Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

Objective To evaluate the efficacy and safety of 131I-metaiodobenzylguanidine (131 I-MIBG) in treatment of malignant pheochromocytoma/malignant paraganglioma (MPHEO/MPGL).Methods The clinical data of 96 cases of MPHEO/MPGL (60/36) treated with 131I-MIBG between December 1998 and April 2014 were retrospectively reviewed.Among them,the malignant pheochromocytoma was found in 60 cases and malignant paraganglioma was found in 36 cases.Seventy-eight patients (81.2％)presented initially with hypertension,whereas 18 patients (18.8％) presented adrenal incidentaloma.Before 131I-MIBG treatment,24 h urinary norepinephrine was (409.5± 127.2) nmol,24 h urinary dopamine was (99.3±41.1) nmol,24 h urine adrenaline was (1 409.9±336.0) nmol.Before treatment,the compound iodine solution was given to each one.Then,all patients were given an initial course of 131I-MIBG therapy (5.55,7.40 GBq).Subsequent 131I-MIBG treatment (5.55,7.40 GBq) was undertaken every three to six months.The patients got symptomatic,hormonal or radiological response underwent sbsequent 131I-MIBG therapy (3.70,5.55 GBq) every year.All patients underwent clinical symptoms (headache,palpitate,sweating,hypertension),biochemical (24 h urine catecholamin) and radiological evaluation (CT/MRI) within 6 months to evaluate the efficacy and safety of 131I-MIBG treatment.Results After one to eleven sessions of 131 I-MIBG treatment,in total,266 doses of 131 I-MIBG were administered,average dose was 6.49 GBq.22.9％ of patients demonstrated radiological partial response (≥ 50％ reduction in tumor size) after first or repeated 131 I-MIBG treatment.Eleven cases (11.5％) achieved clinical complete response,41 cases (42.7％) achieved clinical partial response and 23 cases (24.0％) maintained the stable clinic symptoms.After treatment,24 h urinary norepinephrine (164.3±71.6) nmol and dopamine (49.7±24.7) nmol showed significantly decline,compared with those before treatment (P＜ 0.05).While,24 h urinary epinephrine (1 354.7±433.4) nmol had no obvious change (P＞0.05).No life-threatening adverse events were reported,but 2 MPGL patients developed transient leucopenia or thrombocytopenia after four and five times 131 I-MIBG treatment,respectively.Conclusions Treatment with repeated low dose 131I-MIBG is well tolerated and effective in controlling the progression and alleviating the clinical symptoms.The 131I-MIBG therapy is an effective and safe treatment modality for MPHEO/MPGL.

Purpose To detect the expression of autophagic genes Beclin 1 and MAP1LC3 in cutaneous malignant melanoma and to ex-plore the relationship between autophagia and malignant melanoma. Methods 85 cases of speicmens including normal skin tissue, in-tradermal nevi, radial growth phase melanomas, vertical growth phase melanomas, and metastatic melanoma were collected, and the protein expression of Beclin 1 and MAP1LC3 were evaluated by immunohistochemistry of SP methods. Results The Beclin 1 and MAP1LC3 expression were pretended to be 100% in normal skin tissue, and they were declined to 85% and 95% in intradermal nevi, 58% and 50% in radial growth phase melanomas, 49. 5% and 44. 4% in vertical growth phase melanomas, both of 17% in melanoma metastases (P<0. 05). Conclusion Beclin 1 and MAP1LC3 autophagic gene expression were significantly decreased with tumor pro-gression, as well as was correlated with conventional histopathologic prognostic factors.

Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24expressing human melanoma differentiation associated gene-7 (mda-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potential of HepG2, SMMC7721, MHCC97L and normal liver cell line LO2. Methods The oncolytic adenovirus SG600-IL24 which carrying mda-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line. The mRNA and protein expression of mda7/IL-24 in HepG2, SMMC7721, MHCC97L and LO2 cell lines was confirmed by RT-PCR,ELISA assay and Western blot respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst33258 and flow cytometry were studied to indicate the apoptosis effects. Results It was confirmed by RT-PCR, ELISA assay and Western-blot that the exogenous mda-7/IL-24 gene was highly expressed in HepG2, SMMC7721, MHCC97L and LO2 cell lines. MTT and apoptosis detection indicated that MDA-7/IL-24 can induce the growth suppression (the inhibition rate was 75% ±2. 5% ,86% ±3. 5% ,and promotes apoptosis ( the apoptosis rate was 56. 5% ± 4. 0% , 34. 4% ± 2. 0% , 43. 3% ± 2. 5%cell lines at G2/M phase ( the blocking rate was 35. 4% ± 4. 2% , 40. 5% ± 5. 0% , 42. 0% ± 5. 0%metastatic potential hepatocellular carcinoma cell lines but not in normal liver cell line.Conclusions Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression, promote apoptosis in hepatocellular carcinoma lines in vitro but not in normal liver cell LO2.

