Objective To study the effects of neferine on the expression of glutathione S-transferase-pi in vitro and to explore the multi-drug resistance reversing mechanism of neferine.Methods 50% inhibition concentration(IC_(50)) of ADM on K562/A02 was determined by MTT method.The transcription of GST-? gene was detected by semi-quantitative RT-PCR and the expression level of GST-? was determined by Western blot after neferine treatment.Results Neferine remarkably enhanced chemosensitivity to ADM of K562/A02 cells.After neferine treatment in day 1,day 3 and day 5,the relative efficiency of K562/A02 to ADM was 9.6%,41.4% and 10.7%,respectively.Semi-quantitative RT-PCR showed that mRNA transcription of GST-? gene was significantly reduced(P

Object To investigate the effect of Angelica polysaccharide (APS) on the induction of chronic myelocytic leukemia cells into chronic myelocytic leukaemia dendritic cells (CML-DCs). Methods Bone marrow monocytes from CML patients were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4 combined with APS in each concentration (50, 100, 200 mg/L), respectively. The morphotype of CML-DCs was identified by optical microscope or electron microscope, CML-DCs viability was calculated by Trypan Blue exclusion. The phenotype of CML-DCs (CD 80, CD 86, and CD 83) was identified by flow cytometry. The capability of stimulating auto-lymphocyte or allo-lymphocyte proliferation was tested with mixed leukocyte reaction (MLR). Results Bone marrow monocytes from CML patients, which were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4/APS showed typical morphotype and expressed the high level phenotype of CML-DCs. The capability of proliferation and the survival rate of CML-DCs were enhanced markedly and the expression of CD 83, CD 80, and CD 86 on CML-DCs were significantly increased when CML-DCs were cultured in GM-CSF/IL-4/APS. The capability of stimulating lymphocyte proliferation was more competent in 100 mg/L APS group. Conclusion The expression of CD 83, CD 80, and CD 86 on CML-DCs cultured in GM-CSF/IL-4/APS is significantly higher than those in GM-CSF/IL-4. The capability of CML-DCs of stimulating lymphocyte proliferation is more potential in GM-CSF/IL-4/APS than in GM-CSF/IL-4. APS can promote the induction and mature of CML-DCs cultured in IL-4 and GM-CSF.

Objective To study long-term outcome and prognostic factors of nasopharyngeal carcinoma treated by intensity modulated radiotherapy.Methods A total of 299 patients with nondisseminated nasopharyngeal carcinoma who received initial radiotherapy were analyzed retrospectively.The primary lesion and the upper neck received 70 Gy (5 fraction per week in all 30 fraction) by intensitymodulated radiotherapy (IMRT).The lower neck and the supraclavicular fossa was given 54 Gy (5 fraction per week in all 30 fraction) by a single anterior tangent field with spinal cord block.A median dose of 9.2 Gy (4-20.Gy) was given to the residual primary lesion by IMRT or X-knife.The Kaplan-Meier method was used for calculating the overall survival (OS),disease progression-free survival (DPFS),distant metastasisfree survival (DMFS),Log-rank test was used for evaluating the differences between groups.Multivariate prognostic factor was analyzcd by Cox method.Results The follow-up rate was 99.7％.119 patients were followed-up more than with 5 years.The 5-year OS for stage Ⅰ + Ⅱ,stage Ⅲ and stage Ⅳ were 97.1％,82.7％ and 52.2％(x2=46.19,P=0.000),the 5 years DPFS were 100％,77.6％ and 57.7％ (x2=23.29,P =0.000),DMFS were 100％,82.3％,63.7％ (x2 =16.57,P =0.000) respectively.The 5 year OS,DPFS and DMFS of male and female were 70.7％ vs 94.1％ (x2=16.82,P=0.000),71.5％ vs 87.3％ (x2 =4.74,P =0.029) and 77.2％ vs 89.7％ (x2 =4.38,P =0.036) respectively.For patients who were younger than 45-years,the male had a significantly unfavorable 5-year OS (66.8％ vs.91.2％,x2=7.07,P=0.008),DPFS (59.9％ vs.91.2％,x2=7.72,P=0.005) and DMFS (66.4％ vs.94.0％,x2 =8.46,P =0.004) ;For patients who were old than 45-years,only OS was significantly different between male and female (72.2％ vs.96.0％,x2 =10.19,P =0.001).Multivariate analysis showed the independent prognosticfactors for OS,DPFS,DMFS,were gender (x2 =14.27,5.72,17.64,P =0.000,0.017,0.000),TNM stage (x2 =5.33,15.70,10.57,P =0.021,0.000,0.001) and lymph nodes capsular invasion (x2 =4.30,11.08,21.24,P =0.038,0.001,0.000).Intracranial invasion and supraclavicular lymph node metastasis were independent prognostic factors for OS (x2 =13.32,5.38,P =O.000,0.020).Conclusions The TNM stage,lymph nodes capsular invasion and gender are independent prognostic factors for nasopharyngeal carcinoma treated by intensity-modulated radiotherapy.The patients of younger than 45years own a worse outcome.

Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia (AA), high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ± 0. 29, 2. 26 ± 0. 37 and 1.58±0. 33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively (P < 0.01). The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0. 11,respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.

