TRPV1 participates in the protective effect of propolis on colonic tissue of ulcerative colitis / 中南大学学报(医学版)

OBJECTIVES@#Ulcerative colitis (UC) is a type of inflammatory bowel disease (IBD) mainly characterized by inflammation, ulceration and erosion of colonic mucosa and submucosa. Transient receptor potential vanilloid 1 (TRPV1) is an important mediator of visceral pain and inflammatory bowel disease. This study aims to investigate the protective effect of water soluble propolis (WSP) on UC colon inflammatory tissue and the role of TRPV1.@*METHODS@#Male SD rats were randomly divided into 6 groups (n=8): a normal control (NC) group, an ulcerative colitis model (UC) group, a low-WSP (L-WSP) group, a medium-WSP (M-WSP) group, a high-WSP (H-WSP) group, and a salazosulfapyridine (SASP) group. The rats in the NC group drank water freely, and the other groups drank 4% dextran sulfate sodium (DSS) solution freely for 7 d to replicate the ulcerative colitis model. Based on the successful replication of the UC, the L-WSP, M-WSP, and H-WSP groups were given 50, 100, and 200 mg/kg of water-soluble propolis by gavage for 7 d, and the SASP group was given 100 mg/kg of sulfasalazine by gavage for 7 d. The body weight of rats in each group was measured at the same time every day, the fecal traits and occult blood were observed to record the disease activity index (DAI). After intragastric administration, the animals were sacrificed after fasted 24 h. Serum and colonic tissue were collected, and the changes of MDA, IL-6 and TNF-α were detected. The pathological changes of colon tissues were observed by HE staining, and the expression of TRPV1 in colon tissues was observed by Western blotting, immunohistochemistry, and immunofluorescence.@*RESULTS@#The animals in each group that drank DSS freely showed symptoms such as weight loss, decreased appetite, depressed state, and hematochezia, indicating that the model was successfully established. Compared with the NC group, DAI scores of other groups were increased (all P<0.05). MDA, IL-6, TNF-α in serum and colon tissues of the UC group were increased compared with the NC group (all P<0.01), and they were decreased after WSP and SASP treatment (all P<0.01). The results of showed that the colon tissue structure was obviously broken and inflammatory infiltration in the UC group, while the H-WSP group and the SASP group significantly improved the colon tissue and alleviated inflammatory infiltration. The expression of TRPV1 in colon tissues in the UC group was increased compared with the NC group (all P<0.01), and it was decreased after WSP and SASP treatment.@*CONCLUSIONS@#WSP can alleviate the inflammatory state of ulcerative colitis induced by DSS, which might be related to the inhibition of inflammatory factors release, and down-regulation or desensitization of TRPV1.

Optimization of Processing Technology of Fried Radix Paeoniae by Box-Behnken Response Surface Method / 中国药房

OBJECTIVE： To optimize the processing technology of fried Radix Paeoniae， and to provide reference for quality control of the processed products. METHODS： The content of paeoniflorin in fried Radix Paeoniae was determined by HPLC. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of acetonitrile-0.05 mol/L potassium dihydrogen phosphate solution （15 ∶ 85， V/V） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， and detection wavelength was set at 230 nm. The sample size was 10 μL. The appearance character of fried Radix Peaoniae were investigated by appearance color， crosssection color， hardness and smell. Taking the comprehensive score of appearance character and paeoniflorin content as evaluation index， the dosage， stir-frying temperature and stir-frying time were investigated. According to the results of single factor test， Box-Behnken response surface methodology was used to optimize above 3 factors. The optimized processing technology was validated. RESULTS： The linear range of paeoniflorin were 0.02-4.15 mg/mL （r＝0.999 9）； precision， stablity， repeatability and sample recovery rate meet the requirements. The optimal technology of fried Radix Paeoniae included the dosage of 374.60 g， frying temperature of 101.61 ℃， frying time of 20 min. Under optimal technology， comprehensive score of fried Radix Paeoniae ranged 97.39-98.82 in 6 times of parallel verification tests （RSD＝0.54％）， which was close to predicted value 98.18. The color of fried Radix Paeoniae was slightly deeper than Radix Paeoniae， which was crisp and fragrant. CONCLUSIONS： The optimized processing technology of fried Radix Paeoniae is stable and feasible， and is suitable for the preparation of the processed products.

