Gentamicin production at home and abroad was fermentation cycle long,production rate low,production costs high and short of competitiveness.Added a little quantity of lysine,glycine,tyrosine,methionine into the medium of gentamicin producing Micromonospora purpurea during inoculation could improve the metabolic capability of microbe,shorten fermentation cycle and increase production rate of gentamicin.The new method was successfully developed in shaking flask test and applied to 5L glass fermentor for one month.Average data of one month showed that the fermentation time per cycle was shortened about 40% and the yield of gentamicin per fermentor increased about 14% in comparison with the conventional technique.The production rate of gentamicin was increased 30% ~95%(according to the capability of strain).The quality of the end product gentamicin conformed to CP2000,BP2000 and USP26 pharmacopoeia.Therefore,the new method may reduce production costs of gentamicin remarkably,it shows strong competitiveness in the marketplace.

Objective To evaluate the expression and relationship of miR-100 and FZD-8, one of the major compo?nents of Wnt signaling pathway, and the correlation of their expressions with lymph node metastasis in patients with breast cancer. Methods The expression of miR-100 was determined in 50 samples of human breast cancer tissues and adjacent normal tissues by in situ hybridization. The correlation of miR-100 expression with lymph node metastasis was analyzed by Mann-Whitney U test. The expression of FZD-8 was measured in 50 samples of human breast cancer tissues and adjacent normal tissues by immunohistochemistry. The correlation of the miR-100 expression with the protein expression of FZD-8 was evaluated by Pearson rank analysis. Results The expression of miR-100 was significantly lower in human breast can?cer tissues than that in adjacent normal breast tissues [2.00 (1.00, 3.00) vs. 6.00 (3.50, 8.00)]. The miR-100 expression was lower in patients with lymph node metastasis than that in patients without lymph node metastasis [1.50 (1.00, 2.75) vs. 3.00 (2.00, 4.00)]. The expression of FZD-8 was significantly higher in human breast cancer tissues than that in adjacent normal breast tissues [8.00 (6.00, 9.00) vs. 6.00 (3.75, 9.00)]. The miR-100 expression was negatively correlated with the FZD-8 pro?tein expression in human breast cancer tissues (rs=-0.592, P<0.001). Conclusion The miR-100, as an anti-metastasis-miRNA, may involve in the metastasis of breast cancer, which may be related with the regulation of the expression of FZD-8.

OTX1 gene is one of the pivotal transcriptional factors involved in the neurogenesis.In order to overexpress the OTX1 gene in distinct cell types and find out its contribution to the proliferation and differentiation of stem cells in vitro,OTX1 cDNA was subcloned into lentiviral vectors.The resulting constructions pDUETOTX1,pDUETGFPOTX1 and pDUETGFP were packaged in 293 cells producing viral particles to transduce 293T cells,SY5Y cells,mouse embryonic stem cells and E15 neural stem cells.It was proved that the transferred OTX1 gene was located in the nuclei of the transduced cells in stead of plasma.Lentivirus is an ideal vector delivering gene to different cells.The overexpression of OTX1 in transduced 293T cells were validated by Western blot and immunofluorescence.

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

OBJECTIVE To investigate the anti- tremor effect and mechanism of baicalein on oxotremorine- induced muscle tremor in mice. METHODS The acute model of muscular tremor was induced by intraperitoneal injection of oxotremorine, and the latency, duration and frequency of muscle tremor in mice were measured immediately; the saliva of mice was measured to reflect the correlation between tremor and peripheral nerve function; the aim of this study was to determine the content of MDA and the activity of GSH-PX, and to investigate the anti-oxidation of mice with tremor model. The activity of acetylcholinesterase (AchE) and acetylcholine transferase (ChAT) can indirectly reflect the level of acetylcholine in the brain. The level of monoamine neurotransmitters in brain tissue was determined by high performance liquid chromatography (HPLC-ECD). RESULTS The animals in the model group appeared obvious tremoring, salivating and erecting and other symptoms. Compared to the model group, there was no obvious inhibitory effect on the administration of each dose. After 7, 14, 21 and 28 d of continuous administration, the latency, duration and tremor frequency of tremor mice were significantly shortened, the levels of acetylcholine were significantly decreased, the changes of DOPAC and DA neurotransmitters in the brain of model group were recovered, regulate the dynamic balance of acetylcholine and dopamine in the brain. CONCLUSION Long- term administration can improve the tremor behavior of mice, the mechanismmay be related to the regulation of neurotransmittersin brain.

Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78％ ± 0.52％ vs.0.47％ ± 0.15％,P＜ 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P＜ 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P＜ 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P＜ 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P＜ 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P ＜ 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P ＜ 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P ＜ 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .

Objective: To explore the expression of hypoxia inducible factor-1α (HIF-1~) and the correlation between HIF-1α and apoptosis after traumatic brain injury.Methods: Using experimental traumatic brain injury in the rats, the expression of HIF-1α was studied by immunohisto-chemistry in cerebral tissue, apoptotic cell death was evaluated with TUNEL (transferase-mediated XdUTP nick end labeling ), and double-labeled immunohistochemistry and TUNEL methods were used to investigate the relationship between HIF-1α and apoptosis.Results: There was remarkable difference in the expression of HIF-1α between the experimental groups and the control groups (P ＜ 0.01), in the experimental groups,the expression of HIF-1α at 48 hours was highest; the evidence of apoptotic cell death after experimental traumatic brain injury was found by TUNEL; the apoptotic percentage increased or decreased according to the changes of the positive expression of HIF-1α (r = 0.99).Conclusions: The results suggest that secondary brain ischemia plays a crucial role in apoptotic cell death after traumatic brain injury; HIF-1α can prompt apoptotic cell death after experimental traumatic brain injury.

16 and the distribution of severe trauma more than 2 anatomic parts.They were randomly divided into two groups:intensive insulin treatment group(n=31)and control group(n=31).Intensive insulin treatment group received insulin with insulin pump in order to maintain blood glucose levels at 4.0-6.1 mol/L,while the control group received routine insulin treatment in order to mmaintain blood glucose levels at 10.0- 11.0 mol/L.Plasma levels of TNF-?,IL-1,IL-6, CRP,APACHEⅡscores and cure rate were analyzed before and after the treatment.Data was expressed as mean?standard deviation.Two- tailed T test and ANOVA were used for comparison in SPSS 10.0,and changes were considered as statistically significant if P value was less than 0.05.Results After the intensive insulin treatment, patient's hemodynamic parameter apparently improved,APACHEⅡscores descended,and the levels of TNF-?, Ib-1,IL-6,CRP all declined,in comparison with control group,there were significant differences. Intensive insulin treatment might improve patient's general condition and decrease complications and mortality of severe multiple trauma.

Objective:To investigate the feasibility of real-time screening and diagnosing fundus diseases with Wild Field Imaging System (WFIS SW-8000) in the eyes with dense cataract patients.Methods:A series of case-observational study was carried out.Ninety-six dense cataractous eyes of 90 patients with suspected fundus diseases were included in Affiliated Hospital of Nantong University from April to July 2019.Lens phacoemulsification combined with intraocular lens implantation was performed after the opacity lens was removed, and fundus examination was performed with WFIS SW-8000.Super field lens, optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) was used to verify the fundus findings after surgery, which were compared with WFIS SW-8000.This study protocol was approved by Ethics Committee of Affiliated Hospital of Nantong University and followed the Declaration of Helsinki.Written informed consent was obtained from each patient prior to any medical examination.Results:Among 96 eyes, fundus diseases were detected in 40 eyes with the detection rate of 41.67% by WFIS SW-8000 examination, including dry age-related macular degeneration (AMD) in 2 eyes, wet AMD in 3 eyes, polypoidal choroidal vasculopathy (PCV) in 1 eye, high myopia retinopathy (HMR) in 7 eyes, central retinal vein occlusion (CRVO) with macular edema (ME) in 1 eye, mild non-proliferative diabetic retinopathy (NPDR) in 12 eyes, moderate NPDR in 7 eyes, severe NPDR in 4 eyes and proliferative diabetic retinopathy (PDR) in 3 eyes.Fundus diseases were detected in 45 eyes with detection rate of 46.88% after surgery by a super field lens, OCT and FFA.The detectable rate of digital retinal camera examination and super field lens, OCT and FFA showed no significant difference between intraoperation and postoperation (χ 2=0.528, P=0.468). Twenty-two eyes with fundus neovascular disease or macular edema requiring drug intervention were identified intraoperatively, and intravitreal anti-inflammatory and/or anti-vascular endothelial growth factor (VEGF) drugs were injected in 11 eyes during the operation. Conclusions:WFIS SW-8000 is a useful tool for the accurate and convenient method in real-time fundus examination during phacoemulsification, which is feasible and helpful for timely intervention in and treatment of fundus diseases.