OBJECTIVE: To investigate the optimizing conditions in isolation of the side population in laryngeal carcinoma cell line Hep-2. METHOD: Single-cell suspension cells were detached from the culture flask with trypsin EDTA, at a concentration of 1 x 10(6) cells/ml. (1) The trail Samples were incubated with Hoechst33342 at a concentration of 5 microg/ml, 9 microg/ml, 10 microg/ml, 11 microg/ml for 90 minutes. (2) They were incubated with Hoechst for 50, 70, 90, 110, 130 min in water bath individually. (3) The single-cell suspension were incubated Hoechst in water bath and in thermostat each. (4) The two different density of cells were harvested, which were 100% and 70%, and then di gest into single-cell suspension. Once incubation finished, suspended in phosphate buffered saline (PBS), then test SP% by flow cytometry. Among all groups,Verapamil hydrochloride was added to the control samples, incubated at 37 degrees C for 30 minutes, the other condition were keep the same with their trial groups. RESULT: (1) The percentage of Hoechst-negative cells in trial group was (39.96 +/- 0.24)%, (26.23 +/- 0.39)%. (18.79 +/- 0.02)%, (19.01 +/- 0.14)% at the concentration of 5 microg/ml, 9 microg/ml, 10 microg/ml, 11 microg/ml respectively, when the PI-positive cells were (30.45 +/- 0.63)%, (49.9 +/- 0.42)%, (50.12 +/- 0.68)%, (64.16 +/- 0.39)% separately. (2) Varying the duration of staining incubation showed that there was a typical FACS pattern and SP% was constant when the incubation was at least 90 min. (3) Compare to water bath, SP% was more than in thermostat, the SP% was (18.67 +/- 0.45)%, (22.6 +/- 0.50)% respectively; (4) Cell density is also responsible for SP%. The low density the cell is, the less in SP%. SPSS13.0 was used in statistical analysis, the groups were compared using t-Test. P < 0.05 was considered statistically significant. CONCLUSION The optimum concentration and duration of incubation of Hoechst33342 in isolation of the side population cells in laryngeal carcinoma cell line Hep-2 is 10 microg/ml and 90 min. Incubated in water bath is better than in thermostat. The best staining cell density is around 80%-90%.
Cell Count ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; pathology ; Neoplastic Stem Cells ; cytology ; Side-Population Cells ; cytology

Apathy is a common neuropsychiatric complication after stroke,the identification of apathy after stroke and its related factors and providing an effective intervention on it may contribute to the rehabilitation in patients with stroke.This article reviews the incidence of apathy after stroke,pathophysiological mechanisms,diagnostic evaluation and treatment.

Objective To explore a novel surgical treatment for chronic suppurative otitis media (CSOM) and evaluate its treatment effect.Methods All 97 patients with chronic suppurative otitis media were chosen to be treated using this new surgical method.The skin of the external auditory canal was maintained intact.Open radical mastoidectomy was used to complete clean-up lesions ; the fascia of pedicled temporalis myofascia (PTM) was used to repair the tympanic membrane.The pedicled temporalis fascia,pedicled postauricular periosteal flap and intact skin of the external auditory canal were used in reconstruction of the posterior wall of external auditory canal.Pure tone audiometry (PTA) was performed before and after surgery,recording the air conduction and bone conduction thresholds at 0.5 kHz,1.0 kHz,2.0 kHz,4.0 kHz.The average of the patient's air and bone conduction hearing thresholds was recorded at the 4 frequencies.External auditory canal gauze was removed 3 weeks after surgery.All subjects were followed up for over 2 years.Comparison of hearing thresholds (PTA) was made ① Before and 4 weeks after surgery.② Before and 2 years after surgery.Hearing function comparison include air conduction (AC),bone conduction (BC) and air-bone gap (ABG) analysis.SPSS 16.0 was used in statistical analysis.Pre-and postoperated AC,BC and ABG were compared using T-test.P ＜ 0.05 was considered statistically significant.Results The healing rate of post-operated tympanic membrane was 95.88％ (93/97).Ninty-six ears had 2-year follow-up,and 1 patient was lost in follow-up.There were 2 patients presented with eardrum perforation during the follow-up,and the 2-year healing rate of tympanic membrane perforation was also 93.85％ (92/97).In 96 ears with 2-year followed-up,the average of pre-AC was (52.10 ±3.96) dB,the average of post-AC was (35.67 ±2.52) dB; the average of preABG was (36.6 ± 5.2) dB,and the average of post-ABG (± SD) was (12.14 ± 6.20) dB.Statistical analysis showed significant difference between preoperative and postoperative AC or ABG values (P ＜ 0.05).Conclusion The present surgical procedure broke through the existing conventional mastoidectomy of making a surgical incision in the posterior wall of the external auditory canal.This procedure cleared the lesion completely and preserved the physiological function of the external auditory canal.The acoustic systems and state of gasification to the mastoid tymnpanum were reconstructed,rehabilitated and maintained.The healing rate of hearing and tympanic membrane perforation was improved.

