Pullulan is a linear glucosic polysaccharide produced by the polymorphic fungus Aureobasidium Pullulans, which has long been applied for various applications in medical and food industry due to its security, stability and low adhesive ability.At present, the two problems in restricting pullulan industrial production are the low polysaccharide production and melanin secreted which is hard to erase completely, giving the following process some problem.As a starting point, this review article collects and analyzes the progress on the breeding of pullulan high-yield strain without melanin in recent years, in order to find more efficient strains breeding methods, laying a foundation for further breeding of pullulan high-yield strain without melanin.

AIM To explore the radioprotective function of Xihuang Capsules (Bovis Calculus artijactus,Moschus,Olibanum,Myrrha) in patients with acute radiation-induced oral mucositis and its mechanisms.METHODS Eighty patients with nasopharyngeal carcinoma undergone radiotherapy were randomized into two groups:treatment group (radiation plus Xihuang Capsules) and control group (radiation alone).The comparison of two groups in the onset and the tolerated dose of mucositis was made according to the acute radiation injury classification standard.The levels of TNF-α and IL-6 in saliva were detected by ELISA.RESULTS After the treatment with Xihuang Capsules,the onset of oral mucositis delayed significantly and the tolerated dose elevated markedly comparing with the control group (t =2.180,12.930,P ＜ 0.05).The morbidity rate of Ⅲ-N-degree of oral mucositis in the treatment group was lower than that of the control group at the dosage of 40 Gy and 70 Gy,respectively (Z =3.661,4.270,P ＜0.01).Furthermore,there was no difference in the levels of TNF-α and IL-6 in the pre-treatment in the two groups (t =1.010,1.469,P ＞ 0.05).With the increase in radiant dose,the levels of TNF-α and IL-6 in the two groups both elevated dramatically,and peaked at the dose of 40 Gy,but both levels in the treatment group were lower than those of the control group (t =8.305,6.069,P ＜ 0.05).When DT =70 Gy,the levels of TNF-α and IL-6 were lower than pre-radiation,and the levels of TNF-α and IL-6 in the treatment group were lower than those in the radiation alone group (t =3.835,2.488,P ＜ 0.05).CONCLUSION Xihuang Capsules can delay and reduce acute radiation-induced oral mucositis and improve radiation tolerated dose,so it may involve the release of TNF-oα and IL-6 in saliva.

Objective To study the risk factors and pathogenesis of pulmonary infection in acute stroke patients. Methods A retrospective analysis was taken in 257 acute stroke cases to investigate the incidence of pulmonary infection and its relationship with the underlying disease, disturbance of consciousness, invasive operation,the type of pathogenic bacteria and drug susceptibility. Results The positive rate of pulmonary infection in acute stroke patients was high and related to many factors including severity of stroke, disturbance of consciousness,primary diseases, and invasive examination etc. The most popular infective bacteria were Gram-negative bacteria. The imipenem was most sensitive to Gram-negative and Gram-positive bacteria, followed by Amikacin and piperacillin too. Conclusions The pulmonary infection in cerebral stroke patients is caused by many factors, of which Gramnegative bacteria play the most important role. Identification of the risk factor and pathogenic bacteria is helpful in treating cerebral stroke infectious complications.

AIM: To study the effects of disulfiram/copper complex (DSF/Cu) on ultrastructures and mechan-ical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy (AFM) based on the nanoscale resolution and piconewton force measurement level.METHODS: The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10A cells induced by DSF/Cu were compared by flow cytometry.The cell surface morphology, ultra-structure, height, width and roughness were detected by AFM.The effects of DSF/Cu on the hardness (Young’s modu-lus) of the 2 kinds of cells were determined by AFM with indentation technique.RESULTS: DSF/Cu significantly in-duced apoptosis of the MCF-7 cells in a dose-dependent manner, whereas had little effect on the MCF-10A cells.The cell cycle analysis showed that DSF/Cu induced G2 /M arrest in the MCF-7 cells, but led to G0 /G1 arrest in the MCF-10A cells.The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology, and the filopo-dia were retracted obviously, even some became into lamellipodia, or disappeared completely after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.The quantitative analysis indicated that the MCF-7 cells showed smaller width and
larger height, and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.However, little effect in the MCF-10A cells was observed.The biomechanics test at a single cell level demonstrated that the Young’s modulus of the MCF-7 cells and MCF-10A cells were both increased, yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10A cells after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.CONCLUSION: DSF/Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.

