L-valine is an important essential branched-chain amino acid widely used in industries such as feed, pharmaceuticals, and food. In order to further enhance the production performance of L-valine, this study systematically engineered the metabolism of a Corynebacterium glutamicum strain, preserved in the laboratory, which is capable of producing L-valine. First, strain VH-9 was obtained by enhancing the precursor supply, synthesis pathway, and transport system of L-valine. In a 5 L fermenter, the titer, yield, and productivity of L-valine were 76.6 g/L, 0.45 g/g, and 2.39 g/(L·h), respectively. Furthermore, strain VH-18 was obtained by enhancing the uptake of substrate glucose and balancing energy supply to reduce succinate accumulation, with the titer, yield, and productivity of L-valine increased to 82.7 g/L, 0.52 g/g, and 2.58 g/(L·h), respectively. After optimization of fermentation conditions, the titer, yield, and productivity of L-valine in strain VH-18 were further improved to 88.7 g/L, 0.54 g/g, and 2.77 g/(L·h), respectively. This study has achieved the high-efficiency production of L-valine through a systems metabolic engineering strategy.
Corynebacterium glutamicum/genetics* ; Metabolic Engineering/methods* ; Valine/biosynthesis* ; Fermentation ; Glucose/metabolism*

A 26-year-old female patient was admitted to the hospital with recurrent infection of the incision after resection of a back sebaceous cyst,and the pus culture showed Mycobacterium abscessus.Clinical pharmacists reviewed relevant guidelines and literature,analyzed and summarized drug selection,drug resistance,adverse drug reactions and coping strategies,sequential treatment plans and treatment courses,and assist physicians in formulating individualised anti-infective treatment plans.Initially imipenem,amikacin and azithromycin were given according to bacterial culture results.Secondly,according to the results of drug sensitivity,they were changed to tigecycline,amikacin and clarithromycin.Finally,due to the adverse drug reaction of tigecycline and the recurrence of sinus in the patient,and considering the possibility of imipenem and clarithromycin resistance,the anti-infection regimen was adjusted in time to cefoxitin,amikacin and azithromycin.During treatment,the clinical pharmacist monitors drug effectiveness and adverse reactions,combining pharmacy expertise with clinical practice.After the treatment,the patient improved and was discharged.After discharge,azithromycin and omacycline were successively given to continue anti-infection treatment,and the incision on the patient's back basically healed through follow-up.Clinical pharmacist involvement in Mycobacterium abscessus anti-infective drug selection can provide individualised drug regimens for patients,ensure the safety and effectiveness of medication for patients,and provide references for the treatment and management of similar patients.

Objective: To investigate the clinical characteristics during the perioperative period of pheochromocytoma in patients aged 60 years and over. Methods: Data of age, sex, tumor size, anesthesia time, intraoperative bleeding volume, intraoperative blood pressure, complications and hospitalization time from patients with pheochromocytoma in our hospital treated by the retroperitoneal laparoscopic adrenalectomy from January 2008 to October 2018 were retrospectively analyzed.The relationships of age with the intraoperative hemodynamic instability and postoperative complications were analyzed. Results: A total of 203 patients with pheochromocytoma met the inclusion criteria were enrolled.Age over 60 years(OR1.771, 95%CI=1.015-3.089, P=0.044)was an independent risk factor for intraoperative hemodynamic instability. Conclusions Laparoscopic retroperitoneal adrenalectomy is a safe surgical method for pheochromocytoma patients aged 60 years and over.For elderly patients with pheochromocytoma, especially those with a tumor diameter of more than 5 cm, special attention should be paid to the prevention of intraoperative hypertension crisis.