Correlation between FibroScan, APRI Detecting the degree of esophageal and gastric varices of liver cirrhosis / 中华肝脏病杂志

Objective@#To investigate the predictive value of transient elastography (FibroScan), aspartate aminotransferase-to-platelet ratio index (APRI) in the detection of esophagogastric varices in patients with liver cirrhosis. @*Methods@#236 patients with liver cirrhosis who met the criteria were selected. All patients underwent gastroscopy. According to the degree of esophagogastric varices, patients were divided into four groups: none, mild, moderate, and severe. The patient's liver stiffness (LSM) and aspartate aminotransferase- to-platelet ratio index (APRI) were measured within 3 days of gastroscopy. One-way analysis of variance was used to compare multi-group data. The ROC curves of LSM, APRI, LSM+APRI in patients with liver cirrhosis with esophageal varices were plotted and their area under the ROC curve (AUC) were compared. @*Results@#The area under the ROC curve of LSM, APRI, LSM + APRI in patients with mild esophagogastric varices were 0.856, 0.900, and 0.906, respectively; moderate esophagogastric varices were 0.857, 0.924, and 0.923 respectively; and severe esophagogastric varices were 0.801, 0.903, and 0.901, respectively. @*Conclusion@#APRI and LSM+APRI have better predictive value for patients with cirrhosis who have esophagogastric varices.

Factors influencing neuropathic pain in patients with spinal cord injury / 中华物理医学与康复杂志

Objective To investigate the neuropathic pain (NP) accompanying spinal cord injury (SCI),and to analyze the related factors to provide references for its prevention and treatment.Methods Seventy SCI patients with NP were selected using the DN4 scale.Their age,gender,occupation,education level,monthly income,injury position,marital status and other data were surveyed using a general questionnaire.Their NP situation was surveyed using a simplified McGill pain questionnaire (SF-MPQ).Results The patients' average visual analogous scale (VAS) score was 4.37.Their average pain rating index (PRI) according to the SF-MPQ was 8.23,with the PRI-sensory and PRI-emotional components 5.23 and 3.00 respectively.The average degree of present pain intensity was 1.86,between mild pain and discomfort,and discomfort was the description most commonly used.The most common pain descriptor was prickling pain,followed by burning pain and bulge pain.85.7％ of the patients felt that their pain had an adverse effect on their affective state,and exhaustion occurred more often than any other descriptive words.Univariant analysis showed that the degree of injury,education level,marital status,monthly income,family support and medication history were all factors correlated with NP perceptions.Multi-variate logistic regression analysis showed that being unmarried and severity of injury were independent protective factors against NP.No family support,no medication and low income were independent risk factors for NP.Conclusion The type of neuropathic pain varies in patients with spinal cord injury.The intensity of the pain is mostly at a medium level.The emotional state of most patients was affected.Neuropathic pain involves many factors.Being unmarried and severely injured are independent protective factors,while lack of family support,no medication,and having low income were independent risk factors.

Tea Consumption is Associated with Increased Risk of Kidney Stones in Northern Chinese: A Cross-sectional Study / 生物医学与环境科学（英文）

Kidney stones are a common urinary system condition that can progress to kidney disease. Previous studies on the association between tea consumption and kidney stones are inconsistent. A cross-sectional study to investigate the association between tea consumption and kidney stones was conducted from 2013 to 2014 and recruited 9,078 northern Chinese adults. A total of 8,807 participants were included in the final analysis. Participants' prevalence of kidney stones was 1.07%, 1.73%, and 2.25% based on their tea consumption frequency of never, occasionally, and often groups, respectively. Compared with the 'never' group, the odds ratios (95% confidence intervals) for the occurrence of kidney stones were 1.57 (1.00-2.46) and 1.65 (1.06-2.57) in the 'occasionally' and 'often' groups, respectively. After adjusting for sex, age, and other potential confounding factors, tea consumption still significantly increased the risk of kidney stones. Tea consumption is independently associated with an increased risk of kidney stones in the investigated population, suggesting that a decrease in the consumption of tea may be a preventive strategy for kidney stones.

