AIM:To determine the contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae and Semen Euphorbiae Pulveratun from different habitats.METHODS:A kromasil C18 column(150 mm?4.6 mm,5 ?m) was used with methanol-water(68 :32) as the mobile phase and UV detection was at 280 nm.RESULTS:The linear ranges of euphorbiasteroid and lathyrol diadetate benzozte were from 0.200 ?g to 1.200 ?g and 0.206 ?g to 1.651 ?g(r=0.999 7,r=0.9999).The contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae from different habitats ranged from 0.450 8% to 0.625 1% and 0.692 3% to 0.752 9%.The contents of two kinds of compositions in Semen Euphorbiae Pulveratun ranged from 0.156 8% to 0.221 1% and 0.232 2% to 0.283 0%.CONCLUSION:The method for quantitating euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae is accurate and reliable,and can be used to control the quality of toxicity Semen Euphorbiae and Semen Euphorbiae Pulveratun.

AIM:To determine the contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae and Semen Euphorbiae Pulveratun from different habitats.METHODS:A kromasil C_(18) column(150 mm×4.6 mm,5 μm)was used with methanol-water(68:32)as the mobile phase and UV detection was at 280 nm.RESULTS:The linear ranges of euphorbiasteroid and lathyrol diadetate benzozte were from 0.200 μg to 1.200 μg and 0.206 μg to 1.651 μg(r = 0.999 7,r = 0.999 9).The contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae from different habitats ranged from 0.450 8% to 0.625 1 % and 0.692 3% to 0.752 9%.The contents of two kinds of compositions in Semen Euphorbiae Pulveratun ranged from 0.156 8% to 0.221 1% and 0.232 2% to 0.283 0%.CONCLUSION:The method for quantitating euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae is accurate and reliable,and can be used to control the quality of toxicity Semen Euphorbiae and Semen Euphorbiae Pulveratun.

Objective To explore the role of basic leucine zipper ATF-like transcription factor (BATF) in active tuberculosis, and to provide clues for diagnosis and therapy of tuberculosis.Methods Sixteen patients with active tuberculosis (ATB), ten cases of latent tuberculosis infection (LTBI) and fourteen healthy controls (HC) were included in this study.Flow cytometry was applied to detect the expressions of BATF and programmed death-1 (PD-1) in T lymphocytes, and the changes of BATF by blockade of PD-1/PD-L pathway using specific blocking antibody antiPD-1, antiPD-L1 and antiPD-L2.The expressions of BATF were compared using Mann-Whitney U test.And the relation of BATF and PD-1 was analyzed using Pearson correlation analysis.Results The CD4+ T lymphocytes expressing BATF accounted for 5.16% (2.96%,8.71%) of CD4+ T lymphocytes in ATB group, which was higher than 1.05% (0.40%,1.27%) in LTBI group and 0.71%(0.43%,1.21%) in HC group, and the difference were statistically significant (U value were 6.5 and 9.0, respectively, both P<0.01).The CD8+ T lymphocytes expressing BATF accounted for 4.10% (2.27%,8.17%) of CD8+ T lymphocytes in ATB group, which was higher than 0.55% (0.34%,1.18%) in LTBI group and 0.84% (0.41%,1.29%) in HC group, and the difference were statistically significant (U value were 5.0 and 8.0, respectively, both P<0.01).Furthermore, the percentage of BATF+ PD-1+ CD4+ T lymphocytes in the peripheral blood of ATB was significantly higher than those in the peripheral blood of LTBI and HC, the difference were statistically significant (Uvalue were 16.0 and 14.5, respectively, both P<0.01), and the percentage of BATF+ PD-1+ CD8+ T lymphocytes in the peripheral blood of ATB was significantly higher than those in the peripheral blood of LTBI and HC, the difference were statistically significant (Uvalue were 10.0 and 16.5, respectively, both P<0.01).In addition, there was a positive correlation between the percentage of BATF+ T cells and PD-1+ T cells, both in CD4+ T cells (r=0.676,P=0.016) and CD8+ T cells (r=0.610,P=0.035).The expressions of BATF both in CD4+ T cells and CD8+ T cells were decreased followed by blockade of PD-1/PD-L pathway (P<0.05).Conclusions BATF may be involved in the regulation of immune pathogenesis of tuberculosis.In order to provide a theory for anti-tuberculosis immunotherapy fargeting BATF, further research need to be proceeded.