Objective To approach the spine MRI features and its possibility of etiology for refractory lower urinary tract symptoms(LUTS) in female patients.Methods We conducted prospectively a cross sectional description study of female patients with refractory LUTS during January 16 through March 27 in 2017 based on a urologist's outpatient work.The including criteria were adult female patients with refractory LUTS which defined as having LUTS more than three months and having poor response to behavior therapy and medication treatment.The excluding criteria were patients having evidence of infection,tumor,stone in urinary tract,any central nerve system diseases,or any other diseases may potentially producing LUTS.Data collected included patients demographic information,main complains,present disease features,disease history,physical examination,urine routine,urodynamic study and spine MRI.The characteristics of clinical manifestation,urodynamic study and spine MRI were analyzed.Results During the time span of study,totally 70 cases had been diagnosed as having refractory LUTS and had qualified data of clinical recordings,urodynamic study and spine MRI.Among these 70 cases,63 (90.9％) had storage phase symptoms,11 (15.7％) had voiding phase symptoms,8 (11.4％) had postmicturition symptoms,12 (17.1％) also had disorders in defecating,45 (64.3％) had pain in lower abdomen or pelvic region.69 cases (98.6％) had urodynamic disorders,33 (47.1％) had oversensitivity of bladder,12 (17.1％) had smaller bladder volume,16(22.9％) had detrusor overactivity,15 (21.4％) had bladder outlet obstruction,39(55.7％) had detrusor underactivity.69 cases(98.6％)had spine MRI abnormalities,54(77.1％) had sacral nerve lesions,49 (70.0％) had cervical lesions,48 (68.6％) had lumbar lesions,4 had thorathic lesions.Conclusions The present study revealed extraordinary high prevalence of abnormality in urodynamic parameters and spine MRI in female patients with refractory LUTS,which implies possibility that the refractory LUTS are caused by lesions in spinal nerve system.

Objective To study the effect of different atorvastatin doses on contrast-induced nephropathy (CIN) and the role of neutrophil gelatinase-associated lipocalin (NGAL) in early diagnosis of CIN after PCI.Methods Two hundred and twelve patients admitted to our hospital for PCI from June 2015 to January 2017 were divided into high atorvastatin dose treatment group as group A (n=108) and conventional atorvastatin dose treatment group as group B (n=104).They underwent coronary arteriography and PCI.Their serum creatinine and urinary NGAL levels were measured before and at 4,24,48,72 h respectively after coronary arteriography and PCI.The role of urinary NGAL level in early diagnosis of CIN was analyzed according to its ROC curve.Results CIN occurred in 6 patients of group A and in 19 patients of group B with an incidence of 5.6％ and 18.3％ respectively (P=0.012).The urinary NGAL level was significantly higher in two groups at 4,24,48,72 h respectively after coronary arteriography and PCI than before coronary arteriography and PCI,reached its peak at 72 h,and was significantly higher in group B than in group A (P＜0.05).The area under the ROC curve was 0.917,0.919,0.928,0.953 respectively at 4,24,48,72 h after coronary arteriography and PCI.The sensitivity and specificity of urinary NGAL in early diagnosis of CIN were 92.10％ and 100.00％ respectively.Conclusion High atorvastatin dose treatment can reduce the occurrence of CIN.Urinary NGAL level can indicate the change of renal function earlier than serum creatinine level after PCI,and can thus be used as an early predictor of CIN.

In a previous study, we screened thousands of long non-coding RNAs (lncRNAs) to assess their potential relationship with congenital heart disease (CHD). In this study, uc.4 attracted our attention because of its high level of evolutionary conservation and its antisense orientation to the CASZ1 gene, which is vital for heart development. We explored the function of uc.4 in cells and in zebrafish, and describe a potential mechanism of action. P19 cells were used to investigate the function of uc.4. We studied the effect of uc.4 overexpression on heart development in zebrafish. The overexpression of uc.4 influenced cell differentiation by inhibiting the TGF-beta signaling pathway and suppressed heart development in zebrafish, resulting in cardiac malformation. Taken together, our findings show that uc.4 is involved in heart development, thus providing a potential therapeutic target for CHD.

Objective To investigate the efficacy and safety of transcather arterial embolization (TAE) plus sirolimus for the treatment of refractory Kasabach-Merritt syndrome (KMS) in infants. Methods Clinical data of twelve infants with refractory KMS treated between December 2015 and October 2016 in a single hospital were retrospectively analyzed. TAE were performed in all patients after failed traditional multiple therapies, followed by oral sirolimus administration. The dose of sirolimus was modulated according to the level of sirolimus, the count of platelet, the shrinkage of the lesion and the side effects, which were monitored regularly during the study. Results All 12 patients were treated with TAE plus sirolimus therapies successfully. The platelet count for all patients increased to≥100×109/L for the first time at (7±5) days. Stabilization of platelet level was obtained in (15±7) days averagely. Before the treatment, two infants had a normal fibrinogen level and the fibrinogen level in the other 10 infants was found to be increased to≥2.0 g/L at (9 ± 4)days for the first time and was then stabilized at levels>2.0 g/L at (19 ± 7)days after treatment. One patient showed skin fester (GradeⅡ), one patient had a fever with acute pulmonary infection (Grade Ⅲ) and both patients improved well after symptomatic treatment. There were no serious complications in the other ten patients. Conclusions TAE plus sirolimus can rapidly improve levels of platelets and fibrinogen, and it is a safe, useful and effective method for treatment of refractory KMS in infants.

