Flowering and bolting are important agronomic traits in cruciferous crops such as Brassica juncea. Timely flowering can ensure the crop organ yield and quality, as well as seed propagation. The WRKY family plays an important role in regulating plant bolting and flowering, while the function and mechanism of WRKY12 in B. juncea remain unknown. To explore its function and mechanism in bolting and flowering of B. juncea, we cloned and characterized the BjuWRKY12 gene in B. juncea and found that its expression levels were significantly higher in flowers and inflorescences than in leaves. BjuWRKY12 belonged to the Ⅱc subfamily of the WRKY family, and subcellular localization indicated that the protein was located in the nucleus. Ectopic overexpression of BjuWRKY12 in transgenic lines promoted bolting and flowering, leading to significant increases in the expression levels of flowering integrators SOC1 and FUL. Furthermore, yeast one-hybrid and dual luciferase reporter system assays confirmed that BjuWRKY12 directly bound to the promoters of BjuSOC1 and BjuFUL, undergoing protein-DNA interactions. This discovery gives new insights into the regulation network and molecular mechanisms of BjuWRKY12, laying a theoretical foundation for the breeding of high-yield and high-quality varieties of B. juncea.
Mustard Plant/metabolism* ; Flowers/growth & development* ; Plant Proteins/physiology* ; Promoter Regions, Genetic/genetics* ; Gene Expression Regulation, Plant ; Plants, Genetically Modified/genetics* ; Transcription Factors/metabolism* ; MADS Domain Proteins/metabolism*

Objective:To discuss the long-term survival and risk factors of thyroid cancer in the real world in China.Methods:The clinical data of thyroid cancer patients who underwent initial surgery from Apr. 1998 to Dec. 2018 were retrospectively analyzed, including patients’sex, age, surgical records, pathology, hospitalization records and follow-up. According to the prognosis, the patients were divided into disease-free survival group and recurrence/metastasis/death group. Univariate analysis and multivariate regression analysis were conducted to analyze the risk factors affecting the prognosis of thyroid cancer. The clinical features and prognostic risk factors of thyroid cancer patients were investigated.Results:A total of 2038 cases were collected, and the longest follow-up time was more than 20 years. A total of 1876 cases were included in the study, 162 cases were lost, and the rate of follow-up was 7.9%. Among them, 1858 survived, the overall survival rate was 99.04%; 18 died, and the overall mortality rate was 0.96%. According to the prognosis of thyroid cancer, the patients were divided into 2 groups, including 1808 cases in the disease-free survival group and 68 cases in the relapsed-metastatic-death group. The study found that there were statistical differences between the two groups in terms of patients’age [ (45.40±11.016) vs (51.53±15.199, P=0.000) , the male ratio (32.854%, 48.529%, P=0.001) , whether tumor breaks through capsule (20.077%, 33.823%, P=0.006) , central lymph node metastasis (48.834%, 70.588%, P=0.001) and lateral lymph node metastasis (31.084%, 55.882%, P=0.000) , and there was no difference between the number of tumor lesions. Conclusion:Thyroid cancer has a good prognosis, but according to the characteristics of patients with thyroid cancer in my country, it should still be treated early in the clinic, and the standardization and thoroughness of surgery should be adhered to during the treatment.

Malunion is a common complication following a calcaneal fracture which was not treated or treated inappropriately.It is a therapeutic target and a great challenge as well to relieve pain, correct deformity and restore the function of the affected foot in clinical treatment of calcaneal malunion. As a result of researches by scholars at home and abroad focusing on the biomechanical mechanisms underlying the symptoms caused by calcaneal malunion, a variety of corrective calcaneal osteotomy has been widely applied in clinical practice to specifically correct the calcaneal deformity and restore normal calcaneal morphology. This review expounds on the techniques, outcomes, indications and complications of corrective calcaneal osteotomies commonly used in clinic.

Incidence of acute closed Achilles tendon rupture is high in clinic. Accurate diagnosis of this injury depends on a combination of physical signs and imaging examinations. Although conservative and surgical interventions are common for acute closed Achilles tendon rupture, its optimal treatment remains controversial. This literature review focuses on the research progress concerning this injury so as to analyze its risk factors, diagnostic methods, treatments, early functional rehabilitation, postoperative complications and prognosis evaluation.

Objective:To analyze the incidence of HIV infection and identify associated risk factors in drug abusers among men who have sex with men (MSM) in Tianjin and Shanghai through a cohort study and provide a basis for HIV prevention in this population.Methods:A prospective cohort study was conducted among MSM who had a history of drug abuse in the past six months from June 2016 to June 2018 in the two cities. MSM were investigated to obtain information on sociodemographic characteristics, HIV-related sexual behaviors, and drug abuse. Follow-up investigation and HIV testing were carried out based on the baseline survey and testing. Cox regression analysis was conducted to identify the risk factors for HIV infection.Results:There were 455 eligible subjects, and 16 new HIV infection cases were identified in the 2-year follow-up survey. The cumulative follow-up time was 586.08 person-years, and the incidence of HIV infection was 2.73/100 person-years. The multivariate Cox regression analysis results showed that compared with those aged ≥25 years, consistent condom use during anal sex with men in the past six months, without mixed-use of drugs, these aged <25 years ( HR=5.01, 95% CI: 1.09-23.11), inconsistent condom use during anal sex with men in the past six months ( HR=1.58, 95% CI: 1.04-2.41) and mixed-use of drugs ( HR=1.92, 95% CI: 1.08-3.40) were significantly associated with HIV infection in this cohort. Conclusions:The younger age, inconsistent condom use during anal sex with men, and mixed drug use appeared as risk factors of new HIV infection in drug abusers among MSM. HIV prevention and intervention in this population should be further strengthened.

