Bacillus mycoides JA20-1 was screened and identified as a biocontrol bacterium with a high capacity for producing volatile organic compounds(VOCs) in the laboratory. This strain had significant inhibitory effects on various postharvest disease pathogens in crops, such as Botrytis cinerea, as well as soil-borne disease pathogens in ginseng, such as Sclerotinia ginseng. In order to accelerate its industrialization process, in this study, single-factor experiments and response surface optimization methods were used. The fermentation medium and fermentation conditions in the shake flask of strain JA20-1 were systematically optimized by using cell production volume as the response variable. Meanwhile, the biocontrol effect of JA20-1 on B. cinerea of ginseng during the storage period was evaluated by using the method of fumigation in a dry dish in vitro. The results indicated that the optimal fermentation medium formulation for strain JA20-1 was as follows: 1% yeast paste, 1% soluble starch, 0.25% K_2HPO_4·3H_2O, and 0.2% NaCl. The optimal fermentation conditions in the shake flask were vaccination size of 3%, culture volume of 50 mL in a 250 mL Erlenmeyer flask, pH of 6.2, fermentation temperature of 34 ℃, shaking speed of 180 r·min~(-1), and incubation time of 18 hours. The bacteria count in the fermentation broth under these conditions reached 2.17 × 10~8 CFU·mL~(-1), which was 6.58 times higher than before. The average control efficacy of the fermentation broth on Botrytis cinerea of ginseng under in vitro fumigation reached 61.70% and 84.04% respectively, when 20 mL and 30 mL per dish were used. The research provided theoretical support and technical foundation for the development and utilization of Bacillus mycoides JA20-1 and the biocontrol of soil-borne diseases in ginseng and postharvest diseases in crops.
Botrytis/drug effects* ; Fermentation ; Panax/microbiology* ; Plant Diseases/prevention & control* ; Volatile Organic Compounds/metabolism* ; Bacillus/physiology* ; Pest Control, Biological/methods* ; Biological Control Agents/metabolism* ; Culture Media/chemistry*

ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang （BWT） on non-alcoholic fatty liver disease （NAFLD） and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups： normal control， model， positive drug （pioglitazone hydrochloride 1.95×10^-3 g·kg^-1）， and low-， medium-， and high-dose BWT （1.3，2.5 and 5.1 g·kg^-1）. Following a 12-week high-fat diet （HFD） inducement， the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week， glucose tolerance （GTT） and insulin tolerance （ITT） tests were conducted. Subsequently， the mice were euthanized for the collection of liver tissue and serum， and the subcutaneous adipose tissue （iWAT） and epididymal adipose tissue （eWAT） were weighed. Serum levels of total triglycerides （TG） and liver function indicators，such as alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， were determined. Histological examinations， including oil red O staining， hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy， were performed to evaluate hepatic lipid deposition， pathological morphology， and ultrastructural changes， respectively. Meanwhile， Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were employed to analyze alterations， at both gene and protein levels， the insulin signaling pathway molecules， including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 （IRS1/2/Akt/FoxO1）， glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase （Pepck） and glucose-6-phosphatase （G6Pase）， lipid metabolism-related genes stearoyl-coA desaturase-1 （SCD-1） and carnitine palmitoyltransferase-1 （CPT-1）， fibrosis-associated molecules α-smooth muscle actin （α-SMA）， type Ⅰ collagen （CollagenⅠ）， and the fibrosis canonical signaling pathway transforming growth factor-β₁/drosophila mothers against decapentaplegic protein2/3（TGF-β₁/p-Smad/Smad2/3）， inflammatory factors such as interleukin（IL）-6， IL-8， IL-11， and IL-1β， autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1， and the expression of mammalian target of rapamycin （mTOR）. ResultCompared with the model group， BWT reduced the body weight and liver weight of NAFLD mice（P<0.05， P<0.01）， inhibited liver lipid accumulation， and reduced the weight of white fat： it reduced the weight of eWAT and iWAT（P<0.05， P<0.01） as well as the serum TG content（P<0.05， P<0.01）. BWT improved the liver function as reflected by the reduced ALT and AST content（P<0.05， P<0.01）. It improved liver insulin resistance by upregulating IRS2， p-Akt/Akt， p-FoxO1/FoxO1 expressions（P<0.05）. Besides， it improved glucose and lipid metabolism disorders： it reduced fasting blood glucose and postprandial blood glucose（P<0.05， P<0.01）， improved GTT and ITT（P<0.05， P<0.01）， reduced the expression of Pepck， G6Pase， and SCD-1（P<0.01）， and increased the expression of CPT-1（P<0.01）. The expressions of α-SMA， Collagen1， and TGF-β₁ proteins were down-regulated（P<0.05， P<0.01）， while the expression of p-Smad/Smad2/3 was downregulated（P<0.05）， suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6， IL-8， IL-11 and IL-1β（P<0.01） and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression（P<0.05）while downregulating the expression of p62/SQSTM1 and mTOR（P<0.05）. ConclusionBWT ameliorates NAFLD by multifaceted improvements， including improving IR and glucose and lipid metabolism， anti-inflammation， anti-fibrosis， and enhancing autophagy. In particular， BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.