OBJECTIVE: To determine the effect of neferine (Nef) combined with mdr-1shRNA on the expression of mdr/P-gp in K562/A02 cell line. METHODS: MTT assay was used to observe the cell proliferation. The expression level of P-gp was determined by Western blot and the transcription of mdr-1 gene was detected by semi-quantitative RT-PCR. RESULTS: After K562/A02 cells were treated by Nef or mdr-1shRNA alone or both for 24 h, the proliferation of K562/A02 cells was significantly higher in the Nef combined with mdr-1shRNA treatment group than that of Nef or mdr-1shRNA alone group (P<0.01).The expression of mdr-1/P-gp in the Nef with mdr-1 shRNA group was significantly lower than that of Nef or mdr-1shRNA alone group. CONCLUSION Nef enhances the inhibition of mdr-1shRNA expression vector on K562/A02 cell proliferation and on P-gp protein to effectively reverse multidrug resistance induced by mdr-1 gene encoding P-gp.
ATP Binding Cassette Transporter, Subfamily B ; ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; metabolism ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; K562 Cells ; RNA, Small Interfering ; genetics ; pharmacology

Objective:To explore the effect of ultrasound-guided femoral nerve block with dexmedetomidine combined with ropivacaine on the pain of patients after knee arthroscopy combined with high tibial osteotomy(HTO).Methods:A total of 97 patients who underwent the treatment of HTO combined with knee arthroscopy in the Ankang central hospital from 2021 to 2022 were selected,and they were divided into the observation group(46 cases)and the control group(51 cases)based on the random number table method.All patients received anesthesiology with ultrasound-guided femoral nerve block.For anesthesia,the observation group used ropivacaine combined with dexmedetomidine,and the control group only used ropivacaine.The visual analogue scale(VAS)scores of them were assessed respectively at rests before surgery,and 3h,6h and 12h after surgery,and the times that patients pressed the button of patient controlled analgesia(PCA)within 4 time intervals included the postoperative 0-6h(T1),6-12h(T2),12-18h(T3)and 18-24 h(T4)were recorded.In addition,the doses of oral opioid drug within postoperative 24 h,48 h and 72 h also were recorded,and the relevant adverse reactions of anesthesiology after surgery were recorded.Results:The pain levels at postoperative 3h,6h and 12h in the observation group were significantly lower than those in the control group,and the differences were statistically significant(t=2.181,3.113,3.073,P<0.05),respectively.The oral dosage of opioids within 24h,48h and 72h after surgery in observation group were significantly less than those in the control group,and the differences were statistically significant(t=6.356,9.778,11.284,P<0.05),respectively.The times of pressing PCA button within T1 and T2 intervals after surgery in observation group were also significantly lower than those in control group,and the differences were statistically significant(t=5.033,2.184,P<0.05),respectively.The incidence of adverse reactions in the observation group was significantly lower than that in the control group,and the difference was statistically significant(x2=4.562,P<0.05).Conclusion:In the surgical anesthesia of knee arthroscopy combined with(HTO),the early analgesic effect of femoral nerve block with dexmedetomidine combined with ropivacaine is significant,and the oral dosage of opioids of patient decreases after surgery,and the risk of occurring adverse reaction that is relative to anesthesia after surgery is less.

Composite lymphoma (CL) involving B-cell lymphoma and T-cell lymphoma is extremely rare. Herein, we report three such cases using immunohistochemistry, flow cytometry, and the next-generation sequencing (NGS) to identify the pathological and molecular characteristics of CL. In the first case, the patient was admitted to hospital for generalized pruritic maculopapular rash over the whole body. An excisional biopsy of the skin lesions showed T-cell lymphoma. At the same time, the staging bone marrow (BM) biopsy revealed a diffuse large B-cell lymphoma (DLBCL). After R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) therapies, the patient produced a good response with substantial dissipation of the rashes and relief of skin. The other two patients were admitted to hospital due to lymphadenopathy and were diagnosed with DLBCL and follicular lymphoma (FL) after core needle biopsy of lymph nodes, BM biopsy, BM aspiration, and flow cytometry. Following R-CHOP and R-COP (rituximab, cyclophosphamide, vincristine, and prednisone) therapies, they achieved complete remission unconfirmed (CRu) and complete remission (CR). However, one or two years later, they suffered a relapse of lymphadenopathy. The shocking fact was that re-biopsy of lymphadenopathy revealed peripheral T-cell lymphoma (PTCL) and angioimmunoblastic T-cell lymphoma (AITL). NGS findings identified DNA methyltransferase 3a (DNMT3a), isocitrate dehydrogenase 2 (IDH2), Ras homolog gene family, member A (RHOA), splicing factor 3B subunit 1 (SF3B1), and tumor protein p53 (TP53) mutations. After immunochemotherapy, these patients achieved CRu and CR again. Nevertheless, they suffered a second relapse of T-cell lymphoma. Finally, they died due to progression of disease. We found that the occurrence of CL is associated with Epstein-Barr virus infection and DNMT3a, IDH2, and TP53 mutations, and the prognosis of the disease is closely related to the T-cell lymphoma components.
Humans ; Rituximab/therapeutic use* ; Vincristine/therapeutic use* ; Prednisone/therapeutic use* ; Epstein-Barr Virus Infections/drug therapy* ; Herpesvirus 4, Human ; Neoplasm Recurrence, Local ; Lymphoma, T-Cell/drug therapy* ; Cyclophosphamide/therapeutic use* ; Lymphoma, Large B-Cell, Diffuse/pathology* ; Doxorubicin/therapeutic use* ; Lymphadenopathy/drug therapy* ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use*