Objective:To discuss the effect of urolithin C(UC)on the proliferation,apoptosis,and autophagy of the acute myeloid leukemia(AML)HL-60 cells,and to clarify its mechanism.Methods:The HL-60 cells were divided into different concentrations(20,40,60,80,and 100 μmol·L-1)of urolithin A(UA)groups,urolithin B(UB)groups,and UC groups.CCK-8 assay was used to detect the proliferation activity of the cells in various groups;the morphology of the cells in different concentrations of UC groups was observed under optical microscope.The HL-60 cells were divided into different concentrations(0,20,40,and 80 μmol·L-1)of UC groups and 3-methyladenine(3-MA)combined with different concentrations(0,20,40,and 80 μmol·L-1)of UC groups.CCK-8 assay was used to detect the proliferation activities of the cells in various groups.The HL-60 cells were divided into control group(0 μmol·L-1)and different concentrations(20,40,and 80 μmol·L-1)of UC groups.The live/dead cell staining method was used to detect the dead rates of the cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups;the autophagy of the cells was detected by autophagy staining kit(monodansylcadaverine,MDC)method;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Beclin 1,autophagy related gene 9(ATG9),and autophagy related gene 7(ATG7)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of cysteinyl aspartate specific proteinase-3(Caspase-3),cleaved cysteinyl aspartate specific proteinase-3(Cleaved Caspase-3),microtubule-associated protein 1 light 3(LC-3),extracellular regulated protein kinases(ERK),phosphorylated ERK(p-ERK),AMP-activated protein kinase(AMPK),and phosphorylated AMPK(p-AMPK)in the cells in various groups.Results:The CCK-8 assay results showed that after cultured for 24,48,and 72 h,compared with 0 μmol·L-1 UA,UB,and UC groups,the proliferation activities of the cells in different concentrations of UA,UB,and UC groups were decreased(P＜0.01)with a concentration-and time-dependent manner;at 48 h,compared with UA and UB,the half-maximal inhibitory concentration(IC50)of UC was the lowest.The cell morphology observation results showed that compared with control group,the intercellular connection and the number of the cells were decreased with the increasing of UC concentration,and the cell fragment was increased.The CCK-8 assay results showed that compared with 40 and 80 μmol·L-1 UC groups,the proliferation activities of the cells in 3-MA combined with 40 and 80 μmol·L-1 UC groups were increased(P＜0.05 or P＜0.01).The live/dead cell staining results showed that compared with control group,the dead rates of the cells in 40 and 80 μmol·L-1 UC groups were increased(P＜0.01).The flow cytometry results showed that compared with control group,the apoptotic rate of the cells in 80 μmol·L-1 UC group was increased(P＜0.01).The MDC method results showed that with the increasing of UC concentration,the green fluorescence in the cells in different concentrations of UC groups was gradually intensified.The RT-qPCR results showed that compared with control group,the expression levels of Beclin 1,ATG9,and ATG7 mRNA in the cells in 80 μmol·L-1 UC group were increased(P＜0.01).The Western blotting results showed that compared with control group,the expression levels of Cleaved Caspase-3 protein in the cells in 20,40,and 80 μmol·L-1 UC groups were increased(P＜0.01),the ratio of membrane LC3/cytoplasmic LC3(LC3-Ⅱ/LC3-Ⅰ)in the cells in 80 μmol·L-1 UC group was increased(P＜0.05),and the ratios of p-AMPK/AMPK and p-ERK/ERK in the cells in 40 and 80 μmol·L-1 UC groups were increased(P＜0.01).Conclusion:UC can inhibit the proliferation of the AML HL-60 cells,induce the apoptosis and autophagy,and increase the phosphorylation levels of ERK and AMPK proteins in the cells.

Objective To investigate the risk factors of complications after CT-guided percutaneous transthoracic needle biopsy(PTNB)in the plateau environment.Methods A total of 858 patients who underwent CT-guided PTNB were selected,and the clini-cal data of patients,imaging features of lesions,information related to puncture operation,complications,and pathological results were analyzed retrospectively,then the independent risk factors of postoperative pneumothorax and pulmonary hemorrhage were summarized.Results Among 858 patients with lung biopsy,816 cases(95.1％)were successfully sampled,including 203 cases(23.7％)in the pneumothorax and 140 cases(16.3％)in the pulmonary hemorrhage.The statistical analysis results of the pneumo-thorax revealed significant differences in lesion location and lesion size(P＜0.05).The statistical analysis results of the pulmonary hemorrhage showed significant differences in lesion location,lesion size,puncture angle and puncture depth(P＜0.05).Independent risk factors affecting pneumothorax and pulmonary hemorrhage were illustrated in a forest plot.Conclusion Because the oxygen partial pressure and climate temperature are relatively low in the plateau environment,the cardiopulmonary function of patients will be affected by the living environment.Therefore,on the premise of ensuring the success rate of CT-guided PTNB,the optimal path and timing should be selected to reduce the risk of complications.