Scanning the SRSF2 gene mutations in acute myeloid leukemia by using high-resolution melting analysis / 实用医学杂志

Objective To study the SRSF2 mutations in acute myeloid leukemia (AML) patients by using high-resolution melting analysis (HRMA). Methods PCR-HRMA analysis was performed to screen SRSF2 mutations in 140 cases with AML, and the direct DNA sequencing was used to confirm the HRMA results. Results Five percent (7/140) of AML patients were found with heterozygous SRSF2 mutations, including one case of P95R mutation, two case of P95L mutation, and four cases of P95H mutation, the above mutations were confirmed by direct DNA sequencing. The maximal sensitivity of HRMA in detecting SRSF2 mutation was close to 10%. There were no difference in gender, age and blood parameters among cases with or without SRSF2 mutations (P > 0.05). The overall survival (OS) of patients with SRSF2 mutations was inferior to those without SRSF2 mutations in AML patients (P=0.016). Conclusions HRMA analysis was a convenient, rapid, specific, high-throughput technique for scanning of SRSF2 gene mutations in AML patients. SRSF2 mutation may predict the adverse prognosis in AML patients.

An enriched environment enhances synaptic plasticity and cognition post-stroke / 中华物理医学与康复杂志

Objective To observe the effects of an enriched environment (EE) on cognitive functioning and the synaptic plasticity of mice modeling post-stroke cognitive impairment (PSCI) and explore the possible mechanisms involved.Methods Mice modeling PSCI and sham-operated mice were randomly divided into 3 groups:sham-operated mice in a standard environment (the Sham+SE group),PSCI mice in a standard environment (the PSCI+SE group) and PSCI mice in an enriched environment (the PSCI+EE group).The cognitive functioning of all of the mice was quantified using a Morris water maze and their hippocampal long-term potentiation (LTP) was recorded using an electrophysiological method.The level of synaptophysin was detected using Western blotting.Synaptic ultrastructure in the hippocampus was imaged using electron microscopy.Results Compared with the Sham +SE group,the PSCI+SE group showed significantly poorer water maze performance and failed induction of contralateral LTP.Their average level of synaptophysin was significantly lower,and significant adverse changes in the synaptic ultrastructure of the hippocampus were observed,including a decreased number of synapses.The average width of the synaptic cleft,postsynaptic density and the interface curvature of the synapses were all less desirable.All of the measurements of the PSCI+EE group improved significantly compared to those of the PSCI+SE group,but were still significantly poorer than those of the Sham+SE group.Conclusions An enhanced environment can improve the cognitive functioning of mice modelling PSCI.It may be that an EE can improve synaptic plasticity in the hippocampus contralateral to the stroke.

Expression of CD44+/C-myc+cancer stem cells and its relationship with the prognosis of patients in colorectal tumors / 中国组织工程研究

