A strain Actinobacillus succinogenes CGMCC 1593 was selected as the parent strain.After UV-EMS and UV-DES treatments respectively,seven mutated strains with subtle improvements in acid tol-erance were obtained,and were subjected for recursive protoplast fusion.Through three rounds of genome shuffling,four shuffled strains with both higher yield and acid tolerance were obtained.The shuffled strain namely F3-21 could even survive at pH 5.2.The comparison of the shuffled strains and the parent strain for succinic acid production was also studied here.After 48 h of shake-flask fermentation,the succinic acid concentration of F3-21 was 48% higher than that of the parent strain.When F3-21 was carried out in a 5 liter stirred bioreactor with pH controlled 5.6~6.0,the accumulation of succinic acid in 48 h fermentation attained 38.1 g/L,which was increased by 45% compared with that of the parent strain(26.2 g/L).While pH was controlled at 6.5~7.0,the production of succinic acid in 32 h fermentation attained 40.7 g/L.When F3-21 was carried out in fed-batch fermentation,succinic acid concentration of 67.4 g/L was reached in 72 h fer-mentation.These results indicated that the genome shuffling could improve the acid tolerance and the suc-cinic acid production of A.succinogenes CGMCC 1593.

Andrographolide total ester sulfonate (ATES) injection is one of the products of traditional Chinese medicine (TCM) currently used against viral infection in China.ATES injection was approved for manufacturing and marketing in January 2002.It is indicated for acute respiratory infections,tonsillitis,chronic obstructive pulmonary disease,influenza,foot and mouth disease,bronchiolitis,herpangina,mumps,infectious mononucleosis and psychosis.However,its usage also carries risk.We investigated the use of ATES at the Wuhan Union Hospital from January 2014 to December 2014 and evaluated its real-word clinical application using the hospital centralized monitoring method.A total of 848 cases were enrolled in this study.In these cases,it was mainly used for postoperative anti-inflammation and treating upper respiratory infection,pneumonia and bronchitis.Among them,39.86％ were contraindicated.Irregular medication of adults and children accounted for 1.91％ and 23.38％,respectively.Improper choice of solvent accounted for 3.18％.The choice of intravenous drip versus aerosol inhalation was reasonable.A case of adverse events (AEs) was observed in the monitoring period,and the incidence of adverse drug reaction (ADR) of ATES injection was 0.12％.ATES injection in our hospital is relatively safe with a low incidence of adverse reactions.The study assesses the clinical usage and adverse reactions of ATES injection,and provides suggestions for rational use in clinical practice.

Objective To investigate the effect of azelastin nasal spray combined with desloratadine in the treatment of allergic rhinitis .Methods Two hundred patients with allergic rhinitis were selected .According to the digital meter method ,the patients were randomly divided into observation group and control group ,with 100 cases in each group .The control group was treated by azelastin nasal spray , the observation group was given azelastin nasal spray combined with desloratadine .The clinical effects of the two groups were compared .Results The effective rate of the observation group(96.00%) was higher than that of the control group (80.00%),the difference was statistically significant(χ2 =6.235,P<0.05).After treatment,the scores of runny nose,nasal itching,nasal congestion,sneezing and the inferior turbinate swelling in the observation group were (1.1 ±0.2) points,(1.2 ±0.7) points,(1.1 ± 0.3)points,(0.8 ±0.3) points,(0.9 ±0.2) points,respectively,which were significantly lower than those in the control group [(1.4 ±0.9)points,(1.9 ±0.6)points,(1.8 ±0.8)points,(1.7 ±0.7)points,(1.9 ±0.9)points] (t=5.154,5.226,5.154,5.226,5.011,all P<0.05).Conclusion Azelastin nasal spray combined with deslorata-dine tablets in the treatment of allergic rhinitis can quickly relieve the patients 'clinical symptoms,improve the effec-tive rate,and it is safe and worthy of clinical popularization and application .

Objective To establish a Real-time Taqman probe technique system to detect the mtDNA 1555A > G mutation in deaf population. Methods Primers and Taqman probes for mtDNA 1555A > G mutation were designed and synthesized. The technique system for detecting mtDNA 1555A > G mutation using Real-time Taqman probes was established. Then the reliability of the technique was tested in 132 patients with severe to profound hearing loss who were detected for the mtDNA 1555A > G mutation by sequencing, Kit method and Real-time Taqman probe technique at the same time. Finally, the results by the above three ways were compared. Results Thirty-two cases with mtDNA 1555A > G mutation were found by the technique of Real-time Taqman probe. These findings coincided with the results from sequencing and Kit method completely. Both the false positive rate and the false negative rate were zero. Conclusions The technique possesses the merits of accuracy, conveniency, high sensitivity, high specificity and intuitionistic results, etc. Importantly, the Real-time Taqman probe technique only needs 1.5 hours to detect the 1555A > G mutation and it saves 4. 5 hours for one reaction compared with the Kit method popularly used nowadays. The technique system of detecting mtDNA 1555A > G mutation is reliable. It's suitable for large-scale detecting and preventive diagnosis of mtDNA 1555A > G mutation.

