The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

Objective:To investigate the value of multimodal MRI radiomics in the preoperative prediction of Fuhrman nuclear grade of clear cell renal cell carcinoma (ccRCC).Methods:A total of 129 patients with ccRCC confirmed by pathology from April 2011 to April 2021 in Third Affiliated Hospital of Soochow University were collected, and the imaging and clinicopathological data were retrospectively analyzed. All patients were divided into training set ( n=90) and validation set ( n=39) at the ratio of 7∶3 using random indicator method. According to the postoperative pathological results, Fuhrman grades Ⅰ and Ⅱ were included in the low grade group (96 cases, 65 cases in the training set and 31 cases in the validation set), and Fuhrman grades Ⅲ and Ⅳ were included in the high grade group (33 cases, 25 cases in the training set and 8 cases in the validation set). Two radiologists manually delineated regions of interest (ROI) on T 1WI, T 2WI, Dixon-water, Dixon-fat, susceptibility weighted imaging (SWI), blood oxygen level dependent (BOLD) images, and 396 texture features were extracted from each ROI. In the training set, intra-class correlation coefficient, Mann-Whitney U test, minimum redundancy maximum relevance and least absolute shrinkage and selection operator method were used to reduce the dimension of features to obtain the best texture features. The logistic regression was used to develop the multimodal radiomics model, and the receiver operating characteristic (ROC) curve was used to evaluate the effectiveness of the model in identifying high and low-grade ccRCC in training set and validation set. Results:Four SWI, one T 2WI and one BOLD texture features were selected for modeling. The areas under the ROC curve (95%CI) of the multimodal radiomics model for identifying high and low grade ccRCC in the training and validation sets were 0.859 (0.770-0.923) and 0.883 (0.740-0.964), with the specificity at 95.4% and 87.1%, the sensitivity at 68.0% and 87.5%, the accuracy at 87.8% and 87.2%, respectively. Conclusion:The multimodal MRI radiomics model based on T 2WI, SWI and BOLD images has high effectiveness in preoperative predicting Fuhrman nuclear grade of ccRCC.

OBJECTIVE: To assess the effect and safety of bloodletting puncture at hand twelve Jing-Well points (HTWPs) in acute stroke patients with conscious disturbance. METHODS: In this multi-center and randomized controlled trial, 360 patients suffered from ischemic or hemorrhagic stroke with conscious disturbance within 48 h from the onset of symptom were divided into bloodletting (180 cases) and control (180 cases) groups using a block randomization. Patients in both groups received routine Western medicine, and patients in the bloodletting group received additional bloodletting puncture at HTWPs on admission immediately before conventional treatment. The primary outcome measure was Glasgow Coma Scale (GCS) score and the secondary outcomes included blood pressure, respiratory rate and pulse rate. All variables were evaluated at baseline (before bloodletting), 0 (after bloodletting immediately), 15, 30, 50 and 80 min post bloodletting. RESULTS: At 80 min post bloodletting, the proportion of patients with improved consciousness in the bloodletting group was greater than the control group (P<0.05). In the separate analysis of moderate consciousness disturbance subgroup, bloodletting therapy benefited ischemic patients, and improved the eye and language response of GCS score at 15, 30, 50, 80 min post bloodletting (P<0.05 or P<0.01). No significant differences were observed regarding the secondary outcomes between two groups (P>0.05). CONCLUSION The bloodletting puncture at HTWPs was safe and could improve conscious levels of ischemic stroke patients, highlighting a first-aid intervention for acute stroke. (Registration No. ChiCTR-INR-16009530).

Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.

