Objective:To investigate the effect of tumor-associated macrophage(TAM) on proliferation of renal carcinoma cells and its related mechanism.Methods:The model of TAM was established by stimulating human monocytic leukemia cell line THP-1 with phorbol myristate acetate (PMA), bacterial endotoxin (LPS) and interferon-γ (IFN- γ). Then the TAM model was co-cultured with carcinoma cell lines ACHN and 786-O in vitro .The cytokines IL-6, TNF-α and IL-1β in TAM supernatant were detected by enzyme-linked immunosorbent assay (ELISA). MTT method was used to detect the proliferation of ACHN and 786-O cells treated with supernatant of TAM or TAM/Tocilizumab. Western blot was used to detect lactate dehydrogenase A (LDHA) expression of both renal cancer cells co-cultured with TAM or TAM/Tocilizumab. The ACHN and 786-O cells with LDHA-overexpression and LDHA-knockdown were cultured in TAM supernatant in vitro. The cell proliferation was detected by MTT and the relative proliferation rate was calculated.Results:THP-1 cells was differentiated into TAM through the treatment of 80 ng/ml PMA combined with 20 ng/ml LPS and 20 ng/ml IFN- γ.The expression rate of CD68, a cell surface marker on TAM, was (36.2 ±4.5)%. When TAM was co-cultured with ACHN cells, the results of ELISA showed that the secretion of IL-6 in the supernatant was significantly elevated compared with that in the supernatant when ACHN cells cultured alone [(138.0 ±12.4) pg/ml and (19.7±4.9) pg/ml], and the secretion of TNF- α [(122.5 ±14.2) pg/ml and (12.6 ±2.3) pg/ml] and IL-1 β [(89.2 ±6.4) pg/ml and (69.2 ±3.5) pg/ml] were also significantly increased. The secretion of IL-6 [(119.2 ±14.8) pg/ml and (17.1 ±3.3) pg/ml], TNF- α [(122.6 ±14.4) pg/ml and (45.7 ±7.2) pg/ml] and IL-1 β [(95.1 ±11.8) pg/ml and (88.2 ±12.7) pg/ml] in the supernatant were also significantly elevated when 786-O cells co-cultured with TAM compared with 786-O cells cultured alone. After treated with the supernatant of TAM for 72 hours, the relative proliferation rates of ACHN and 786-O cells [(128.6 ±21.4)% and (124.2 ±19.7)%] were significantly higher than that of the control group (100.0%). At the same time, the expression of LDHA in ACHN and 786-O cells increased significantly. After 72 hours of treatment with the supernatant of TAM combined with tocilizumab, the relative proliferation rates of ACHN and 786-O cells [(76.5±13.7)% and (74.8±12.5)%] were significantly lower than that of the control group(100.0%), and the expression of LDHA was also significantly decreased at the same time. The relative proliferation rates of ACHN and 786-O cells in LDHA overexpression group [(121.5 ±17.2)% and (122.7±21.6)%]were significantly higher than that in blank-vector-transfection group[(93.3±10.7)% and (89.8±11.2)%], while the relative proliferation rates in LDHA-knockdown group [(61.4±11.2)% and (58.0 ±10.6)% ]were significantly lower than that in blank-vector-transfection group.Conclusions:By secreting IL-6, TAM can up-regulate the expression of LDHA and promote the proliferation of renal cancer cells.

Objective:To investigate the clinical efficacy and related adverse reactions of ixazomib-based chemotherapy regimens in the treatment of relapsed/refractory multiple myeloma (RRMM).Methods:Twenty-one patients with RRMM who received ≥2 courses of ixazomib-based chemotherapy regimens in Heze Municipal Hospital and Zoucheng People's Hospital of Shandong Province from October 2018 to February 2020 were collected. Among them, 15 patients had previously received the bortezomib-based regimens, 10 patients had received the lenalidomide-based regimens, and 6 patients had received the treatment regimens containing the above two drugs. The patients were treated by a two-drug or three-drug regimen: 4 mg ixazomib was taken orally on day 1, 8 and 15 in combination with other drugs (dexamethasone, cyclophosphamide or lenalidomide). The therapeutic efficacy and safety were evaluated after the 2nd and the 4th treatment cycles.Results:The overall response rate (ORR) of 21 patients with RRMM after 2 treatment cycles was 38.09% (8/21), including 6 cases of partial remission (PR) and 2 cases of very good partial remission (VGPR). After 4 cycles, ORR was 57.14% (12/21), including 7 cases of PR, 4 cases of VGPR, and 1 case of complete remission (CR). The incidence of grade 3-4 adverse reactions of the ixazomib-based chemotherapy regimens was 23.81% (5/21). Hematological adverse reactions included neutropenia, thrombocytopenia and anemia, and other common adverse reactions included the digestive tract reactions, fatigue, hypokalemia, etc., and the peripheral nerve adverse reactions were all grade 2 or below grade 2.Conclusion:The ixazomib-based chemotherapy regimens are effective and safe in treating RRMM.