Objective To compare the MRI manifestations and characteristics of ≤ 3 cm nonepithelial hepatic angiomyolipomas (HAML)and small hepatocellular carcinoma (SHCC),then improve the preoperative diagnostic accuracy.Methods A retrospective analysis of 20 patients ≤3 cm nonepithelial HAML and 26 cases of SHCC,confirmed by clinical pathology,with both in clinical data and MRI characteristics.Results ≤3 cm nonepithelial HAML commonly occurs in women;The enhancement patterns“wash in and wash out”in 1 1 cases,6 cases “wash in but slow out”,3 cases with delayed enhancement;Mature adipose tissue found in 5 cases, 3 cases has pseudocapsule enhancemen in delayed phase,13 cases can see central vessels;While SHCC often occurs in men,with“wash in and wash out”enhancement pattern in 23 cases,1 7 cases with pseudocapsule enhancemen in delayed phase,12 cases can see central vessels in lesions.ADC values for SHCC was significantly lower than that for ≤3 cm nonepithelial HAML,the ADC values of SHCC significantly lower than the surrounding liver parenchyma;The ADC values between ≤3 cm nonepithelial and liver parenchyma around has no significant differences.ADC values of liver parenchyma with liver cirrhosis was lower than that without cirrhosis.Conclusion The existence of mature adipose tissue,the MRI enhancement pattern and the value of ADC can help to distinguish between ≤3 cm nonepithelial HAML and SHCC,then improve the preoperative diagnostic accuracy.

Objective To investigate the mechanism of hypoxia regulate osteopontin (OPN) secreting by mature dendritic cells (mDCs). Methods CD14 + cells were enriched using anti-CD14 immunomagnetic beads, for inducing to mDCs, CD14 + cells were cultured with GM-CSF and IL-4 in hypoxia or normoxiain vitro. Concentration of OPN and TGF-β1 in supernatant were detected by sandwich ELISA, OPN mRNA detected by RT-PCR. Approach regulating function of A2 R in expressing of OPN by mDCs by using NECA (surrogate of adenosine), A2R agonist (CGS21680), A2R antagonist (SCH58261) and investigate role of TGF-β1 in this process by using rhTGF-β1 and anti-TGF-β1 Ab. Results Hypoxia inreased the level of OPN and OPN mRNA in mDCs, and this effect could be reversed by A2 R antagonist. Under normoxia,both NECA and A2R agonist (CGS21680) could upregulate the level of OPN and OPN mRNA in mDCs significantly, but this positive effect could be reversed by A2 R antagonist. A2 R played a role in regulating TGF-β1, and confirmed TGF-β1 involved in regulation of OPN by using rhTGF-β1 and anti-TGF-β1 Ab. Conclusion High adenosine induce the generation of TGF-β1 through the A2R on mDCs, and then TGF-β1 raise the OPN secreting by mDCs.

OBJECTIVETo isolate the side population (SP) and non-side population (NSP) cells from A431 cells and compare their difference in tumorigenicity in mice and the expression profiles of epithelial-mesenchymal transition (EMT) markers.

METHODSA431 cells stained with Hoechst 33342 were sorted with flow cytometry. The isolated SP cells and NSP cells were inoculated into NOD/SCID mice and the tumorigenicity of the cells was observed. EMT markers E-cadherin, β-catenin, vimentin, AXL, and Erbb3 in the tumor tissues were detected by immunohisto-staining.

RESULTSThe tumors generated by SP cells were larger than those by NSP cells in NOD/SCID mice. Compared with the tumors generated by NSP cells, the cells in the periphery of tumors generated by SP cells showed up-regulated expressions of AXL, vimentin and β-catenin and down-regulated ERBB3 and E-cadherin.

CONCLUSIONThe SP cells in A431 cells have a strong tumorigenicity and show more EMT phenotypes in tissues.

Animals ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplastic Stem Cells ; metabolism ; Side-Population Cells ; metabolism ; Skin Neoplasms ; metabolism ; pathology

OBJECTIVETo assess the efficacy of full-thickness excision using transanal endoscopic microsurgery (TEM) in the treatment of rectal neuroendocrine tumors (NET).

METHODSClinicopathological and follow-up data of 90 rectal NET patients who underwent TEM between December 2006 and December 2016 at our department were retrospectively analyzed. TEM was performed as primary excision in 66 patients and as the second complete surgery because of suspected positive margin of samples after colonoscopic polypectomy in 24 patients.

RESULTSTEM was successfully performed in all the rectal NET patients, and in 10 patients(41.7%,10/24) among those undergoing the second excision, postoperative pathologic results showed remnant tumor. The mean diameter of all the tumors was (1.03±0.46) cm, and the mean tumor diameter of primary excision and secondary excision was (1.10±0.50) and (0.84±0.23) cm respectively (t=2.454, P=0.016). The mean distance from tumor low margin to anal verge was (7.7±1.8) cm for all the patients, and such distance for those undergoing primary excision and secondary excision was (7.4±1.7) cm and (8.4±1.8) cm respectively (t=2.233, P=0.028). Of all the patients, the mean intra-operative blood loss was (13.7±5.1) ml, and the mean operation time was (56.6±12.1) min. The intra-operative blood loss and operative time were similar in primary excision and secondary excision (both P>0.05). Histopathologically, both fundus and lateral margins of all the samples were negative. Of the 76 samples, cancer tissue developed outside the mucosal layer in 37 samples, infiltrated into the submucosal layer (pT1 stage) in 33 samples, and infiltrated into the muscular layer (pT2 stage) in 6 samples; 57 samples were classified as grade G1 and 19 samples were classified as grade G2, respectively. The operative complication rate was 6.7%(6/90). The mean postoperative hospital stay was (3.0±1.5) d. No recurrence was noted during the follow-up (median 3.9, 0.4 to 10.0 years).