Objective To analysis the difference of lymph node cleaning and operative complication rate between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer.Methods A summary of 62 patients with thoracic segment esophageal cancer in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from August 2012 to August 2014,who were carried with thoracoscope surgery,were randomly chosed and designed as the thoracosc0Pe group,and 62 patients with thoracic segment esophageal cancer over the same period,carried out with routine thoracotomy,were designed as the control group.All the clinical data of the two groups were collected.The total of thoracic lymph node cleaned and the group of thoracic lymph node cleaned were compared between the two groups.The operation time,intraoperative blood loss,chest tube placement time and postoperative hospital duration were collected and compared.All the patients were followed up at least for one year.The incidence of postoperative complications such as pulmonary infection,pneumothorax,atelectasis,recurrent laryngeal nerve injury and anastomotic leakage in the follow-up period were compared.The follow-up time,mortality and recurrence rate were compared.Results The total of thoracic lymph node cleaned(13.36±3.28) and the group of thoracic lymph node cleaned(3.35±0.84) in the thoracoscope group were lower then these of the control group ((14.22± 2.78) and (3.58±0.75)),but with no statistical difference (t =1.57,1.61,P＞ 0.05).The operation time of the thoracoscope group((314.63±38.72) min) were higher then that of the control((217.46±41.54) min),and the intraoperative blood loss ((205.73 ± 114.38) ml),chest tube placement time ((6.83 ± 1.92) d) and postoperative hospital duration((18.47±5.36) d) of the thoracoscope group were remarkably lower then these of the control ((345.72 ±175.62) m1,(10.04±2.41) d,(22.65±6.84) d,t=13.47,5.26,8.20,3.79,P＜0.05).The incidence of pulmonary infection (4.8％ (3/62)) and atelectasis (1.6％ (1/62)) of the thoracoscope group,were evidently lower then these of the control (17.7％ (11/62),1.3％ (7/62),x2 =5.15,4.81,P＜0.05).There was no significant difference in mortality and recurrence ratebetween the two groups during the follow-up period (3.2％ (2/62) vs.8.1％ (5/62),11.3％ (7/62) vs.14.5％ (9/62),x2 =1.36,0.29,P＞0.05).Conclusion There are no significant difference inlymph node cleaning between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer,but thoracoscope surgery can shorten the length of hospital duration,reduce the intraoperative blood loss,chest tube placement time and postoperative complications.So the thoracoscope surgery is a safe and feasible operation for patients with thoracic segment esophageal cancer.

ACKGROUND:Autism is a kind of neurobiological disease.The increase of plasma concentration of 5 hydroxytryptamine(5 HT) is considered to be one of study results,which is the most definite and has the best repetition,for the biological investigation of autism. Whether the season of birth and maternal mood during pregnancy are correlated with the occurrence of autism? OBJECTIVE:To compare the differences of the plasma concentration of 5 HT,season of birth and the maternal mood during pregnancy between autistic and normal children,and probe into the cause of increased 5 HT in autism. DESIGN:An observational comparative study taking autistic and normal children as the subjects. SETTINGS:A clinical psychological department of a university hospital;Mental Health Institute of the Xiangya Second Hospital;A rehabilitation center for dumb children of city grade. PARTICIPANTS:Children in the autistic group(n=54),who were in accordance with the diagnostic criteria of autism in Diagnostic and Statistical Manual of Mental Disorders Fourth Edition(DSM IV) and had the total score of social adaptive ability scale(SAAS) over 40, were selected from the Outpatient Clinic for Children of the Mental Health Institute,Xiangya Second Hospital of Central South University and the training class of autism of Shenzhen Special Education School;Children in the normal control group,who had no obvious somatic and neurological diseases,and developing delay detected by physical,neurological and mental status examinations, rough detection of developing level,were selected from the primary school and kindergarten in Changsha and Shenzhen cities. INTERVENTIONS:The plasma level of 5 HT was examined, season of birth and maternal mood during pregnancy were investigated in autistic and normal children.The children with autism were accessed with SAAS. MAIN OUTCOME MEASURES:The plasma level of 5 HT,the season of birth and the maternal mood during pregnancy in both groups. RESULTS:The score of SAAS in children with autism was 67.65± 1.67.The plasma level of 5 HT was(0.78± 0.32) μ mol/L in the autistic group,which was significantly higher than that in the normal control group [(0.62± 0.16) μ mol/L](t=3.17,P=0.002). There were 17 children with autism had increased 5 HT.Compare with normal children,most children with autism were born in winter and spring,and their mother had obvious mood problems during pregnancy. CONCLUSION:The 5 HT level is higher in children with autism than in normal children.Most children with autism were born in winter and spring,and their mother had obvious emotional problems during pregnancy.There are phase of high functioning of 5 HT in mother in key period of fetus cerebra development that may be one of the causes of hyperserotonemia in autism,and that is possibly one of the etiologies of autism.