Objective To explore the therapeutic efficacy and adverse effects of decitabine combined with half dose CAG regimen and only CAG regimen for the elderly patients with acute myeloid leukemia (AML).Methods A total of 42 elderly patients with AML aged between 65 and 75 years old (except for acute promyelocytic leukemia) admitted into Heze Municipal Hospital from August 2013 to August 2017 were retrospectively analyzed.They were divided into treatment group and control group according to the different chemotherapy regimens.Twenty patients in the treatment group were treated with decitabine combined with half dose CAG regimen (recombinant granulocyte colony stimulating factor + cytarabine + aclarubicin),and 22 patients in the control group were treated with CAG regimen.Results After the first course of treatment,in treatment group,13 cases achieved complete remission (CR),3 cases achieved partial remission (PR),4 cases achieved non-remission (NR),and the total efficacy (CR+PR) rate was 80.0 ％ (16/20).In control group,8 cases (36 ％) achieved CR,2 cases achieved PR,12 cases achieved NR,and the total efficacy rate was 45.5 ％ (10/22).The difference in total effective rate between the two groups was statistically significant (x2 =3.707,P =0.035).There was no significant difference in the bone marrow recovery time,the infusion of red blood cells and platelets between the two groups (all P ＞ 0.05).Conclusion The therapeutic efficacy of decitabine combined with half dose CAG regimen is better than that of CAG regimen,the adverse effects are all tolerated,and it can be served as the prior therapy for elderly AML patients.

Objective To explore the role of B7-H3 in the Tie2 expressing monocytes (TEMs) mediated angiogenesis of clear cell renal cell carcinoma (ccRCC).Methods Level of B7-H3 expression on TEMs surface was detected by flow cytometry in ccRCC tissues and normal renal tissues,which were obtained from April 2016 to August 2016 from 20 patients.Microvessel density (MVD) labeled by CD34 in high B7-H3 + TEMs group and low B7-H3 + TEMs group was detected by immunohistochemical examination in ccRCC specimens.B7-H3 + TEMs and B7-H3-TEMs were co-cultured with the 786-O cell lines,and B7-H3 + TEMs and B7-H3-TEMs culture supernatants were collected as conditioned medium,then the effect of B7-H3 + TEMs on angiogenesis was tested by tubule formation assay and mouse aortic ring assay.Results Flow cytometry showed that the frequency of B7-H3 expression on TEMs in ccRCC was (45.10 ± 17.78)％,and the frequency of B7-H3 expression in normal kidney tissues was (10.28 ± 4.28) ％.The frequency of B7-H3 expression on TEMs was significantly higher than that in normal renal tissues (P ＜ 0.001).The MVD of high B7-H3 + TEMs group (103.81 ± 29.28) was higher than that of low B7-H3 + TEMs group (76.55 ± 20.80) (P =0.027).The results of tube formation assay showed that the number of tubule formation of HUVEC in B7-H3 +TEMs group(55.25 ± 11.48) was significantly greater than that of B7-H3-TEMs group (31.34 ± 8.45) and blank control group (25.00 ± 6.74) (P ＜ 0.001).The results of mouse aortic ring assay showed that the number of neovascularization in B7-H3 + TEMs group(77.35 ± 18.47) was significantly greater than that of B7-H3-TEMs group (39.42 ± 8.29) and blank control group (28.79 ± 7.63) (P ＜0.001).Conclusions B7-H3 + TEMs can promote angiogenesis in ccRCC,and might act as an effective target in anti-angiogenic therapy for ccRCC.