Objective To evaluate the curative effectiveness and safety of human chorionic gonadotropin(HCG) treatment on children with microphallic hypospadias.Methods A total of 48 consecutive children with microphalic hypospadias were enrolled in the study,and the children were randomized into the experiment group(HCG treatment) and the control group with the research randomizer.The patients in experiment group were treated with HCG prior to surgery,and the control group did not received any hormone therapy preoperatively.All children in the experiment group and the control group underwent hypospadias repair by using transverse preputial island flap (Duckett technique) urethroplasty.Penile length,diameter of glans penis,bone age,serum testosterone level,and secondary effects were recorded before and after therapy in the experiment group.Postoperative complications were assessed with respect to fistulas,urethral strictures,diverticula,meatal stenosis,and glanular dehiscence in both groups.Results (1)Mean penile length and diameter of the experiment group increased significantly by (1.08±0.47) cm (t=-5.196,P<0.05) and (0.31±0.06) cm and there was a significant difference between before and after treatment (t=-5.080,P<0.05).(2)Urethrocutaneous fistulas were observed in 8 patients in the control group compared to 2 patients in the experiment group with a statistically significant difference (χ2=4.547,P<0.05).There was a significant difference between the overall reoperation rates of control group (9 patients) and the experiment group (3 patients,χ2=4.000,P<0.05).The penile tissue of the patients in the experiment group was soft and able to be easily separated and released during the operation and the flap had more blood supply.Conclusions Pretreatment with HCG therapy prior to hypospadias repair is beneficial to children with microphallic hypospadias.Significant penile growth was seen in the children treated with HCG and there was no obvious side effect.Moreover,pretreatment with HCG is beneficial to decrease the complications and reoperation rates of hypospadias repair which proves to be effective and safe.

OBJECTIVE:To explore the role of clinical pharmacists on pharmaceutical care for allergic reaction caused by ox-aliplatin. METHODS:Clinical pharmacists conducted pharmaceutical care for a patient with oxaliplatin-induced allergic reaction, and suggested stopping taking oxaliplatin,giving Dexamethasone injection 5 mg and then slowing down injection speed. RE-SULTS:Physicians adopted the suggestions of clinical pharmacists. Allergic reaction relieved 5 min after giving Dexamethasone in-jection. The patient didn't suffered from this allergic reaction again under tight supervision. CONCLUSIONS:Oxaliplatin is often used for tumor therapy. Medical staff should be familiar with the prevention,diagnosis and treatment of ADR,evaluate oxaliplatin chemotherapy plan in advance and screen high risk allergy factor. The participation of clinical pharmacists in pharmaceutical care contribute to ADR monitoring and promote safe and rational drug use in the clinic.

BACKGROUND:Umbilical cord mesenchymal stem cells are plentifully and conveniently obtained with a high proliferative ability, and have opened up a new way to treat patients with liver failure as they can differentiate into hepatocyte-like cells.OBJECTIVE:To observe the safety and efficacy in the treatment of chronic liver failure by transplanting umbilical cord mesenchymal stem cells.METHODS:Using parallel contrast method, 50 patients with chronic liver failure were divided into two groups, namely a stem cell group and a control group, containing 25 patients in each group. For the first group, transplantation of umbilical cord mesenchymal stem cells, (1.4-2.3)×106/kg, 100 mL, was given on the basis of medical comprehensive treatment,while for the second group only simple medical comprehensive treatment was given. The injection was done every 15 days, totally three times. Liver functions, prothrombin activity, Model for End-Stage Liver Disease (MELD) score, clinical symptoms, survival and side effects of the patients were observed before and 2, 4, 12 and 24 weeks after the treatment.RESULTS AND CONCLUSION:Compared with the control group, the albumin level and prothrombin activity were significantly increased in the stem cell group 12 and 24 weeks after treatment (P < 0.05), while the MELD score was significantly decreased in the stem cell group at 4, 12 and 24 weeks after treatment (P < 0.05). There was no significant difference in alanine aminotransferase and total bilirubin levels between the two groups (P > 0.05). Four weeks after treatment, clinical symptoms of the stem cell group improved significantly in comparison with the control group (P < 0.05).During the 24-week follow-up, the survival rate in the stem cell group was significantly higher than that in the control group (P < 0.05). Additionally, there were no adverse reactions and liver cancer associated with the stem cell therapy.Results show that it is safe and effective to treat patients with chronic liver failure through the transplantation of umbilical cord mesenchymal stem cells, and the cell transplantation can significantly improve patients' survival rate.