Objective:To experimental study the inhibiting effects of SBHL on the growth of cervical cancer cells and to investigate the mechanisms.Methods:The proliferation of ME180 cells was observed by MTT assasy; The changes of cells cycles and apoptosis of ME180 cells treated with SBHL were analyzed by agar gel electrophoresis and FCM assay;. The changes of the ultra structure of cells was observed by Transmission eleconmicrograph.Results:The proliferation of ME180 cells was inhibited after treating on SBHL for 24 h.The inhibiting effect increased granually following the concentration of SBHL(P

OBJECTIVETo observe the difference in clinical efficacy on focal vitiligo treated with heat-sensitive moxibustion in comparison with medication, and discuss its effect mechanism.

METHODSSixty-eight cases were randomized into a moxibustion group (38 cases) and a medication group (30 cases). Additionally, 20 healthy persons were selected randomly as a normal group. In the moxibustion group, the heat-sensitive moxibustion was applied to Hegu(LI 4), Quchi(LI 11), Yanglingquan(GB 34), Zusanli(ST 36), Xuehai(SP 10) and the others, once a day. In the medication group, triamcinolone acetonide cream was used externally and locally, twice a day. In the two groups, the treatment of 15 days made one session. The efficacy was observed after continuous treatment for 3 sessions. The hemorheology test was done in all of the subjects. The radioimmunoassay was adopted to determine the levels of Interleukin 2 (IL-2), Interleukin 6 (IL-6), Interleukin 10 (IL-10) and tumor necrosis factor (TNF-alpha) before and after treatment.

RESULTSThe levels of IL-6, IL-10 and TNF-alpha in vitiligo patients were higher significantly than those in the normal group (P<0. 01, P<0. 05), the level of IL-2 was lower significantly than that in the normal group (P<0. 01) before treatment. After 3 sessions treatment, IL-2 level was increased significantly in the moxibustion group and the levels of IL-6, IL-10 and TNF-alpha were reduced, without significant differences as compared with the normal group (all P>0. 05). But the differences were significant as compared with those in the medication group (all P<0. 05). The curative and remarkably effective rate was 76. 3% (29/38) after treatment in the moxibustion group, which was higher significantly than 13. 3% (4/30, P<0. 05) in the medication group.

CONCLUSIONHeat-sensitive moxibustion achieves very good clinical efficacy on focal vitiligo, which is probably via promoting blood circulation and regulating the levels of IL-6, IL-10 and TNF-alpha.

Adolescent ; Adult ; Child ; Female ; Hot Temperature ; Humans ; Male ; Middle Aged ; Moxibustion ; Triamcinolone Acetonide ; administration & dosage ; Tumor Necrosis Factor-alpha ; Vitiligo ; drug therapy ; therapy ; Young Adult

Abdominal Muscles ; physiology ; Abdominal Pain ; diagnosis ; physiopathology ; Animals ; Animals, Newborn ; Colon ; innervation ; Electrodes ; Electromyography ; Female ; Pain Measurement ; methods ; Pain Threshold ; physiology ; Rats ; Rats, Sprague-Dawley ; Viscera ; Visceral Afferents ; physiology

