Paraffin Embedding ; methods ; Tissue Array Analysis ; instrumentation ; methods

Antigens, CD34 ; metabolism ; Female ; Humans ; Hypoglycemia ; etiology ; Immunohistochemistry ; Middle Aged ; Pleura ; metabolism ; pathology ; surgery ; Solitary Fibrous Tumor, Pleural ; complications ; metabolism ; surgery ; Treatment Outcome ; Vimentin ; metabolism

Objective To study the feasibility and clinical values of diffusion tensor tractography (DTT)in the spinal cord at 3.0 T MR.Methods Forty patients with spinal cord compression including cervical cord herniation and cervical spondylosis(30 cases),tumors in spinal canal(9 cases)and old injury in cervical vertebrae(1 cases)and 20 healthy volunteers participated in this study.Single-shot spin- echo echo-planar diffusion tensor sequence for tractography of the spinal cord was performed.The fibers of spinal cord were visualized by using fiber tracking software.Results On the DTT maps,the normal spinal cord was depicted as a fiber tract showing color-encoded cephaloeaudally,which indicated anisotropy in the cephalocaudal direction.By setting two ROI,the main spinal cord fiber tracts,such as corticospinal or spinothalamic tract,were visualized.The tracts from two sides of the brain did not completely cross.It was asymmetric in the number of tracts on the two sides in most normal subjects(8/10).The tracts of all patients with cord compression were seen oppressed or damaged in different degrees.The DrrT in patients with cervical spondylosis and extramedullary-intradural neurolemmoma demonstrated that tracts were oppressed but not damaged.The DTT in one ependymoma showed that tract was markedly compressed and slightly damaged.Conclusion DTT is a promising tool for demonstrating the spinal cord tracts and abnormalities,can provide useful information for the localization of compression and evaluation of the impairment extent on the white matter tracts of the spinal cord.

OBJECTIVETo explore the efficacy on knee osteoarthritis (KOA) treated with different acupuncture methods at different stages.

METHODSOne hundred and eighty cases of KOA were divided into stagnation stage, fascia cramp stage and tendon lesion stage according to the condition of disease, 60 cases in each stage. Each stage was randomized into a staging treatment group and an electroacupuncture (EA) group, 30 cases in each one. In the staging treatment group, acupuncture at the tendon points of meridians and electric thermal needling method were adopted for the cases at the stagnation stage; the small needle-knife therapy and bleeding method were used for the cases at the fascia cramp stage; and the electric thermal therapy with thick silver needles was applied for the cases at the tendon lesion stage. In the EA group, EA was applied for the cases of all the three stages at Liangqiu (ST 34), Xuehai (SP 10), Yanglingquan (GB 34), etc.

RESULTSThe total effective rate was 96.1% (87/90) in the staging treatment group and was 91.1% (82/90) in the EA group, without significant difference in comparison (P > 0.05). The controlled and remarkably effective rate (88.9%, 80/90) in the staging treatment group was higher apparently than (62.2%, 56/90) in the EA group (P < 0.001). And in the EA group, with the disease progression and the further disease stages, the controlled and remarkably effective rate was reduced. After treatment, the scores of symptoms and signs were decreased in both groups (both P < 0.001), the improvement in the staging treatment group was superior to that in the EA group (P < 0.001).

CONCLUSIONThe different acupuncture methods at the three stages improve obviously the clinical effect and are highly targeted. The mechanism of the three stages on "meridian muscle region pathology" and the treatment based on the disease stages can be the effective approach to KOA.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; pathology ; therapy

OBJECTIVETo measure the expression of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (PKB) in liver tissue among low-birth-weight newborn rats treated with L-arginine (L-Arg) in early life, and to investigate the effect of L-Arg on insulin resistance.