Objective To evaluate the efficacy and safety of Sun's tip-flexible ureterorenoscopy (tf-URS) combined with holmium laser lithotripsy in treatment of renal and upper ureteral calculi.MethodsClinical data of 48 patients (29 males and 19 females) with renal and upper ureteral calculi treated by tf-URS with holmium laser lithotripsy in our hospital from November 2015 to May 2016 were retrospectively analyzed.Among 48 patients there were 29 males and 19 females with a mean age of (37.8±12.5) years (21-67), including 31 cases of upper ureteral calculi and 17 cases of renal calculi with a mean stone size of (1.4±0.6) cm (0.8-2.1).The tf-URS was advanced through the ureter under visual direction with rigid mode.When the endoscope reached the ueteropelvic junction, the inner flexible part was extended out to explore the pelvicaliceal stones and then the holmium laser lithotripsy was performed.The therapeutic results were evaluated by KUB 2 days after operation and color ultrasonography or CT 4 weeks after operation.Results The success rate of insertion of tf-URS was 94% (45/48).The calculi were detected in 45 cases, and laser lithotripsy succeeded in 43 cases (96%).The rate of total stone clearance was 87% (39/45).The procedure was transformed to laparoscopic or percutaneous nephrolithotomy (PCNL) in 3 patients with ureterostenosis or ureteral distortion.No serious complications occurred.Conclusion The tf-URS with holmium laser in the treatment of renal and upper ureteral calculi is safe,reliable and effective.

OBJECTIVE: To analyze the association between genetic polymorphisms of DNA repair genes of XPD (751 Lys/Gln), XPC (PAT)and susceptibility to laryngeal carcinoma. To explore the effect between DNA repair genes of XPD (751 Lys/Gln), XPC (PAT) and carcinogenesis of LSCC(laryngeal squamous cell carcinoma). METHOD: A case-control study was conducted involving 233 LSCC patients and 102 healthy controls to investigate the association between polymorphisms of XPD(751 Lys/Gln), XPC (PAT) and LSCC. All blood samples of the Han people from the Guang Dong Zone was analysze with methods of PCR, PCR-RFLP, ASA and the technique of checking DNA sequencing with sequenator. We explored the association between polymorphisms and the clinical pathologic characteristic of LSCC. The data was compute with SPSS13.0. Odds Ratios (ORs) with 95% CI for relevancy intensity were calculated using binary logistic regression analysis. REULT: There is no difference of the frequency of XPC-PAT and XPD (751 Lys/Gln) genotype between in LSCC and in healthy contradistinguish (P > 0.05). CONCLUSION There may be no association between the susceptibility to laryngeal carcinoma and the genotype of XPC-PAT and XPD (751 Lys/Gln).
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; DNA Repair ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Genotype ; Humans ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Xeroderma Pigmentosum Group D Protein ; genetics

Objective To evaluate the feasibility of bladder extracellular matrix (BACM) seeding with autologous cultured vaginal smooth muscle cells (VSMC) repaired rabbit vaginal defect.Methods This study included 24 female rabbits.BACM and vaginal acellular matrix (VACM) were obtained from 8 rabbits by decellularization process.The other 16 rabbits were randomly divided into experimental and control groups.Vaginal tissue biopsies ( ～ 1 cm2) were harvested from female New Zealand rabbits.VSMC were cultured and stained with a-smooth muscle actin antibodies. Cultured VSMC cells were seeded on BACM ( experimental group) or VACM ( control group) at a cell density of 1 × 107 cells/cm2.The cell-seeded matrixes were cultured for 5days.In experimental group of 8 rabbits,a 2 cm segment of vagina was resected and replaced with BACM seeding with VSMC.Then the regenerative segment was studied with histological technique by hematoxylin-eosin staining after 3,6 and 12 weeks postoperative aud vaginography was performed at 12 weeks postoperative.The 8 rabbits in control group underwent the exact same procedure as above but the vaginal defect was repaired with VACM seeding with VSMC,Results The prepared BACM and VACM were transparent,HE staining and scanning electron microscopy showed the two acellular matrix was both consisted of abundant network of fibers with the regular arrangement,without cellular debris.Primary culture of VSMC was successfully established and passaged,and was uniformly spindle-shaped in the confluent state and showed a characteristic hill and valley'formation.Immunohistochemical staining showed VSMC in culture stained positively with a-smooth muscle actin antibodies.VSMC began to adhere to the BACM 5 hour after implanting and the number of the adhered cells increased with time.Cells gradually expanded and showed the typical morphology of smooth muscle cells.Three weeks after implantation in vivo,the luminal surface of matrix was completely covered by vaginal epithelial tissue,a layer of smooth muscle cells was formed in the outer surface of the matrix.Multilayered vaginal epithelial and improved development of organized muscle bundles was observed after 6 weeks.The regenerative tissue was equivalent to the normal vaginal tissue at 12 weeks postoperatively.Vaginography demonstrated the maintenance of full patency and a wide vaginal caliber without fibrosis and graft rejection.There was no significant difference in all evaluated items between experimental and control groups.Conclusions BACM has the same regenerative process as VACM in the replacement of vaginal defect.Moreover,BACM has wider source and appears to be an suitable material for vaginal replacement.