Objective To establish a nomogram model for efficiently predicting overall survival(OS)and cancer-specific survival(CSS)in patients with CRCHBM.Method 2239 patients from 2010 to 2019 were retrospectively analyzed from the Surveillance,Epidemiology,and End Results Program(SEER)databases and Wuhan Union Hospital Cancer Center.SEER is randomly assigned to the training and internal validation cohorts,and the Wuhan database serves as the external validation.Cox regression analyses were used to determine the independent clinicopathological prognosis factors affecting OS and CSS,and a nomogram was constructed to predict OS and CSS.The clinical utility of columnar plots was assessed using calibration curves,area under the curve(AUC),and decision curve analysis(DCA).Result OS column line graphs were constructed based on nine independent predictors:age,tumor location,degree of differentiation,tumor size,TNM stage,chemotherapy,primary focus surgery,number of lymph nodes sampled,and serum carcinoembryonic antigen(CEA)level.The C-index of the nomogram to predict the 1-,3-,and 5-year OS were 0.764,0.790,and 0.805 in the training group,0.754,0.760,and 0.801 in the internal validation group,and 0.822,0.874,and 0.906 in the external validation group.CSS column line graphs were constructed based on 3 independent predictors of TNM staging,radiotherapy and chemotherapy.The 1-,3-,and 5-year CSS AUROC values of the training group were 0.791,0.757,and 0.782,respectively.0.682,0.709,0.625 in the internal validation group and 0.759,0.702,0.755 in the external validation group,respectively.The results of receiver operating characteristic curve(ROC),ROC and DCA showed that the use of our model was more effective in predicting OS and CSS than other single clinicopathological features.Conclusion In summary,the nomogram based on significant clinicopathological features can be conveniently used to predict OS and CSS individually in patients with CRCHBM.

Objective:To investigate the clinical effects and indications of tibial condylar valgus osteotomy (TCVO) in treating varus unicompartmental knee osteoarthritis.Methods:A retrospective analysis was conducted in 32 patients (45 knees) who suffered from varus unicompartmental knee osteoarthritis and underwent TCVO from June 2016 to June 2018. These patients were aged 65.8±8.3 (range from 52 to 79) years, including 12 males (18 knees) and 20 females (27 knees). All enrolled individuals presented obvious expansion of the lateral joint space with joint line convergence angle (JLCA) of 7.19°±2.69°. Based on the full-length standing X-ray imaging of the lower limbs at before and 2 years after surgery, the percentage of mechanical axis (%MA), femorotibial angle (FTA), hip-knee-ankle angle (HKA), lateral distal femoral angle (LDFA) and medial proximal tibial angle (MPTA) were measured and analyzed to evaluate the improvements of lower extremity alignments. The medial tibial plateau depression (MTPD), posterior proximal tibial angle (PPTA), JLCA and joint space width (JSW) were measured and analyzed to evaluate the congruency of the knee joint and shape of the tibial plateau based on positive and lateral radiographs of knee joint. In addition, visual analogue scale (VAS) and Western Ontario and McMaster Universities (WOMAC) score were evaluated to assess the clinical effects of TCVO pre-operatively and at 1 year or 2 years after surgery.Results:All patients were followed up for 33.4±7.4 (range from 25 to 40) months. Comparing to the preoperative radiological data, %MA at 2 years after surgery increased from 3.78%± 14.34% to 66.16%±9.90%, FTA from 185.41°±4.45° to 170.81°±2.87°, HKA from 169.69°±1.70° to 181.16°±2.39°, MPTA from 83.03°±3.20° to 90.84°±3.67° all with statistical significance ( P<0.05). There was no significant difference for PPTA between before (89.22°±1.52°) and 2 years (88.97°±1.57°) after surgery ( t=0.638, P=0.526). MTPD improved from -7.81°±3.27° to 5.78°±2.19° ( t=19.218, P<0.001). However, there was no significant difference for PPTA between before (81.63°±3.28°) and 2 years (82.25°± 2.21°) after surgery ( t=0.881, P=0.382). JLCA reduced from 7.19°±2.69° to 0.22°±2.09°. The medial and lateral JSW were corrected from 2.45±0.23 mm and 5.86±0.25 mm to 3.73±0.27 mm and 4.68±0.34 mm ( P<0.05), respectively. Additionally, VAS and WOMAC scores improved from 6.46±2.21 and 52.66±16.69 preoperatively to 2.94±1.72 and 19.31±14.87 at 1 year after surgery, and to 1.39±1.45 and 13.66±15.44 at 2 years after surgery, respectively ( P<0.05). Conclusion:Satisfactory early therapeutic outcomes could be achieved by TCVO in varus unicompartmental knee osteoarthritis with subluxated lateral joint and increased JLCA. TCVO can correct intra-articular varus deformity, adjust mechanical axis and relieve knee joint pain and dysfunction.

Objective To explore the operative technique and clinical results of posterior tibial artery perforator flap within saphenous nerve branch for sensory reconstruction.Methods From January,2016 to June,2018,9 patients suffered from soft tissue defect were treated by the posterior tibial artery perforator flap containing saphenous nerve branch.Seven patients were males and 2 were females,with age ranged from 31 to 62 years.Soft tissue defects located in hands in 5 patients,plantar in 2 patients,ankle in 1 patient and dorsal foot in 1 patient.The size of soft tissue defects ranged from 8.0 cm×2.5 cm to 21.0 cm×4.0 cm.The regular post-operative followed-up was performed.Results All flaps survived without complications.The size of flap ranged from 10.0 cm×3.5 cm-23.0 cm×5.0 cm.Donor sites were primarily closed in 5 patients and secondary closed in 4 patients.Followed-up ranged from 6 to 15 months with 10 months in average.The contour of flaps were satisfied and the sensory function of the donor sites were normal.At 6 months followed-up,SW test reached 5.07 in all flaps,and 2PD ranged from 14 to 35 mm.Conclusion The novel sensory flap can provide satisfied sensory outcome without sacrificing main artery and saphenous nerve,and is a good candidate for sensory reconstruction of soft tissue defects.