CONCLUSIONSTEM can be the preferred option for complete removal of middle-upper small (<2 cm) rectal NET(G1-2). For rectal NET with incomplete resection by colonoscopic polypectomy, the secondary TEM can still obtain ideal efficacy even though operative difficulty increases.

Objective To investigate the independent association of the body mass index(BMI) with serum uric acid among shorter children and adolescent. Methods A cross-sectional study was conducted to collect general clinical data of 922 subjects with shorter statures. In each case, their information about height, weight, insulin-like growth factor-Ⅰ(IGF-Ⅰ), growth hormone peak, uric acid, and blood lipids were collected, and their BMI and BMI standard deviations score( SDS) were calculated. Smooth curve fitting and multiple piecewise linear regression were used to analyze the relationship between BMI SDS and uric acid. Results Univariate analysis found that serum uric acid was positively correlated with age, height standard deviation score, body weight standard deviation score, BMI SDS, IGF-Ⅰ standard deviation score, growth hormone peak, triglyceride, and creatinine ( P＜0. 05 ), while negatively correlated with high-density lipoprotein-cholesterol( P＜0.01). Female had significantly lower serum uric acid levels than male(P=0.003). The serum uric acid level was significantly increased after puberty(P＜0.01). After adjusting for potential confounders, the smooth curve fitting revealed a U-shaped relationship between BMI SDS and uric acid, with a BMI SDS inflection point of-0.7. In the population of BMI SDS＜-0.7, for every unit increase of BMI SDS, serum uric acid decreased by 11. 78 μmol/L( P=0. 030). In the population of BMI SDS≥-0. 7, uric acid increased by 11.79 μmol/L(P＜0.01) for every unit increase of BMI SDS. Conclusion Serum uric acid levels seem to be affected by BMI, and increased in population with thin or obesity. We should pay attention to the measurement of serum uric acid in both thin and obese children.

For colorectal surgeons, how to reduce anastomotic leakage after laparoscopic rectal cancer surgery remains to be challenging. We provide a brief discussion regarding the surgical skills required to prevent anastomotic leakage after rectal cancer surgery, such as the following: 1) Low ligation of inferior mesenteric vessel during laparoscopic total mesorectal excision can improve anastomotic tension and blood supply, thus reducing the risk of anastomotic leakage.While high ligation of inferior mesenteric artery resultsin poor blood supply and high tension in atastomotic site, thus increasing the risk of anastomotic leakage. 2) Protective enterostomy is recommended for patients with high risk of developing anastomotic leakage. 3) Use of abdominal/pelvic drains after colorectal anastomosis is recommended to decrease the incidence of anastomotic leakage, early detect anastomotic leakage, and conservativdy manage anastomotic leackage through drainage of pelvic effusion. 4) Laparoscopic reinforcing sutures should be used if anastomotic tension and blood supply are unsatisfactory, including continuous suture with 3-0 or 4-0 absorbable suture and 2-needle interrupted suture in the weak anastomosis. However, these sutures should be performed by experienced surgeons. For male patients with narrow pelvis and those with low rectal cancer, laparoscopic reinforcing sutures should be performed carefully due to the limited operative space. 5) Intraoperative air leak test is recommended to identify the anastomotic integrity for those with suspicious mechanically insufficient rectal anastomosis. 6) Experienced surgeon can reduce the incidence of anastomotic leakage after rectal cancer operation.

Objective To analyze the expressions of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) in the peripheral blood of patients with active tuberculosis (ATB ) or latent tuberculosis infection (LTBI) ,and to evaluate its diagnostic value in differentiation of ATB and LTBI .Methods Forty-eight patients including 18 ATB cases and 30 LTBI cases were continuously enrolled from Wuxi No . 5 People′s Hospital and Huashan Hospital affiliated to Fudan University from January 2011 to March 2013 .Flow cytometry was applied to detect the CTLA-4 expression in CD4+CD25+ FoxP3+ T cells in the peripheral blood of the 48 subjects .CTLA-4 levels were compared using non-parametric Mann-Whitney U test .Results The median percentage of CTLA-4+ Treg in CD4+ CD25+ Foxp3+ Treg cells of ATB patients was 18 .95％ (quantile range :13 .86％ ,27 .73％ ) ,and that in LTBI patients was 6 .67％(quantile range :5 .74％ ,9 .59％ ) ,which was statistically significant (U=18 .0 , P< 0 .01) .Receiver operating curve (ROC) based on the CTLA-4 expression indicated that the area under the curve was 0 .96 , with the optimum cut-off value of 13 .25％ .Thus ,the sensitivity and specificity for the diagnosis of ATB were 86 .7％ and 94 .4％ ,respectively .Conclusion CTLA-4 has highly sensitivity and specificity for the differential diagnosis of ATB and LTBI whose interferon-gamma releasing assays are all positive ,which may also provide meaningful clue for the study of pathogenesis of ATB .