Objective To study the effect of carotid artery stenting (CAS) on rCBF and rCVR.Methods Seventeen patients with unilateral internal carotid artery symptomatic severe stenosis who underwent CAS in our hospital were included in this study.Their rCBF volume and rCVR were measured by single photon emission CT scanning combined with CO2 loading test 1 week be fore and 3 months after CAS.Their data were analyzed according to the ROI in ipsilateral middle cerebral artery blood supply territory.Results Sixty eight ROIs were detected in the 17 patients with impaired rCBF in 16 ROIs (23.5％) before CAS.The mean improved rate of rCBF was significantly higher in impaired rCBF and rCVR ROI before CAS than that of rCBF in normal and impaired rCVR ROI after CAS (P=0.001).The mean improved rate of rCVR was significantly higher in normal rCBF and impaired rCVR ROI after CAS than before CAS (P=0.014).The improved rate of rCBF was significantly higher in impaired rCBF and rCVR ROI after CAS than that of normal and impaired rCVR ROI before CAS (81.3％ vs 50.0％,P=0.027).The improved rate of rCVR was significantly higher in normal rCBF ROI and impaired rCVR ROI before CAS than in impaired rCBF and rCVR ROI after CAS (59.6％ vs 31.3％,P=0.047).Conclusion CAS can improve the ROI rCBF and rCVR in patients with unilateral ICA symptomatic severe stenosis.Its modified model is closely related with rCBF before CAS.

Objective To assess the diagnostic value of combined testing of urinary bladder cancer antigen(UBC),hyaluronic acid(HA)and survivin in the detection of bladder cancer.Methods This study included 64 bladder cancer patients and 20 urinary benign disease patients.The examinations of urine UBC by enzyme-linked immunosorbent assay(ELISA),HA by radioimmunology assay,survivin by RT-PCR and urine cytology were performed in them.Results The sensitivity of UBC(85.9%,55/64),HA (89.1%,57/64)and survivin(93.8%,60/64)was significantly higher than that of urine cytology (40.6%,P＜0.01).The specificity of UBC,HA,survivin and urine cytology was 85.0%(17/20),80.0% (16/20),95.0%(19/20)and 95.0%(19/20),respectively;there was no significant difference among these 4 methods(P＞0.05).The sensitivity of UBC,HA and survivin was also significantly higher than that of urine cytology in different histologic stages and grades(P＜0.05).The sensitivity of UBC and survivin was not significantly different among different histologie stages and grades(P＞0.05).With regard to HA test, the sensitivity in G_2 and G_3 groups was significantly higher than G_1 group(P＜0.01),but there was no differ- ence between G_2 and G_3 groups(P＞0.05);and no difference among different histologic stages(P＞0.05). However,the sensitivity of cytology was improved with the higher grade of bladder cancer(P＜0.01);there was no difference among histologic stages(P＞0.05),By combined use of UBC,HA and survivin,both the sensitivity and specificity were 100%.Conclusions The study indicates that UBC,HA and survivin are better diagnostic markers for the early detection of urinary bladder cancer.These tests are simple,feasible and noninvasive with higher sensitivity and specificity.In addition,combined use of them can improve the diag- nostic sensitivity and specificity.

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

Most patients with differentiated thyroid cancer have a good prognosis after radioiodine-131 therapy,but a small number of patients are insensitive to radioiodine-131 therapy and even continue to develop disease.At present,some targeted drugs can improve progression-free survival in patients with radioactive iodine-refractory differentiated thyroid cancer(RAIR-DTC),such as sorafenib and levatinib,have been approved for the treatment of RAIR-DTC.However,due to the presence of primary and acquired drug resistance,drug efficacy in these patients is unsatisfactory.This review introduces the acquired drug resistance mechanism of sorafenib in the regulation of mitogen-activated protein kinase(MAPK)and phosphatidylinositol-3-kinase(PI3K)pathways and proposes related treatment strategies,in order to provide a reference for similar drug resistance mechanism of sorafenib and effective treatment of RAIR-DTC.

OBJECTIVETo study the characteristics of spectra on proton magnetic resonance spectroscopy (1H-MRS) and its value in patients with mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS).

METHODSSeven clinically diagnosed patients with MELAS underwent magnetic resonance imaging (MRI) and 1H-MRS examinations. The 1H-MRS techniques, characteristics of the spectra, and its correlation with the laboratory tests were analyzed.

RESULTSCerebral abnormalities were revealed in all 7 patients on conventional MR images, and most abnormal signals were observed in bilateral occipital, parietal, and temporal lobes. We found 4 cases with basal ganglia involvement, 2 cases with mild frontal lobe lesions, and 1 case with involvement of lateral cerebral peduncles and thalami. Additionally, 1 patient was involved with left insular lobe. Spectra from prominent lesions in brain parenchyma showed lactate doublet peak in 6 patients, 3 of whom were also noted lactate peak in ventricular cerebrospinal fluid (CSF).

CONCLUSION1H-MRS may provide more direct information about the metabolism changes, which aids to affirm the diagnosis, and may replace the conventional invasive method of quantifying lactate in CSF.

Adolescent ; Adult ; Basal Ganglia ; pathology ; physiopathology ; Cerebral Cortex ; pathology ; physiopathology ; Child ; Female ; Humans ; Lactic Acid ; metabolism ; MELAS Syndrome ; diagnosis ; physiopathology ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Male ; Parietal Lobe ; pathology ; physiopathology

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.
Animals ; Caspase 8 ; biosynthesis ; genetics ; Cell Line ; Cyclin D1 ; biosynthesis ; genetics ; G1 Phase ; physiology ; Histones ; genetics ; metabolism ; Ki-67 Antigen ; biosynthesis ; genetics ; Male ; Mice ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Resting Phase, Cell Cycle ; physiology ; Spermatogonia ; cytology ; metabolism ; bcl-2-Associated X Protein ; biosynthesis ; genetics