METHODSEighteen pregnant rats were randomly divided into three groups: control, model and intervention (n=6 each). The control group was fed with normal protein feed (protein content=21%) during pregnancy to establish a normal-birth-weight newborn rat model, and the model and intervention groups were fed with low-protein feed (protein content=10%) during pregnancy to establish a low-birth-weight newborn rat model. Newborn rats from the three pregnant rat groups were also assigned to control, model and intervention groups. During 21 days of lactation, maternal rats in the control and model groups were fed with normal protein feed and normal drinking water, while maternal rats in the intervention group were fed with normal protein feed and drinking water rich in L-Arg (200 mg/kg·d). After ablactation, the three groups of newborn rats were fed with normal protein feed and normal drinking water. Liver tissue samples were collected from these newborn rats at 1, 3 and 8 weeks after birth. Protein expression of PI3K and PKB in liver tissue was measured by Western blot.

RESULTSAt 1 week after birth, the newborn rats in the intervention group had significantly higher protein expression of PI3K than in the model group (P=0.045), but there was no significant difference when compared with the control group (P=0.503). At 8 weeks after birth, the newborn rats in the intervention group had significantly higher protein expression of PKB than the model group (P=0.039), but there was no significant difference when compared with the control group (P>0.05).

CONCLUSIONSA supplement of L-Arg in early life can boost protein synthesis, increase protein expression of PI3K and PKB in liver tissue, promote insulin signaling and reduce insulin resistance.

Animals ; Animals, Newborn ; Arginine ; pharmacology ; Birth Weight ; Female ; Liver ; metabolism ; Male ; Phosphatidylinositol 3-Kinases ; genetics ; Phosphorylation ; Pregnancy ; Proto-Oncogene Proteins c-akt ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the regional spread of micrometastatic nodules in the mesorectum from low rectal cancer, and provide further pathological evidence to optimize radical resection procedure for rectal cancer.

METHODSA total of 62 patients with low rectal cancer underwent low anterior resection and total mesorectal excision (TME) was included in this study. Surgical specimens were sliced transversely and serial embedded blocks were made at 2.5 mm interval, and paraffin sections were stained with hematoxylin and eosin. The mesorectum on whole-mount sections was divided into three regions: outer region of mesorectum (ORM), middle region of mesorectum (MRM) and inner region of mesorectum (IRM). Microscopic spread were examined microscopically on the sections for the distribution in different mesorectal regions, frequency, types, involvement of lymphatic system and correlation with the primary tumor.

RESULTSMicroscopic spread of the tumor in mesorectum and ORM was observed in 38.7% (24/62) and 25.8% (16/62) of the patients, respectively. Circumferential resection margin (CRM) involved by microscopic tumor foci occurred in 6.5% (4/62) of the patients, and distal mesorectum (DMR) involvement was recorded in 6.5% (4/62) with a spread extent within 3 cm of distal border of the main lesions. Most (20/24) of the patients with microscopic spread in mesorectum were in TNM stage III.

CONCLUSIONResults of the present study support that complete excision of mesorectum without destruction of the ORM is essential for surgical management of low rectal cancer, and an optimal DMR clearance resection margin should not be less than 4 cm.

Adenocarcinoma ; pathology ; surgery ; Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mesentery ; pathology ; surgery ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Neoplasm, Residual ; Neoplastic Cells, Circulating ; pathology ; Peritoneal Neoplasms ; pathology ; surgery ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; surgery

OBJECTIVETo investigate the expression of phosphatase of regenerating liver-3 (PRL- 3) mRNA and evaluate its relationship with tumor invasion and metastasis in human colorectal carcinoma.

METHODSThe expression level of PRL-3 mRNA was examined semi-quantitatively in surgically resected tumor specimens, paired paratumor normal tissues from 46 CRC patients, metastatic lymph nodes and liver metastases from 18 cases with metastasis,adenoma tissues from 6 patients with colorectal adenoma (CRA). In addition,the mutation of PRL-3 gene was examined by PCR-SSCP.

RESULTSThe PRL-3 mRNA level was increased in primary CRC tissues as compared with paired paratumor normal tissues (1.6+/- 0.7 vs. 0.4+/- 0.1, P< 0.01), while no significant difference of its expression was found between CRA tissues and their adjacent normal mucosae (P> 0.05). However,the PRL-3 mRNA levels of liver metastases (2.1+/- 0.8) in 12 cases and metastatic lymph nodes (3.3+/- 1.0) in 6 cases were significantly higher compared with the matched primary lesions, normal tissues and negative-lymph nodes (P< 0.01). There was significant relation of the expression of PRL-3 mRNA with the clinicopathological features including Dukes stage, invasion depth and metastasis (P< 0.05), but no relation with sex,tumor size,degree of differentiation was found (P> 0.05). Abnormal electrolysis band was found in 1 of 6 cases with liver metastasis by PCR-SSCP analysis.