Objective To compare aortic root anatomical characteristics between severe aortic valve stenosis (AS) and aortic regurgitation (AR) patients, and to provide useful information for transcatheter aortic valve replacement (TAVR) device designs and procedural techniques for treatment of AR. Methods Consecutive patients admitted between April 2014 to May 2016 with severe AS or AR and planned to undergo transcatheter aortic valve replacement were included. There were a total of 57 AR and 113 AS patients. All patients underwent multi-detector computed tomographic imaging and echocardiography examinations. Results The mean aortic annulus diameter in AR patients was slightly but significantly larger than AS patients[ (26.4±3.7) mm vs. (25.2±2.9) mm, P=0.001]. The mean diameters of the ascending aorta[ (38.3±6.9) mm vs. (33.9±6.7) mm, P<0.001]and Valsalva sinus[ (38.9±6.9) mm vs. (32.7±4.5) mm, P<0.001] in AR patients were larger than in AS patients. The left coronary ostia height was of no significant difference between the 2 groups [ (12.5±3.7) mm vs. (13.4±3.2) mm, P=0.08] and the right coronary ostia height was higher in the AR group than in the AS group [ (17.5±5.0) mm vs. (15.3±3.3) mm, P=0.001]. Conclusions The anatomical aortic root data from patients with AS or AR in the present study may provide useful information for transcatheter aortic valve replacement device designs and procedural techniques for treatment of AR.

OBJECTIVE: To evaluate the effect of different triangular flap design and healing procedure on the sequelae after extraction of impacted lower third molars. METHODS: In this prospective, split-mouth study, 60 healthy patients with bilateral, symmetrically fully impacted lower third molars (LM3) were included, of whom 30 patients with totally bony impacted LM3 were allocated into group A, while the other patients with partially bony impacted LM3 were allocated into group B. All the teeth were extracted by the same surgeon. Triangular flap was used on one side, and the wound was primarily closed with two sutures (TF-P). On the other side, modified triangular flap was used with a triangular region of mucosa posterior to LM2 removed during operation, and a triangular soft tissue defect was left for drainage after suture (MTF-S). The patients were followed up on postoperative days 1, 3 and 7. Clinical parameters included postoperative pain, swelling, and trismus. Distal probing depth of adjacent second molar was assessed 6 months after extraction. Doctors responsible for the evaluation did not know the group and flap design. Paired sample t test was used to analyze the differences of postoperative sequelae between the two strategies. RESULTS: In group A, MTF-S strategy could reduce postoperative pain, ibuprofen consumption, and swelling significantly compared with TF-P strategy on the postoperative 1st and 3rd days (P<0.05). Besides, the trismus in the patients with TF-P strategy was more serious than that with MTF-S strategy on the postoperative 1st, 3rd and 7th days (P<0.05). However, statistic difference between the two strategies in pain, swelling and trismus was not detected in group B. Additionally, the VAS score in the patients with MTF-S strategy in group B increased slightly on the postoperative 4th day. The probing depth of the adjacent second molar was evaluated 6 months after extraction. Statistic difference was not detected between MTF-S strategy and TF-P strategy in the both groups. CONCLUSION Modified triangular flap with secondary healing procedure could effectively reduce the postoperative complications of totally bony impacted LM3. However, the difference between the two strategies in the probing depth of the adjacent second molar was not statically significant 6 months after extraction.
Edema ; Humans ; Mandible ; Molar, Third ; Pain, Postoperative ; Postoperative Complications ; Prospective Studies ; Surgical Flaps ; Tooth Extraction ; Tooth, Impacted

OBJECTIVETo explore the intrinsic connection between activation of classical nuclear factor-κB (NF-κB) pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells.

METHODSHuman lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines.

RESULTSGefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines (P<0.05 or 0.01). Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells (P<0.05) and HCC827 cells (P<0.01). The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines (P<0.05). In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions (P<0.05 or 0.01), and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65 (P<0.01 or 0.01).

CONCLUSIONThe activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.

Castleman Disease ; China ; Humans ; Syndrome

OBJECTIVETo analyze the clinical characteristics, image findings, laboratory examination, the therapeutic methods and clinical outcomes of bronchiolitis obliterans (BO) in pediatric patients.

METHODTwenty-six pediatric patients with BO were reported. All data were collected from cases who were hospitalized in the Department of Pediatrics, First Affiliated Hospital of Guangzhou Medical College from June 1(st), 2009 to the April 30(th), 2011, and infectious agents, clinical manifestations, risk factors, changes in imageology, laboratory examination, therapeutic methods and treatment responses were analyzed.