BACKGROUND:Tumor recurrence results from the incomplete removal of cancer stem cel s with self-renewal characteristics, and then how to label and eliminate cancer stem cel s becomes the key to cancer treatment. OBJECTIVE:To observe the morphology,distribution and number of CD44+/C-myc+METHODS:Pathological tissues from 150 patients with colorectal cancer were taken to prepare tissue microarray, in order to observe and count CD44 cel s in colorectal cancer, and to explore the relationship between the expression and postoperative metastasis.+/C-myc+cel s by using immunohistochemical double staining. Patients were fol owed up through mobile phones , letters , and so on, and the relationship between the expression of CD44+/C-myc+cel s and postoperative metastasis were statistical y recorded.RESULTS AND CONCLUSION:There was no CD44+/C-myc+cel s in normal tissue and little in adenoma. A smal number of CD44+/C-myc+cel s distributed as dots or focal lesions in adenocarcinoma. The number of CD44+/C-myc+cel s was related to the degree of adenocarcinoma differentiation, depth of invasion, and lymph node metastasis (P<0.05). The univariate analysis showed that the overal survival rate and progression-free survival rate were associated with the number of CD44+/C-myc+cel s, lymph node metastasis and Ducks staging in adenocarcinoma (P<0.05). The multivariate analysis showed that the overal survival rate was related to CD44+/C-myc+cel amount and the progression-free survival rate was related to CD44+/C-myc+cel amount and the Ducks staging. Therefore, CD44+/C-myc+cel s are probably cancer stem cel s, and Ducks staging and CD44+/C-myc+cel amount are important prognostic factors for colorectal cancer.

Methylation situation of let-7a-3 in chronic myeloid leukemia and its clinical significance / 重庆医学

Objective To investigate the methylation situation of let‐7a‐3 promoter in patients with chronic myeloid leukemia (CML) and its clinical significance .Methods The methylation level of let‐7a‐3 promoter in the bone marrow mononuclear cells of 52 CML patients and 25 controls was detected by using the real‐time quantitative methylation‐specific PCR (RQ‐PCR) .Results The non-hypomethylation of let‐7a‐3 promoter was positive in 31 cases(59 .6% ) of 52 CML patients ,while only 1 case(4% ,1/25) in the control group ,the difference between the two groups were statistically significant (P0 .05) .The non-hypomethylation level of let‐7a‐3 in chronic phase and accel‐erate phase was significantly higher than that in blastic crisis of CML .Conclusion The hypomethylation level of let‐7a‐3 promoter is decreased with disease progression .

Variants of Interleukin-7/Interleukin-7 Receptor Alpha are Associated with Both Neuromyelitis Optica and Multiple Sclerosis Among Chinese Han Population in Southeastern China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Neuromyelitis optica (NMO) and multiple sclerosis (MS) are autoimmune demyelinating diseases of the central nerve system. Interleukin-7 (IL-7) and interleukin-7 receptor alpha (IL-7Rα) were proved to be important in the pathogenesis of both diseases because of the roles they played in the differentiations of autoimmune lymphocytes. The variants of both genes had been identified to be associated with MS susceptibility in Caucasian, Japanese and Korean populations. However, the association of these variants with NMO and MS has not been well studied in Chinese Southeastern Han population. Here, we aimed to evaluate the association of six IL-7 variants (rs1520333, rs1545298, rs4739140, rs6993386, rs7816065, and rs2887502) and one variant of IL-7RA (rs6897932) with NMO and MS among Chinese Han population in southeastern China.</p><p><b>METHODS</b>Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MassARRAY system) and Sanger sequencing were used to determine the variants of IL-7 and IL-7RA in 167 NMO patients, 159 MS patients and 479 healthy controls among Chinese Han population in southeastern China. Samples were excluded if the genotyping success rate <90%.</p><p><b>RESULTS</b>Statistical differences were observed in the genotypes of IL-7 rs1520333 in MS patients and IL-7RA rs6897932 in NMO patients, compared with healthy controls (P = 0.035 and 0.034, respectively). There was a statistically significant difference in the genotypes of IL-7 rs2887502 between MS and NMO patients (P = 0.014). And there were statistically significant differences in the rs6897932 genotypes (P = 0.004) and alleles (P = 0.042) between NMO-IgG positive patients and healthy controls.</p><p><b>CONCLUSIONS</b>The study suggested that among Chinese Han population in southeastern China, the variant of IL-7RA (rs6897932) was associated with NMO especially NMO-IgG positive patients while the variant of IL-7 (rs1520333) with MS patients. And the genotypic differences of IL-7 rs2887502 between MS and NMO indicated the different genetic backgrounds of these two diseases.</p>