Objective To investigate the efficacy of CT-guided percutaneous microwave ablation for the treatment of primary hepatocellular carcinoma.Methods A total of 132 patients with primary hepatocellular carcinoma were divided into control group and study group(66 cases in each group)according to different treatment plans.The control group received transcatheter arterial chemoembolization(TACE)treatment,while the study group received TACE combined with percutaneous microwave ablation under CT guidance.The changes in serum tumor markers and liver function indicators were observed before and after treatment in the two groups,and the efficacy(short-and long-term)and safety of the two groups were compared.Results The levels of serum carcinoembryonic antigen(CEA),alpha-fetoprotein(AFP),carbohydrate antigen 125(CA125),and carbohydrate antigen 19-9(CA19-9)in both groups decreased significantly after treatment compared to those before treatment,and intergroup comparison showed that the levels of CEA,AFP,CA125,and CA1 9-9 in the study group were significantly lower those after treatment(P＜0.05).Compared with those before treatment,the levels of alanine transaminase(ALT)and aspartate transaminase(AST)were decreased,and the level of albumin(ALB)was increased of both groups after treatment.The intergroup comparison showed that the study groups ALT and AST levels were lower and ALB level was higher(P＜0.05).The total effective rate of the study group was clearly higher than that of the control group(75.76％vs 46.97％,P＜0.05).The 1-year survival rates of the two groups were similar(90.91％vs 81.82％,P＞0.05),however,the 2-year survival rate of the study group was clearly higher than that of the control group(84.85％vs 63.64％,P＜0.05).Conclusion CT-guided percutaneous microwave ablation for the adjuvant TACE treatment of primary hepatocellular carcinoma can effectively reduce tumor burden and lower tumor marker levels,its short-and long-term efficacy is significant,with a low incidence of adverse reactions and good safety.

Objective:To establish a sensitive and rapid nucleic acid detection method for 2019-nCoV that can be promoted in primary laboratories.Methods:The detection method for 2019-nCoV nucleic acid was established based on the Bubble Mediated Nucleic Acid Denaturation (BMD)-PCR technology with 2019-nCoV specific primers and probes. To validate this method, the laboratory-confirmed clinical samples (including 60 positives and 68 negatives of 2019-nCoV) were used and meanwhile the result were compared with the commercialized 2019-nCoV nucleic acid detection kits. The specificity of this method was validated by using 11 respiratory viruses and enteroviruses which cover common genotypes.Results:The BMD-PCR method showed good specificity in the detection of 2019-nCoV nucleic acid. It had no cross-reactivity with 11 common respiratory viruses and enteroviruses, and the limit of detection was less than 600 copies/ml. In addition, the qualitative result of 128 clinical samples were 100% consistent with the commercialized detection kits. Furthermore, the detection time can be shortened to 28 minutes.Conclusions:BMD-PCR for 2019-nCoV nucleic acid rapid detection could quickly diagnose and identify clinical samples, and could meet the current urgent needs of large-scale population and high-throughput screening which has important public health significance.

Objective To analyze the clinical and biological characteristics of adult acute T-lymphocytic leukemia (T-ALL) patients carrying SET-NUP214 fusion gene,and the prognostic value of SET-NUP214 molecular marker monitoring.Methods The clinical and laboratory data of 4 adult T-ALL patients with SET-NUP214 fusion gene in the Third People's Hospital of Chengdu from January 2009 to December 2014 were analyzed retrospectively,and T cell receptor (TCR) gene rearrangement was detected to judge the differentiation and developmental stages of tumor cells in these patients.Minimal residual disease (MRD) was detected in 2 patients with follow-up specimens through detection of SET-NUP214 gene by using polymerase chain reaction.Results Four patients expressed T cell immune markers CD5,CD7,and cytoplasmic CD3 (cyCD3),and also expressed some myeloid-specific antigens.All 4 patients had the same SET-NUP214 fusion site.In the tumor cells of 4 patients,5 TCRB gene rearrangements were detected,all of which were incomplete rearrangement of DB-JB;4 patients were detected with TCRG and TCRD gene rearrangements,and all were completely rearranged.The result of MRD monitoring through SET-NUP214 fusion gene was consistent with clinical treatment outcome.Conclusions The T-ALL patients with SET-NUP214 fusion gene have some unique cell biological characteristics.SET-NUP214 fusion gene could be used as a molecular marker for MRD monitoring,and which can be used for the follow-up in the course of treatment.