Objective To evaluate the therapeutic effects and safety of transcatheter arterial sclerosing embolization (TASE) combined with oral Propranolol in infantile huge hemangiomas.Methods A total of 76 infants with huge hemangioma were treated by TASE.All patients received oral Propranolol based on their weight.The follow-up and efficacy evaluation were performed from the last treatment.And the complications were analyzed.Results Seventy-six cases underwent 95 case-times of TASE totally.The success rate of puncture technique was 100％ (95/95).All patients were divided into 5 levels (0-Ⅳ levels) based on the curative effect of the follow-up,including 6 cases (6/76,7.90％) of level Ⅰ,19 cases (19/76,25.00％) of level Ⅱ,23 cases (23/76,30.26％) of level l,and 28 cases (28/76,36.84％) of level Ⅳ.The total effective rate was 100％ (76/76).There was no serious complications,and the curative effect was stable during the follow-up.Conclusion TASE combined with oral Propranolol in infantile huge hemangiomas is effective with less invasiveness and fewer complications.

Objective To investigate the efficacy and safety of transcather arterial embolization (TAE) plus sirolimus for the treatment of refractory Kasabach-Merritt syndrome (KMS) in infants. Methods Clinical data of twelve infants with refractory KMS treated between December 2015 and October 2016 in a single hospital were retrospectively analyzed. TAE were performed in all patients after failed traditional multiple therapies, followed by oral sirolimus administration. The dose of sirolimus was modulated according to the level of sirolimus, the count of platelet, the shrinkage of the lesion and the side effects, which were monitored regularly during the study. Results All 12 patients were treated with TAE plus sirolimus therapies successfully. The platelet count for all patients increased to≥100×109/L for the first time at (7±5) days. Stabilization of platelet level was obtained in (15±7) days averagely. Before the treatment, two infants had a normal fibrinogen level and the fibrinogen level in the other 10 infants was found to be increased to≥2.0 g/L at (9 ± 4)days for the first time and was then stabilized at levels>2.0 g/L at (19 ± 7)days after treatment. One patient showed skin fester (GradeⅡ), one patient had a fever with acute pulmonary infection (Grade Ⅲ) and both patients improved well after symptomatic treatment. There were no serious complications in the other ten patients. Conclusions TAE plus sirolimus can rapidly improve levels of platelets and fibrinogen, and it is a safe, useful and effective method for treatment of refractory KMS in infants.

In order to obtain nucleotides aptamers bind to IgE, 80 bp nucleotides single-stranded DNA library containing 40 random nucleotides was designed and synthesized. Oligonucleotides that bind to human Cepsilon3-Cepsilon4 protein were isolated from ssDNA pools by the systematic evolution of ligands by exponential enrichment (SELEX) method using nitrocellulose filters as screening medium. Through the optimization of critical PCR and asymmetric PCR parameters including annealing temperature, cycles, and molar ratios of target protein and ssDNA etc, a suitable screening system was established. The aptamers of Cepsilon3-Cepsilon4 protein with high affinity and high specificity were identified by ELISA with biotin-streptavidin-horseradish peroxidase system, and its primary sequence and second structure were analyzed by DNAMAN package and DNA folding sever after being cloned and sequenced. Moreover, target protein was bound to one aptamer and another aptamer modified with biotion together forming a sandwich-like complex, which was captured in microwell to detect IgE concentration using the optimal combination in the sandwich method named enzyme-linked aptamers sorption assay (ELASA). The method could be used for the quantitative detection of human IgE, and whose sensitivity reached to 120 ng x mL(-1).