Objective To investigate the feasibility and efficacy of combined preoperative chemora- diation program followed by radical surgery in bulky early or moderate uterine cervical cancer.To determine the incidence and predictive value of rasidual disease in the radical hysterectomy specimens after CCR. Methods Thirty-five patients with uterine cervix carcinoma from January 2001 to September 2003 were treated with preoperative external beam radiotherapy to 3060 cGy in 4 weeks.Patients received concurrent continuous infusion cisplatin(20 mg/m~2)on day 1,8,15,22 of four weeks and 5-Fu(4 g)chemotherapy during the first four days and the last four days of the radiation course.Radical surgery was not performed until 4 weeks after the completion of the preoperative treatment.Results Clinical response was 85.71%,complete response,54.29 %;partial response,31.43 %.The analysis of the surgical specimens showed 19 patients had revealed residual disease and 16 patients had revealed no residual disease,the complete pathological response hed been 45.71%.Three-year survival rate was 87.14 % in 35 patients.Three-year survival rate of residual disease patients(76.32 %)was significantly lower than that of no residual disease(100.0 %)(P=0.0358). Three-year survival of cervical stromal invasion less than 1/2 and invasion extra stromal was 100.0 %, 41.67 % respectively(P=0.0109);three-year survival of lymph-vascular space involvement and no lymph- vascula space involvement was 0,85.29 %,respectively(P=0.0148).Conclusion Combined preoperative chemoradiation program followed by radical surgery in bulky early or moderate cervical cancer could reveal an effective efficacy with a tolerant complication.Residual disease is an independent and strong predictive factor.

Objective To evaluate the diagnostic and prognostic value of the serum procalcitonin ( PCT ) level in the non-acquired immune deficiency syndrome ( AIDS ) immunocompromised critically ill patients suspected to have infection. Methods A retrospective study was conducted in the non-AIDS immunocompromised patients who were admitted to Department of Critical Care Medicine of Xiangya Hospital, Central South University during January 2011 to December 2014. Demographic characteristics, underlying disease, acute physiology and chronic health evaluationⅡ( APACHEⅡ) score at admission, and clinical records including baseline and peak levels of temperature, white blood count ( WBC ), PCT, and survival rate within 28 days, infection focus, infectious agents ( bacterial, fungi or mixed infection ), and the severity of infection ( sepsis, severe sepsis, or septic shock ) were recorded. Receiver operating characteristic ( ROC ) curve was plotted, and the diagnostic and protective value of above parameters was evaluated. Results A total of 98 patients ( 43 male and 55 female ) were enrolled in the study with a median age of 44 ( 28, 52 ) years old and a median APACHEⅡscore of 17 ( 11, 20 );47 with malignant hematological tumor, 45 with autoimmune diseases, and 6 post solid organ transplantation. Among them 53 patients ( 54.1%) died within 28 days. Twenty-seven patients were diagnosed as systemic inflammatory response syndrome ( SIRS ) without infection. Among 71 patients with infection, 45 were diagnosed as bacterial infection, 10 with fungal infection, and 16 with mixed infection. Sepsis was diagnosed in 7 patients, severe sepsis in 32 patients , and septic shock in 32 patients .①There was no statistical significance in the baseline and peak levels of PCT and WBC, or baseline level of temperature between the groups of SIRS patients without infection and infected patients. The peak level of temperature was significantly higher in the patients with infection as compared with that of the SIRS without infection patients [℃:39.4 ( 38.9, 40.0 ) vs. 38.8 ( 37.8, 39.2 ), Z=-3.268, P=0.001 ]. It was showed by subgroup analysis that in patients with hematological malignant disease or autoimmune diseases, higher level of body temperature was found in infection group compared with non-infection SIRS group [℃:39.5 ( 39.0, 40.0 ) vs. 39.0 ( 38.4, 39.4 ), Z=-2.349, P=0.019;39.0 ( 38.4, 39.5 ) vs. 38.2 ( 37.0, 38.9 ), Z=-2.221, P=0.026 ].②The baseline level of PCT (μg/L ) were 0.54 ( 0.20, 4.19 ), 2.78 ( 0.50, 9.54 ), 1.00 ( 0.45, 6.89 ), and 0.22 ( 0.07, 1.86 ) in non-infection SIRS patients or the patients with bacterial, fungal, and mixed infection, respectively. The peak level of PCT (μg/L ) were 4.19 ( 1.95, 13.42 ), 12.37 ( 3.82, 45.89 ), 1.82 ( 0.49, 17.86 ), and 5.14 ( 2.66, 12.62 ), respectively, in each subgroup. When the comparison was conducted among the patients with different infectious agent, the baseline level of PCT in patients with bacterial infection was significantly higher than that in SIRS patients without infection ( P=0.026 ) and mixed infection patients ( P=0.001 ), and the peak level of PCT was significantly higher than that in the SIRS patients without infection ( P=0.009 ) and the patients with fungal infection ( P=0.016 ). ROC curve showed that the higher value was found in the baseline and peak levels of PCT for diagnosis of septic shock in all patients [ area under ROC curve ( AUC ) of baseline level = 0.681±0.054, P = 0.001; AUC of peak level = 0.690±0.054, P=0.002 ], and the same value was also found in the baseline and peak levels of PCT for diagnosis of bacterial infection in the patients with malignant hematological tumor ( AUC of baseline level=0.687±0.080, P=0.008;AUC of peak level=0.697±0.079, P=0.021 ).③The peak level of PCT (μg/L ) were 4.05 ( 0.53, 31.22 ), 5.78 ( 2.14, 16.68 ), and 11.64 ( 2.94, 58.14 ) in subgroup of patients with sepsis, severe sepsis and septic shock, respectively, and they showed no statistical significance among subgroups ( P>0.05 ). A high serum level of peak PCT strongly indicated the presence of septic shock ( AUC=0.646±0.060, P=0.019 ), especially in the subgroup of patients with systemic autoimmune disease ( AUC=0.689±0.081, P=0.035 ).④The peak level of PCT (μg/L ) in the APACHEⅡ>18 group ( 38 cases ) was significantly higher than that of APACHEⅡ≤18 group [ 60 cases, PCT (μg/L ):11.64 ( 3.36, 39.39 ) vs. 4.42 ( 1.32, 14.70 ), P=0.016 ];there was a certain correlation between the peak level of PCT and the severity of the disease.⑤The peak level of PCT in death group was significantly higher than that of the survival group [μg/L:9.07 ( 3.05, 33.09 ) vs. 4.19 ( 1.26, 14.61 ), P=0.043 ]. ROC curve showed that the peak level of PCT might be valuable in predicting the prognosis in immunocompromised patients ( AUC=0.619±0.057, P=0.043 ). Conclusions The serum level of PCT is found to be a reliable marker for the diagnosis of bacterial infection in immunocompromised critical patients, especially in those with hematologic malignancy. Additionally, PCT provides a useful tool for evaluating the severity of infection and the prognosis of critically ill patients.