Objective To explore the measurement of(1,3)-β-D glucan in plasma for the diagnosis of pulmonary fungal infections in pulmonary tuberculosis patients. Methods 40 pulmonary tuberculosis patients with pulmonary fungal infections in Guangzhou chest hospital from January 2015 to December 2015 were enrolled as a test group,among which 35 were confirmed and 5 were suspected pulmonary fungal infections. 52 pulmonary tuber-culosis patients without fungal infections were selected as a control group.(1,3)-β-D glucan content(G test)in this 92 patients plasma were detected. The results of G tests were compared with those from etiological diagnosis to assess the performance of G test. Results 13 strains of candida albicans,13 strains of aspergillus,2 strains of candida tropicalis,2 strains of candida glabrata and 6 strains of other yeast were obtained from patients of test group,but no fungal identified from those of control group. The median of G test in test group and in control group was 126.1 and 29.56 pg/mL,respectively,the level in test group was significantly higher than that in control group (P<0.001). 35 cases were identified as positive and 5 were negative in test group by G test ,while 41 cases were identified as negative and 11 were positive in control group. The sensitivity,specificity,positive predictive value, negative predictive value ,concordance and Youden index of G test were 87.5%,78.85%,76.09%,89.13%, 82.6%and 0.663,respectively. Conclusions Candida albicans and aspergillus are more common pathogens than the other fungi isolated from pulmonary tuberculosis patients with pulmonary fungal infection. G test ,used in pul-monary tuberculosis with pulmonary fungal infections diagnosis,is reliable and fast,and has a higher sensitivity, specificity and accuracy.

Aim To study the mechanism of DSF-Cu induced apoptosis of human nasopharyngeal carcinoma CNE-2 Z cells by affecting the function of mitochondria and cytoskeleton. Methods The cell cycle,the rate of apotosis,the levels of intracellular ROS and MMP in CNE-2 Z cells were tested by flow cytometry after trea-ted with different concentration of DSF-Cu. The chan-ges of the cell surface morphology, ultrastructure, cell height, width and roughness were detected by AFM. The distribution and reorganization of cytoskeleton F-actin were observed by Laser scanning confocal micro-scope. Results Cells were incubated with different concentration of DSF-Cu ( 0 ~200 nmol · L-1 ) for 24 h, the apoptotic ratio increased significantly and the treatment of DSF-Cu resulted in a concentration-de-pendent accumulation of CNE-2Z cells in G2/M phase. Furthermore,the treatment of DSF-Cu was able to in-crease the production of intracellular ROS and decrease the MMP in CNE-2Z cells. In addition,AFM imaging showed that compared to the control group,with the in-crease of DSF-Cu concentration,the CNE-2Z cells be-came smaller, cytoplasm condensed, the height in-creased,and the surface roughness reduced. Moreover, the filopodia became shorter, shrinked and even com-pletely destroyed after treated with different concentra-tion of DSF-Cu. At last,the LSCM image showed that the fluorescence intensity of F-actin networks was de-creased, then the structure was rearranged and de-stroyed obviously by treated with DSF-Cu. Conclusion DSF-Cu can induce apoptosis and arrest cell cycle at G2/M phase in CNE-2Z cell through a mitochondria-dependent pathway. Above findings highlight the appli-cations of AFM at the single cell level for the investiga-tion of antineoplastic drug in nasopharyngeal carcinoma therapy.

Aim To explore the role of heat shock protein 90(Hsp90)in hydrogen sulfide-induced protection against PC12 cells injuries elicited by cobalt chloride(CoCl_2).Methods Hydrogen sulfide preconditioning against CoCl_2-induced injury model was set up in PC12 cells.Cell viability was tested by using cell counter kit-8.Morphological changes in apoptotic PC12 cells were detected by Hoechst 33258 staining and photofluorography.Apoptotic rate was evaluated by propidium iodide(PI)staining and flow cytometry(FCM).The expression of Hsp90 was evaluated by Western blot.Results Hsp90 expression was upregulated after treatment with 400 μmol·L~(-1) sodium hydrosulfide(a H_2S donor),peaking at 3 h,returning to the basal level at 24 h.Furthermore,H_2S preconditioning obviously enhanced the upregulation of Hsp90 expression induced by CoCl_2.H_2S preconditioning markedly protected PC12 cells against injuries induced by CoCl_2,increasing the viability of cells and decreasing the percentage of apoptotic cells.17-allylamino-17-demethoxygeldanamycin(17-AAG),an inhibitor of Hsp90,antagonized H_2S preconditioning-induced Hsp90 activation and the adaptive cytoprotection.Conclusion H_2S can protect PC12 cells against CoCl_2-induced injuries,and upregulating the expression of Hsp90 may be one of the mechanisms underlying cytoprotection induced by H_2S preconditioning.

AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.