CONCLUSIONPRL-3 gene plays an important role in tumor invasion and metastasis and may associated with carcinogenesis and development of CRC. There might exist some unknown mechanisms of overexpression and mutation of PRL-3 gene in CRC.

Adult ; Aged ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Gene Expression ; Humans ; Liver Neoplasms ; secondary ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Proteins ; metabolism ; Protein Tyrosine Phosphatases ; metabolism ; RNA, Messenger ; metabolism

Objective:Effect of gut microbiota on human health has become a hotspot in recent years with the emergence of new microbial technology.B.bifidum is a traditional probiotics and has been widely used in food and medicine.But the metabolism and function of B.bifidum ATCC 29521 has not been well documented.To investigate colonization and growth of B.bifidum ATCC 29521 in mice intestine,fecal B.bifidum concentrations were analyzed by real-time fluorescent quantitative PCR.Methods:Female C57BL/6 mice were orally gavaged with eigher a single dose of 1 × 109 CFU B.bifidum ATCC 29521 or continuous doses of 1 × 109/day for 3 weeks.Feces were collected at 0,2,4,6,8,10,12,16,20,24 h and at 10,7,14,21,24,28 d,respectively.Total DNA was isolated from the feces using the Qiagen DNA stool extraction kit according to the manufacturer's instructions.The linear standard curve was established by each dilution degrees of B.bifidum ATCC 29521 and their corresponding CT values.The curve equation was y =0.1835x + 9.0628 and R2 was 0.9994.The concentration of B.bifidum ATCC 29521 in mice feces were calculated by substituted CT values obtained by RT-PCR into the curve equation.Results:The curves of B.bifidum ATCC 29521 concentration rose at first and then reduced gradually.In single dose group,the concentration of B.bifidum ATCC 29521 began to increase at 4h after oral gavage,reaching its peak 6.0 × 107 CFU/g at 10h and then decreased gradually.The biggest drop occurred at the period between 12 h and 16 h after B.bifidum treatment.In successive administration group,the concentration rose at the highest rate in the first week when it achieved 2.0 × 107CFU/g and kept on inceasing to 1.0 × 108CFU/g in the next week.However,the concentration did not rose up significantly in the third week.It means that the concentration of B.bifidum ATCC 29521 in mice intestine reached platform in the second week after oral gavage.The concentration of B.bifidum ATCC 29521 significantly decreased at 24h and one week after B.bifidum treatment course in two group repectively and was still higher than baseline before oral gavage.Conclusion:Once the B.bifidum ATCC 29521 supplement was discontinued,the concentration fell down in short time.B.bifidum ATCC 29521 could not sustain colonization and growth in mice intestine without external supplement.It is necessary to provide daily supplemention for at least two weeks and to keep on in order to maintain sufficient concentration.

OBJECTIVETo investigate the mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test (IRP).

METHODSTwenty-one iron overloading, 26 non-iron overloading IRP patients and 10 normal controls were enrolled in this study. The expressions of ROS, Bcl-2, Caspase-3 and apoptosis of BMMNC were analyzed by flow cytometry (FCM). Antioxidants were added to iron overloading IRP BMMNC, and then the changes of indices above were detected by FCM. The number and apoptosis of T lymphocytes of IRP patients were also detected.