OBJECTIVE: To analyze the association between genetic polymorphisms of xenobiotic- metabolizing enzymes GSTM1, GSTT1, GSTP1 and susceptibility to laryngeal carcinoma from the Han people in Guangdong zone. METHOD: A case-control study was conducted involving 233 LSCC (laryngeal squamous cell carcinoma) patients and 102 healthy controls to investigate the association between polymorphisms of GSTM1, GSTT1, GSTP1 (Ile/Val) and LSCC from the Han people in Guangdong zone. All blood samples of the Han people from the Guangdong zone was analyzed with methods of PCR, ASA and the DNA sequencing technique with sequenator. We explored the association between polymorphisms and the clinical pathologic characteristics of LSCC. The data was processed with SPSS13.0. Odds Ratios (ORs) with 95% CI for relevancy intensity were calculated using binary logistic regression analysis. RESULT: The frequency of GSTM1(-) and GSTT1(-) genotype was higher in LSCC than that in healthy controls (OR = 2.61, 3.05, P < 0.01). There was synergic effect between GSTT1 (-) genotype and heavily smoking during carcinogenesis of LSCC (OR = 3.51, 95% CI 2.05-5.01; OR = 2.99, 95% CI 2.00-4.49). The frequency of GSTM1(-) and GSTT1(-) genotype was higher in LSCC whose family had carcinoma history. The frequency of advanced LSCC was higher in patients who were with GSTM1(-) and GSTT1 (-) genotype (P < 0.05). There was no difference of the frequency of GSTP1(I le/Val) genotype between and in healthy controls (P > 0.05). CONCLUSION There may be an association between the susceptibility to carcinoma and GSTT1(-), GSTM1(-) genotype. The GSTT1(-) polymorphism c gene cooperating with heavily smoking boost up the susceptibility of individual to laryngeal carcinoma. The GSTM1(-) polymorphism c may not cooperating with smoking during carcinogenesis of LSCC in the Han people in Guangdong zone. The morphisms of GSTT1 and GSTM1 gene may affect the carcino-genesis of LSCC in the Han people in Guangdong zone. There may be no association between the susceptibility to laryngeal carcinoma and the GSTP1(Ile/Val) type.
Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Laryngeal Neoplasms ; epidemiology ; ethnology ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic

Objective To evaluate the clinical characters, management and prognostic factors of patients with differentiated invasive thyroid carcinoma (DITC). Methods The data were analyzed retrospectively for 114 DITC patients treated at Department of Head and Neck Surgery of Sun Yat-sen University Cancer Center. Survival analysis was performed by Kaplan-Meier method, comparison among/between groups was performed using log-rank test, and multivariate analysis was carried out using Cox proportional hazard model. Results After surgery, 68 patients were with tumor residue. The 5-year and 10-year overall survival rate were 91.9% and 80.1% respectively in all patients, while the 10-year overall survival rate were 88.5% 、78.5% and 53.1% in no tumor residue group, micro-residue group and grossresidue group respectively. This study failed to prove that radiotherapy might improve the survival rate in patients with postoperative tumor residue. Multivariate analysis indicated that age, invasion to esophagus and recurrence predict the prognosis. Conclusion DITC may be treated mainly by surgical operation. Radical resection is the key factor in the treatment of DITC. Patients with DITC have a relatively poor prognosis.Age, esophagus invasion and status of tumor residue are the most important factors affecting the prognosis.

Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.