RESULTThe ranges of age at onset was 4.5 months-8 years in 26 cases (18 boys and 8 girls). The course of disease was (6.2 ± 3.5) months. The period of followed-up ranged from 2 to 24 months. The common clinical characteristics were persistent wheezing of different severity (26 cases, 100%), cough (24 cases, 92%), intolerance to exercise (22 cases, 85%), short of breath (21 cases, 81%), retraction (20 cases, 77%), wheezy phlegm (16 cases, 62%), keeping with crackles (10 cases, 38%), cyanosis around the mouth (3 cases, 12%) and no clubbed fingers (toes). In 18 cases the etiology was detected, mycoplasma (11 cases, 42%), respiratory syncytial virus (4 cases, 15%), parainfluenza virus (2 cases, 8%), influenza virus A (2 cases, 8%) and influenza virus B (2 cases, 8%), human bocavirus (HBoV) (1 case, 4%). There were 8 cases (31%) with combined infection. Chest X-ray in 10 cases indicated changes suggestive of bronchopneumonia (38%), in only 1 case there was an image of interstitial pneumonia disease (4%). All the patients were diagnosed by high-resolution computerized tomography (HRCT). All cases were demonstrated to have air retention, poor blood perfusion in lung, just like "Westemark sign" with HRCT. In 19 cases antineutrophil cytoplasmic antibody (ANCA) was determined and 10 patients (53%) were positive for P-ANCA, and 8 cases (42%) were positive for C-ANCA. All patients received oral corticosteroid and low doses azithromycin. In 13 cases (50%) the treatment effectively reduced the severity of disease and the frequency of cough and wheezing. The average number of days for symptom improvement was (7.1 ± 4.8) days.

CONCLUSIONRespiratory infection plays an important role in BO in children. The chronic and persistent wheezing, cough, intolerance to exercises, short breath, retraction were the main clinical manifestations. But these symptoms are non-specific. Chest X-ray can not provide enough information for diagnosis. Classical "Westemark sign" with HRCT is an important sign. ANCA with a high positive rate (approximately 50%) suppose immuno-lesion in BO. Oral corticosteroid and methotrexate may relieve clinical symptoms.

Bronchiolitis Obliterans ; diagnosis ; etiology ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Prognosis ; Treatment Outcome

ObjectiveAnalyze preoperative clinical relevanted factors of acute type A aortic dissection with hypoxemia according to a group clinical data.MethodsFrom January 2011 to June 2011,we have collected 54 preoperative cases of acute type A aortic dissection,including 42 males,12 females,aged 28-73 years old,onset to treatment time is 0.4-14.0 days.General information:age,gender,time of onset,body mass index,hypertension,diabetes mellitus,smoking,drinking,heart ejection fraction,prothrombin time,quantitative fibrinogen,fibrinogen degradation products,D-dimer,C-reactive protein,procalcitonin,ICU time,length of hospital stay.According to the blood gas analysis of quiet state case without oxygen,with PaO2 ＜ ( 100-age ×0.33 ±5) mm Hg is for the hypoxemia group,equal or higher than this is no-hypoxemia group.ResultsNo-hypoxemia group has 14 cases,11 males,3 females,average aged (51.14 ± 14.24) years old,including 12 operation patients ( no death) and 2 no-operation patients(2 cases death).Hypoxemia group has 40 cases,31 males,9 females,average aged (50.53 ± 9.73 ) years old,including 33 operation patients(2 cases death) and 7 no-operation patients(7 cases death).There is no significant difference in age,gender,time of onset,hypertension,diabetes mellitus,smoking,drinking,cardiac ejection fraction,prothrombin time and fibrinogen.There is statistically significant on body mass index,fibrinogen degradation products,D-dimer,C-reactive protein,procalcitonin,ICU time and length of hospital stay time ( P ＜ 0.05 ).ConclusionPreoperative hypoxemia with acute type A aortic dissection is associated with obesity,excessive inflammation and activation of coagulation and fibrinclytic system,and hypoxemia may prolong the time of operative patients with acute type A aortic dissection in ICU and hospital.