OBJECTIVE： To prepare Zingiber officinale oil microcapsules and to evaluate its quality. METHODS： Z. officinale oil microcapsules were prepared by spray drying method with sodium starch octenyl succinate as capsule material. The preparation technology was optimized by orthogonal test with mixing temperature of capsule material and capsule core， mass ratio of capsule material and capsule core， stirring speed as factors， using encapsulation efficiency as index. The drug loading， encapsulation efficiency， appearance， particle size distribution and stability of light， heat and humidity （using iodine value and peroxide value as indexes） were evaluated. RESULTS： The optimal preparation technology of Z. officinale oil microcapsules was that the mixing temperature of capsule material and core was 60 ℃； mass ratio of capsule material and capsule core was 10 ∶ 1； stirring speed was 12 000 r/min. Average drug-loading amount and encapsulation efficiency of Z. officinale oil microcapsules prepared by optimal technology were 17.97％ and 73.57％ （n＝3）. The morphology of Z. officinale oil microcapsules was round， smooth， non-sticky and uniform in size distribution. The average diameter of microcapsules was （6.30±0.27） μm. Under light， heat and humidity conditions， the iodine value and peroxide value of Z. officinale oil microcapsules changed slightly. CONCLUSIONS： The optimal preparation technology of Z. officinale oil microcapsules is simple and reproducible. The prepared microcapsules have good encapsulation efficiency， high drug loading amount and good stability.

Objective To evaluate the diagnostic value of breast mass-like lesions by digital breast tomosynthesis technique (DBT) versus full field digital mammography (FFDM).Methods 182 breast mass cases undergoing respectively DBT and FFDM diagnosis were reviewed to evaluate the the sensitivity,specificity and accuracy,breast BI-RADS classification and differences of edge character.Results Of the 182 cases,101 cases were malignant,81 cases were benign.DBT and FFDM in malignant mass detection rate were 95.0％ and 95.0％ respectively,the benign tumor detection rate were 80.2％ and 80.2％ respectively,the difference was statistically significant (P ＜ 0.05).The diagnostic sensitivity was 93.1％ (94/101) and 82.2％ (83/101),specificity was 66.7％ (54/81) and 53.1％ (43/81),accuracy was 81.3％ (148/182) and 69.2％ (126/182),all the differences were statistically significant (P ＜ 0.05).The BI-RADS classification difference of the malignant mass was statistically significant (x2 =12.912,P =0.044 5),and the benign mass was also statistically significant (x2 =12.739,P =0.026 0).The clear edge benign tumors detected by DBT and FFDM respectively were 65 and 45 cases (x2 =10.224,P =0.001 4).The spicule sign detected by DBT and FFDM respectively in malignant tumors were 71 and 50 cases (x2 =8.244,P =0.004 1).Conclusion DBT compared to traditional FFDM photography improves the lesion visibility,increases the diagnostic sensitivity and specificity,conducing to the identification of benign and malignant lesions.

Objective To investigate the potential therapeutic efficacy of lentivirus-mediated artemin (ARTN) gene modified bone marrow mesenchymal stem cells (MSCs) transplantation on the rat model of Parkinson' s disease (PD) and the effects on expression of brain-related proteins.Methods MSCs were isolated and cultured in vitro,transfected by recombinant lentiviral vectors carrying ARTN gene.The PD rat model established by 6-hydroxydopamine (6-OHDA) was randomly divided into 5 groups:Sham group,PD group,MSCs group,MSCs transfected with empty lentiviral vectors transplanted (LV-MSCs)group and MSCs transfected with recombinant lentiviral vectors carrying ARTN gene transplanted (LVARTN-MSCs) group.The MSCs,LV-MSCs and LV-ARTN-MSCs groups were transplanted into the left striatum of each rat model of PD and ethology tests in every group were made with intraperitoneal injection of apomorphine (APO) 2,4,6,8 weeks after transplantation.The expression of tyrosine hydroxylase (TH) protein in substantia nigra (SN) was measured by Western blotting and immunohistochemistry,and immunofluorescence showed ARTN gene modified MSCs expression in rat brain tissue.The levels of dopamine (DA),dihydroxy-phenylacetic acid and homovanillic acid in striatum of each group were detected by high performance liquid chromatography.Results After injection of APO,rotation frequency decreased in LV-ARTN-MSCs group,i.e.(179.33 ± 10.74) circles/30 min vs (235.83 ± 18.95),(203.67 ±11.50) and (206.33 ± 11.86) circles/30 min in PD,MSCs and LV-MSCs groups (q =8.828,P ＜ 0.01;q =3.802,P ＜ 0.05;q =4.219,P ＜ 0.05).The percentage of TH-positive cells in SN after cell transplantation was increased significantly in LV-ARTN-MSCs group (64.05％ ± 5.49％) when compared with PD group (34.18％ ±3.35％),MSCs group (52.59％ ±4.48％) and LV-MSCs group (50.57％ ± 4.41％),respectively (q =13.280,5.135,6.028,all P ＜0.01).At the same time,TH protein in SN after cell transplantation was also increased obviously in LV-ARTN-MSCs group.ARTN gene modified MSCs can survive for at least 6 weeks in the rat brain of PD,mainly concentrated in the transplantation side of striatum.Eight weeks later,the levels of DA in striatum after cell transplantation were elevated significantly in MSCs group (2.34 ± 0.54),LV-MSCs group (2.28 ± 0.45) and LV-ARTN-MSCs group (2.28 ± 0.45)when compared with PD group (0.87 ± 0.07) (q =5.233,P ＜ 0.05;q =5.020,P ＜ 0.01;q =20.190,P ＜ 0.01),and LV-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN gene modified bone marrow MSCs transplanted into the striatum of brain may have therapeutic effects on rat models of PD.

