is enough to ensure sufficient liver function residue and achieve complete micrometastasis clearance in liver parenchyma surrounding the lesion if the diameter of a tumor is less than 10 cm and 6 mm RM is enough to ensure sufficient liver function residue and obtain 99% micrometastasis clearance if the diameter of a tumor is greater than 10 cm, while with macroscopic tumor thrombi or macrosatellites, 20 mm RM is enough to ensure sufficient liver function residue and achieve 99% micrometastasis clearance if the diameter of a tumor is less than 6 cm.

Adult ; Aged ; Female ; Hepatectomy ; methods ; Hepatitis ; complications ; Humans ; Liver ; pathology ; Liver Cirrhosis ; complications ; Liver Neoplasms ; complications ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Prospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo explore the molecular mechanism underlying the biological function of lncRNA PTENP1 in bladder cancer.

METHODSExpressions of PTENP1, PTEN and miR-17 were examined by quantitative reverse transcriptase PCR (qRT-PCR) in 12 bladder cancer tissues. The expression of PTEN was examined by Western blotting in bladder cancer cell lines T24 and 5637 overexpressing PTENP1. Luciferase reporter assay was performed to confirm the targeting of miR-17 to PTENP1 and PTEN. T24 and 5637 cell lines with stable overexpression of PTENP1 and mir-17 were used to investigate effect of PTNE and miR-17 on the function of PTENP1 in bladder cancer.

RESULTSThe expression of miR-17 was up-regulated and PTENP1 and PTEN were down-regulated in bladder cancer tissues, where a positive correlation was found between PTENP1 and PTEN expressions and a negative correlation between PTENP1 and miR-17 (P<0.05). Overexpression of PTENP1 in bladder cancer cell lines T24 and 5637 obviously enhanced the expression of PTEN protein. miR-17 was found to target both PTENP1 and PTEN and promote the growth of bladder cancer. miR-17 could partially restore the tumor-suppressing activity of PTENP1 in bladder cancer.

CONCLUSIONBy binding with miR-17, lncRNA PTENP1 functions as a PTEN competing endogenous RNA (ceRNA) to suppress the progression of bladder cancer.

BACKGROUND: Although more and more accurate and personalized detection and assistive tools are used in the field of orthopedics with the development of technology, the preoperative CT is still the first-line examination choice of many orthopedic surgeons. Therefore, to master the lumbar spine CT imaging performance is not only the basis of spine surgeons, but also the cornerstone of spinal nail surgery. OBJECTIVE: Through CT imaging technique, we measured the parameters of lumbar pedicle imaging, and understood its internal structure as the basis for the choice of screws, in order to improve the safety and accuracy of nail surgery. METHODS: Totally 36 cases without lumbar vertebrae-related disease undergoing lumbar vertebra and abdominal cavity CT scan were randomly selected, including 20 males and 16 females at the age of (43.3±12.3) years. PACS was used to reconstruct lumbar vertebrae, and the related imaging parameters were measured. RESULTS AND CONCLUSION: (1) There was no significant difference between the left and right sides of the pedicle for paired t-test (P >0.05). (2) Pedicle sagittal angle had downward trend, the level of change was small; L4increased relative to L3; L5level was minimum at the average of -1.3°. Pedicle transverse angle had gradually increased trend, and maximum at L5; the average was 25.4°. (3) The length of the pedicle channel increased first and then decreased in the lumbar segment; the longest average was 52.2 mm in the L2segment, the shortest at L5, with an average of 47.4 mm. (4) The pedicle height was slightly narrowed at L1-L3levels; the smallest was 15.4 mm (L3mean), and reached the maximum in the L5at 22.7 mm (L5mean). Cancellous bone height also showed the same trend, the minimum in the L3at 10.2 mm, the maximum in the L5at 17.9 mm. (5) Lumbar pedicle width showed an upward trend, L1minimum of 7.7 mm, L5maximum of 12.5 mm. Cancellous bone width also became a similar upward trend; the narrowest L1, averagely 4.9 mm; the maximum L5, averagely 8.5 mm. (6) Overall, the upper lumbar pedicle channel was slender, while the lower lumbar spine was slightly short. (7) The parameters of the pedicle can be used as the basis for selecting the appropriate screw. At the same time, lumbar pedicle morphology has a certain difference, and there is a certain law. Accurately grasping its structure and image characteristics of pedicle screw fixation is the basis of surgery. The preoperative imaging data collection and assessment, combined with the general situation of patients, and the development of personalized programming, will help in improving the safety and accuracy of screw placement.