Objective To summarize the experimence of nonmyeloablative allogeneic peripheral blood cell transplantation in the treatment of chronic leukemia. Methods Seven patients, including 6 cases of chronic myeloid leukemia (in chronic phase), one of chronic lymphoid leukemia (in third stage), with HLA-identical siblings donor received allogeneic peripheral blood stem cell transplantation after a nonmyeloablative conditioning. Results All of them were engrafted with donor cells (4 with full of donor cells grafted, 3 with mixed chimerism) and recovered hematopoiesis (WBC recovered to more than 0.5 ?10 9/L during postoperative 9 day to 21 day and platelet recovered to more than 30?10 9/L during postoperative 11 day to 28 day). One of them developed a GVHD of degree IV. One of them developed aGVHD of degree I. Conclusion This procedure is much safe, effective and of less complications than the myeloablative condioning regimens and may represent another new approach in the management of patients with chronic leukemia.

AIM:To evaluate the clinical efficacy and safety of balloon dilatation in infants with lacrimal passage obstruction.METHODS:Totally 86 patients (116 eyes) with lacrimal duct obstruction from July 1, 2015 to June 30, 2016 were randomly divided into two groups according to the digital table.The observation group (43 cases, 60 eyes) were operated with balloon dilatation and the control group (43 cases, 56 eyes) were treated with duct exploratory operation.The patients were followed up for 6mo to compare the efficacy.RESULTS:At the 6mo postoperatively, the primary cure rate and total cure rate in the observation group were significantly higher than those in the control group.There was significant difference between the two groups (P<0.05).CONCLUSION:Balloon dilatation operation is safe, and its clinical efficacy is better than lacrimal duct exploratory operation, is an effective way to treat lacrimal duct obstruction in infants.