Expression of SALL4 gene in patients with acute and chronic myeloid leukemia / 中国实验血液学杂志

The aim of this study was to investigate the expression level of the SALL4 gene and its clinical significance in patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). Real-time quantitative PCR (RQ-PCR) was performed to detect the expression level of SALL4 mRNA in bone marrow mononuclear cells (BMMNC) from 35 AML, 12 CML patients and 24 iron deficiency anemia patients as controls. The results indicated that the expression level of SALL4 in AML (0%-14%, median 1.43%) was obviously higher than that in controls (0% - 1%, median 0%) (P < 0.001). SALL4 expression was positive in 65.7% (23/35) AML patients. The frequency of SALL4 expression was in M2 (86.7%, 13/15) > M3 (75.0%, 6/8) > M1 (60.0%, 3/5) > M4 (14.3%, 1/7), and the difference among 4 groups was statistically significant (P = 0.008); there was no correlation of the frequency of SALL4 expression with the age, sex, white blood cell WBC count, hemoglobin concentration, platelet count and chromosomal abnormalities of AML patients (P > 0.05). All the 13 CML cases showed positive expression of SALL4 gene (1% - 128%, median 19.39%), which was higher than that in controls (P < 0.001). The analysis of receiver operating characteristic (ROC) curve showed the area under ROC curve (AUC) of AML and CML were 0.983 (95% confidence interval: 0.95 - 1.017) and 0.997 (95% confidence interval: 0.986 - 1.007) respectively. It is concluded that SALL4 expression is a common molecular event and can be considered as a molecular marker for assisting diagnosis of AML and CML.

Scanning of c-kit gene mutations in acute myeloid leukemias using high-resolution melting analysis / 中华血液学杂志

<p><b>OBJECTIVE</b>To detect the common mutations (D816V and N822K) of c-kit gene in acute myeloid leukemia (AML) using high-resolution melting analysis (HRM).</p><p><b>METHODS</b>HRM analysis was established to screen c-kit mutations in PCR products of c-kit exon 17 in 21 AML patients with t(8;21). PCR products were sequenced to confirm the mutation.</p><p><b>RESULTS</b>HRM analysis identified an aberrant melting curve in 6 cases (28.6%), which were confirmed by direct DNA sequencing as one D816V mutation and five N822K mutation.</p><p><b>CONCLUSION</b>HRM analysis is a convenient, rapid, specific and high-throughput technique for scanning c-kit gene mutation in AML.</p>

Quantification of prame gene transcript in chronic myeloid leukemia / 中国实验血液学杂志

This study was purposed to analyze the expression level of preferentially expressed antigen of melanoma (prame) transcript in the patients with chronic myeloid leukemia (CML) and explore its clinical significance. Real-time quantitative PCR (RQ-PCR) assay was used to detect the level of prame gene transcript in the bone marrow samples from 30 patients with CML and 15 patients with iron deficiency anemia (IDA). The results showed that CML patients had significantly higher prame mRNA level (0% - 772.25%, median 8.28%) than IDA cases (0% - 1.46%, median 0.19%) (p < 0.001). The level of prame gene transcript was significantly correlated with that of bcr-abl fusion gene transcript (r = 0.708, p < 0.001) in CML patients. Furthermore, 6 patients in blastic crisis (BC) and accelerated phase (AP) had significantly higher prame gene transcript than that of 24 cases in chronic phase (CP) (p = 0.007). In 2 CML patients with sequential samples, prame gene transcript increased in AP and BC, compared with in CP, and was consistent with the altering tendency of bcr-abl transcript. It is concluded that the level of prame gene transcript increases in CML which associates with the progression of the disease, prame gene transcript level can be used for monitoring the disease state.