RESULTSROS and apoptosis of BMMNC, myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones and normal controls (P < 0.05). The expressions of Bcl-2 on BMMNC, erythrocytes and stem cells of iron overloading IRP patients were significantly lower than those of non-iron overloading IRP ones (P < 0.05). The levels of Caspase-3 on myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones and normal controls (P < 0.05). After treatment with antioxidants, the expressions of ROS, Caspase-3 and apoptosis of iron overloading IRP BMMNC significantly decreased, but opposite for Bcl-2. The percentages of CD4(+) lymphocytes [ ( 40.86 ± 8.74）%] and CD4(+)/CD8(+) (1.44 ± 0.36) in PB of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones [(35.96 ± 7.03)% and 1.14 ± 0.37] and normal controls [(28.00 ± 6.73)% and 0.79 ± 0.21], respectively (P < 0.05), as opposite for CD8(+) lymphocytes (P < 0.05). The apoptosis of CD8(+) lymphocytes [(27.35 ± 10.76)%] and the ratio of CD8(+) apoptosis/CD4(+) apoptosis (2.51 ± 0.81) in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones [(15.47 ± 8.99)%] and normal controls (1.39 ± 0.47), respectively (P < 0.05). The apoptosis of erythrocytes and stem cells coated with auto-antibodies in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP and normal controls.

CONCLUSIONMechanisms underlying bone marrow damage by iron overload might be through the follows: ①The increased ROS induced by excessive iron deposition affected the expressions of Caspase-3 and Bcl-2, which caused more BMMNC apoptosis; ②The abnormal number and ratio of T lymphocytes caused by iron overload aggravated the abnormality of immunity of IRP; ③Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.

Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Case-Control Studies ; Caspase 3 ; metabolism ; Coombs Test ; Female ; Humans ; Iron Overload ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; physiopathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Young Adult

The aim of this study was to investigate the effect of suppression of nicotinamide phosphoribosyltransferase (NAMPT) expression on imatinib-sensitivity in chronic myelogenous leukemia (CML) cell line K562 and its mechanisms, NAMPT siRNA was synthesized and transfected into K562 cells. PI/Calcein staining technique was used to determine survival rate of transfected K562 cells at 48th hour after exposure to 1 µmol/L imatinib. MTS method was used to determine the proliferation changes of transfected K562 cell at 48th hour after exposure to different doses of imatinib, then half inhibitory concentration (IC(50)) was calculated. Expression of NAMPT at 3rd-48th hour after exposure to 1 µmol/L imatinib was determined by Western blot. To explore the effect of NAMPT-siRNA and imatinib on the expression of apoptosis-related genes, the microarray data from NCBI GEO Data-Sets was analyzed, then the results were confirmed by Western blot. The luciferase reporter assay was used to determine the effect of NAMPT and imatinib on transcriptional activity of NF-κB transcription factors. The results showed that after exposure to 1 µmol/L imatinib for 3 - 48 h, there was no significant change of NAMPT expression in K562 cells. The expression of NAMPT could be effectively inhibited by the NAMPT-siRNA. After exposure to 1 µmol/L of imatinib for 48 h, the survival rate of NAMPT-siRNA interference group was lower than that of negative control group (P < 0.05), indicating that suppression of NAMPT expression can increase the sensitivity of K562 cells to imatinib and enhance the killing effect of imatinib on K562 cells. The IC(50) of imatinib in NAMPT-siRNA interference group was the lowest compared with that of control group (P < 0.05) after exposure to different concentrations of imatinib for 48 h, the fitted survival curves showed that the slope of NAMPT-siRNA interference group was the largest ranging between 0.01 - 0.1 µmol/L of imatinib. Data mining of expression profiling indicated that the anti-apoptotic factor Bcl-2 decreased in K562 cells treated with either NAMPT-siRNA or imatinib, which was confirmed by Western blot. The inhibitory effect was much more significant when both NAMPT-siRNA and imatinib were used. The results of luciferase reporter assay showed that either NAMPT-siRNA or imatinib decreased transcriptional activity of NF-κB. The decreased effect was much more significant when both NAMPT-siRNA and imatinib were used. It is concluded that survival of K562 cells affected by imatinib may not be due to regulation of expression of NAMPT. When expression of NAMPT decreases, the K562 cells are more sensitive to imatinib, this may be related with the decreased transcriptional activity of NF-κB and its downstream effector Bcl-2.
Benzamides ; Cytokines ; antagonists & inhibitors ; metabolism ; Fusion Proteins, bcr-abl ; metabolism ; Humans ; Imatinib Mesylate ; K562 Cells ; NF-kappa B ; metabolism ; Nicotinamide Phosphoribosyltransferase ; antagonists & inhibitors ; metabolism ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrimidines ; pharmacology