Objective To investigate the expression and significance of MRP3 in pancreatic cancer and different tumor cells. Methods MRP3 expression was detected in 30 cases of pancreatic cancer tissues and the adjacent tissues through immunohistochemical method. RT-PCR was used to detect MRP3 mRNA in 7 tumor cell lines and human embryo kidney cell lines 293T. Monoclonal antibody against MRP3 was employed to detect MRP3 protein expression through flow cytometry. Results There was significant difference for MRP3 expression in nucleus between the pancreatic cancer and normal tissues beside the cancer ( P= 0.003 ) while no significant difference was found in cytoplasm (P = 0. 472). MRP3 mRNA expression was found in 3 pancreatic carcinoma cell lines. The positive expression rate of MRP3 protein was relatively high in BxPC-3 and AsPC-1 cells (68. 5% and 33.6% respectively) while it was only 5.4% in PNAC-1. Conclusions The difference of MRP3 expression in pancreatic cancer and normal tissues lies mainly in nucleus. BxPC-3 may serve as cellular models for in vitro studies on multidrug resistance of pancreatic carcinoma.

Objective To explore the mechanism of melanoma differentiation associated gene-7(mda-7) in combination with adriamycin(ADM) killing the HCC HepG2 cells and reversing their multidrug resistance (MDR). Methods The experiment was conducted in three groups including the combined group, ADM group and mda-7 group. MTT assay and FCM were used to determine the differences among the 3 groups and clarify the reversing effect of combined treatment on multidrug resistance of the tumor cells. Expression levels of MDR-1, STAT-3, BCL-2, BAXmRNA were determined with real-time PCR. Western blotting was performed to observe the changes of proteins gp-l70, stat3,P-stat3, PKB, bcl-2,bax in all 3 groups. Result After transfection with 100VP/cell Ad. mda-7,the growth suppression rate of HepG2 treated by ADM (1.5 mg/L) rose from 17.46% to 79. 5%.According to the changes, killed HepG2 cells were increased by a factor of 4.55. times. MDR-1 mRNA was decreased from (16.49 ± 0. 11) to (5.48±0.05) and STAT-3 mRNA increased from (13.17±0. 08) to (21. 57±0. 11)(P＜0.05). Western blotting also showed that P-170 and PKB was decreased and the phosphorylation-stat-3 increased after the combined treatment. Conclusion Ad.mda-7 can reverse the multidrug resistance HepG2 cells. It inhibits the expression of MDR-1 mRNA,then arrests PKB protein and the signaling pathway of active stat-3 to induce apoptosis of HCC cells.

OBJECTIVE: To identify the newborns who should receive hearing evaluation by hearing screening in high risk newborns; to find and confirm the high risk factors of hearing disorders in high risk newborns. METHOD: The first screening was performed by DPOAE. Newborns did not passed the first screening undertook second screening using DPOAE + ABR. and newborns did not passed the second screening received hearing evaluation. High risk factors of hearing loss were found by Logistic regression analysis. RESULT: Three hundred and twenty-seven cases were screened. The positive ratio in first screening was 37.0%. The positive ratio in second screening was 11.0%. Ten cases were diagnosed as hearing loss and the incidence of hearing loss was 3.39%. High risk factors of hearing loss were asphyxiation, very low born weight (<1,500 g) and head and neck abnormality. CONCLUSION (1) DPOAE combined with ABR is credible and feasible in hearing screening of high risk newborns. (2) High risk factors of hearing loss were asphyxiation, very low born weight (<1,500 g) and head and neck abnormality in this study.
Female ; Hearing Disorders ; diagnosis ; epidemiology ; prevention & control ; Hearing Loss ; diagnosis ; epidemiology ; prevention & control ; Hearing Tests ; Humans ; Incidence ; Infant, Newborn ; Male ; Neonatal Screening ; Risk Factors