Detection of BK virus infection in renal transplant recipients and clinical application / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application.</p><p><b>METHODS</b>132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKV-DNA to demonstrate the BK virus replication.</p><p><b>RESULT</b>Among 132 cases of renal transplant recipients, urinary decoy cell was found in 37 (28.0%) patients and the median time was 12 months after surgery. 32 (24.2%) patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16 (12.1%) recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy.</p><p><b>CONCLUSION</b>To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.</p>

Alteration of methylation status of death-associated protein kinase (dapk) gene promoter in patients with acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to analyze the methylation status of death-associated protein kinase (dapk) gene promoter in Chinese patients with acute myeloid leukemia (AML) and its relationship with clinical features. The methylation-specific PCR (MSP) technique was used to detect dapk promoter methylation in bone marrow samples from 112 cases of AML. The results indicated that gene dapk promoter hypermethylation was detected in 82 cases (73.2%), but not in 13 control group. There was no correlation of dapk gene hypermethylation with sex, age, WBC counts, platelet counts, hematologic parameters, chromosomal abnormalities and different subtypes of AML patients. It is concluded that dapk gene hypermethylation may be a common molecular event in AML.

Quantification of the PRAME transcripts in patients with acute myeloid leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the expression level and clinical significance of the preferentially expressed antigen of melanoma (PRAME) transcripts in patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>Real-time quantitative polymerase chain reaction with EvaGreen dye was established to detect the expression level of PRAME transcripts in the bone marrow mononuclear cells of 56 AML cases and 20 controls. The clinical association of PRAME transcripts was analyzed.</p><p><b>RESULTS</b>The PRAME transcripts were 0-1.46% (median 0.18%) and 0-21 618.09% (median 9.79%) in controls and AML cases, respectively (P< 0.01). Among the FAB subtypes, those with M1, M2, M3 and M4 had significantly higher level of PRAME transcripts than controls. However, those with M5 had similar level of PRAME transcripts as controls. There was a significantly negative correlation between the PRAME transcripts and cytogenetic risk groups (r= -0.438, P= 0.001). Cases in low risk had significantly higher level of PRAME transcripts than those in intermediate and high risk. Among cases with AML-M2, those with t(8;21) had significantly higher level of PRAME transcripts (135.06% -21 618.09%, median 2201.88%) than those without t(8;21)(0.14% -1696.30%, median 17.97%)(P= 0.002). In a patient with sequential samples, PRAME transcripts significantly decreased after induction therapy and significantly increased after relapse.</p><p><b>CONCLUSION</b>The PRAME transcript was highly expressed in AML patients and was a favorable marker of prognosis. Quantification of PRAME transcript can be used in monitoring disease status of AML.</p>

Quantification of GRAF gene expression in patients with acute myeloid leukemia using EvaGreen real time quantitative PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To quantify the expression level of GRAF gene in acute myeloid leukemia (AML) and analyze its clinical significance.</p><p><b>METHODS</b>The EvaGreen real-time quantitative polymerase chain reaction (RQ-PCR) assay was established and performed to measure the GRAF gene transcripts in 71 cases with AML and 21 with nonmalignant hematological diseases. The clinical correlation of GRAF expression was analyzed.</p><p><b>RESULTS</b>The established EvaGreen RQ-PCR assay had good specificity, reproducibility and sensitivity. The GRAF expression level was significantly lower in AML (0.01%-169.75%, median 3.82%) than that in controls (14.49%-126.85%, median 56.04%) (P<0.05). There was no correlation between the level of GRAF transcript and the sex, age, hematologic parameters, FAB subtypes and karyotypic groups (P>0.05).</p><p><b>CONCLUSION</b>The GRAF gene was down-regulated in AML, which might play a role in the leukemogenesis.</p>

Morphology and distribution of EpCAMhigh/CD44+ colorectal cancer stem cells / 中国组织工程研究