Objective To discuss the protective effect of Artemin (ARTN) on 6-hydroxydopamine (6-OHDA)-mediated neurotoxic injury,and the alternations of related proteins in Parkinson's disease (PD) model rats.Methods Bone marrow mesenchymal stem cells (MSCs) were isolated and cultured in vitro.After being transfected with recombinant lentiviral vectors carrying A RTN gene,MSCs stably expressed ARTN were chosen (Lv-ARTN-MSCs cells).The SH-SY5Y cells were treated with supematant of Lv-ARTN-MSCs prior to 6-OHDA treatment,and then,cell survival rate was measured by MTT assay and morphologic changes in cultured SH-SY5Y cells were observed by fluorescence microscope (Hochest33258 staining).PD rat models were established and randomly divided into four groups (n=6):PD group,MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group; and sham-operated group (n=6) was also chosen.The PD,MSCs,Lv-MSCs and Lv-ARTN-MSCs groups were transplanted with 5 μL of saline,MSCs (1.0×l05 cell/5 μL),empty virus modified MSCs (1.0×105 cell/5 μL) and ARTN gene modified MSCs (1.0×105 cell/5 μL),respectively,into the left striatum; rats in the sham-operated group were injected with saline when rats in the other groups were received 6-OHDA injection (the same surgical procedures and coordinates).The expressions of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the striatum were measured by Western blotting.Results Western blotting indicated obvious ARTN protein expressions in the Lv-ARTN-MSCs groups.The supematant of Lv-ARTN-MSCs could effectively reduce the apoptosis rate induced by 6-OHDA; as compared with that of SH-SY5Y cells in the 6-OHDA group,the cell survival rate in the Lv-A R TN-MSCs group increased by 13.67％,with significant difference (P＜0.05).Eight weeks after transplantation,the levels of TH and DAT protein in the striatum were elevated significantly in MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group as compared with those in the PD group (P＜0.05),and the Lv-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN which is a secreted protein can protect dopaminergic neuron against 6-OHDA-induced toxicities in Parkinson's disease,and the mechanism might be related to the increased expressions of TH and DAT.

OBJECTIVETo investigate the predictive factors affecting sentinel lymph node status in early breast cancer patients.

METHODSClinicopathological data of 1 038 patients with early breast cancer, who underwent sentinel lymph node biopsy in Henan Tumor Hospital between July 2010 and August 2013, were reviewed. Logistic regression analysis was performed to identify the relevance of clinicopathological features with sentinel lymph node metastases.

RESULTSThis group was consisted of 1 038 female patients with an average of 48.6 years. Positive sentinel lymph nodes were found in 22.9% (238/1 038) of the patients. The average number of sentinel lymph nodes removed by surgery was 3.8. Tumor size, tumor location, histopathology, ER/PR status and Ki-67 level were significantly correlated with SLN metastasis(P < 0.05 for all). All the above factors but Ki-67 level were significant independent predictors for SLN metastasis(P < 0.01 for all).

CONCLUSIONNegative hormone receptor status, invasive cancer of non-specific histopathological type, tumor size >2 cm, and tumor location in the outer upper quadrat are independent predictive factors of sentinel lymph node metastasis in patients with early breast cancer.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; surgery ; Female ; Humans ; Ki-67 Antigen ; metabolism ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies ; Sentinel Lymph Node Biopsy ; Tumor Burden ; Young Adult