BACKGROUND: Cancer stem cells have bean a hot topic in life science research. EpCAMhigh/CD44+ colorectal cancer stem cells are beneficial to observe morphological characteristics and distribution of cancer stem cells.OBJECTIVE: To investigate the quantity, location, distribution and hematoxylin-easin staining morphologic features of colorectal cancer stem cells (Co-CSCs).DESIGN, "rIME AND SETrlNG: The observational study was performed at the Department of Pathology, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA from March to August 2008.MATERIALS: A total of 67 colorectal cancer paraffin embedding samples were collected from 200512007 archive at the Department of Pathology, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA, comprising 27 males and 40 females, aged 29 90 years.METHODS: EpCAM and CD44 was used to label cancer stem cells in paraffin embedding colorectal cancer samples. The locations of EpCAM and CD44 double positive cells were detected by SP(streptavidin HRP) immunohistochemical staining and double immunohistochemical staining. The morphologic features of them were investigated on hematoxylin-easin staining at the same position.MAIN OUTCOME MEASURES: Number, location, distribution of EpCAM and CD44-posltive cells, and morphology following hematoxylin-eosin staining.RESULTS: The number of double-positive cells accounted for 0.1% to 30.0% of all tumor cells, and the cells were scattered or distributed focally along the basement of glandular-like structure. The cells with scarcely cytoplasm were cube or oval, and its nucleus was oval or high cylindrical, deep stained and homogeneous; The quantity of double-positive cells were negatively correlated with the differentiation of colorectal cancer.CONCLUSION: Cancer stem cells are the causes of tumor development, metastasis and recurrence and drug resistance. To observe the number, location, distribution and morphology of Co-CSCs and to analyze the relationship between Co-CSCs and pathological parameters will provide guidance for the diagnosis, staging, prognostic evaluation and clinical prognosis of cotorectal carcinoma.

Quantification of PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia by using real-time quantitative PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish and evaluate a real time quantitative PCR (RQ-PCR) method for detection and quantification the PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Three pairs of primers and TaqMan probe were designed for detecting the most frequent PML/RAR alpha transcripts (L-form, S-form and V-form) and normal ABL was used as an internal control. A real time PCR condition was established to detect PML/RAR alpha and ABL positive templates with a series of dilutions. To evaluate this assay, bone marrow samples from 6 APL patients were detected.</p><p><b>RESULTS</b>In repeated tests, maximal sensitivities of 10 copies/microL were obtained, while reproducible maximal sensitivity achieved 100 copies/microL. In 10 normal controls, no amplified fluorescent signals were detected. The median absolute and normalized amount of PML/RAR alpha fusion gene transcripts were 4.27 x 10(3)-3.36 x 10(5) copies/50 ng (median 4.33 x 10(4)copies/50 ng) and 29.38%-600.53% (median 48.12%) respectively. One case showed significant decrease of PML/RAR alpha fusion gene transcripts after induction therapy compared to that at the time of diagnosis, while the fusion transcripts significantly increased after relapsed.</p><p><b>CONCLUSION</b>RQ-PCR is a sensitive, reliable quantitative assay and can be used in the diagnosis of APL and measurement of MRD.</p>

Expression of PML-RAR alpha fusion transcripts detection by using RQ-PCR / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish the real time quantitative PCR (RQ-PCR) assay according to 'Europe Against Cancer' (EAC) program and analyze the results of detection and quantification of different PML-RAR alpha transcript isoforms in patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Three RQ-PCR systems were performed to detect the most frequent PML-RAR alpha transcripts (L-form, S-form and V-form) in 30 APL patients and the RQ-PCR end-point products were identified by electrophoresis.</p><p><b>RESULTS</b>S-form RQ-PCR system could amplify positive signals of three isoforms in all of 30 cases, and V-form RQ-PCR system could do so in both L-form and V-form positive cases, however, L-form RQ-PCR system could only do so in L-form-positive cases. Electrophoresis and sequencing of end-point products amplified by S-form RQ-PCR system revealed three bands in each of L-form (621 bp, 477 bp and 218 bp) and V-form (567 bp, 423 bp and 218 bp) positive patients samples.</p><p><b>CONCLUSIONS</b>RQ-PCR, sensitive and reliable, can be used for monitoring the minimal residual disease in APL patients, however, its results should be interpreted carefully if it is used for detection of PML-RAR alpha fusion transcripts prospectively.</p>