OBJECTIVE：To provide reference for elucidating the anti-allergic asthma constituents in alkaloids-free part of Ephedrae Herba. METHODS ：Ephedrae Herba was extracted with 85％ ethanol and n-heptane，and then subjected to solid-phase extraction（filler AC 18）for pretreatment to enrich alkaloids-free part from the extract of Ephedrae Herba. HPLC method was adopted，and alkaloids-free fractions of Ephedrae Herba were performed on Unitary C 18 column and Eclipse XDB-C 18 column. Using high expression G protein coupled-receptor 35（GPR35 receptor）in HT- 29 cell as target ，GPR35 receptor agonist zaprinast （1 μmol/L）as positive control ，DMR response value as the detection index ，the agonistic and desensitizing activity of each fraction（100 μg/mL）on GPR 35 receptor was screened by label-free integrated pharmacological method ，so as to screen active anti-allergic asthma fraction. HPLC-Q-TOF-MS method was used to identify the chemical composition of the selected active fractions. RESULTS ：The alkaloids-free part of Ephedrae Herba was divided into two parts ，involving the precipitated part before solid phase extraction and the 95％ methanol elution part ；from them ，20 fractions were screened. Among them ，the precipitated fraction F 1.5-F1.10 and 95％ methanol eluted fraction F 2.5-F2.10 had a strong agonistic activity on GPR 35 receptor；at the same time，GPR35 receptor agonist zaprinast showed a relatively strong desensitization activity. The signal intensity of DMR induced by F1.5-F1.10 in the precipitated part of HT- 29 cells was even higher than that of reference drug zaprinast. By HPLC-Q-TOF-MS analysis，24 chemical components were identified from active fractions ，involving 14 flavonoids，2 volatile oils ，7 organic carboxylic acids ，1 anthraquinones. CONCLUSIONS ：The alkaloid-free part of Ephedrae Herba is mainly flavonoids and has anti-allergic asthma activity.

Objective: To investigate the surgical treatment strategies for persistent and recurrent parathyroidectomy after total parathyroidectomy plus autotransplantation(tPTX+ AT) in cases of renal hyperparathyroidism. Methods: From Oct 2009 to Oct 2018, 480 patients with renal hyperparathyroidism received tPTX+ AT in our hospital. 32 patients suffered from post-op persistent (31) and recurrent (1) hyperparathyroidism .The high frequency ultrasonography combined with fine needle puncture eluent PTH determination and SPECT/CT co-computed tomography fusion imaging were used as qualitative and localizing diagnostic methods before reoperation. Results: Of the 32 patients, 28 cases underwent reoperation for once, and 4 underwent reoperations fort wice. 36 parathyroid glands and 2 grafts were resected.In 31 patients the bone pain, skin pruritus relieved significantly or disappeared, and muscle strength gradually increased compared with that before operation. Level of iPTH in 31 patients fluctuated between 15 and 90 ng/L. Postoperative parathyroid hormone decreased compared with that before the operation (P<0.05). One patient was still with persistent hyperparathyroidism despite reoperation, whose iPTH fluctuated between 550 and 1 000 ng/L during 6 months to 2 years follow-up. Conclusion Ultrasonography, FNA-iPTH and SPECT/CT co-computed tomography can be used as methods for qualitative and localizing diagnosis of PTPT or PHPT.With accurate preoperative localization, resection of all parathyroid glands is an effective treatment for posto perative persistent and recurrent renal hyper parathyroidism.

OBJECTIVE： To screen potential eEF2K inhibitor molecules， and to provide reference for the design and R&D of eEF2K inhibitor. METHODS： The eEF2K crystal structure model was constructed by homology modeling technique. The model was optimized by Loop optimization and molecular dynamics. With the help of SAVES online server， the above models were evaluated from three aspects such as Verify_3D， EERAT and Laplace diagram. Totally 55 eEF2K inhibitor molecules were collected. Hypogen pharmacophore model with activity prediction ability was constructed based on 28 of them （odd number， as training set） by Insight Ⅱ software and validated by other 27 （even number， as test set）. The optimal pharmacophore model was screened by fitting the predicted and experimental values of activity [i.e. negative logarithm of half inhibitory concentration （pIC50）] and using Ligand profiler thermogram. The virtual screening of small molecules of eEF2K inhibitors was carried out by combining the above pharmacophore model， Lipinski’s five rules and molecular docking method. RESULTS & CONCLUSIONS： The overall quality factor score of the crystal structure model of eEF2K protein was 93.697. Among them， 83.33％ of the amino acid Verify_3D score was more than or equal to 0.2， and 1.7％ of the total amino acids were located in the non-permissible region. The amino acid conformation and skeleton structure of the model were reasonable and the reliability of the model was high. Totally 9 Hypogen pharmacophore models （No. 02-10） with active predictive function were constructed， among which No. 03 pharmacophore model included 2 hydrogen bond receptors and 2 conjugated aromatic rings， which could better distinguish active and inactive molecules. The predicted value of pIC50 fitted the experimental value best （the correlation coefficient was 0.665 3）， and it had good predictive ability and high reliability. Finally， 9 potential eEF2K inhibitor molecules were obtained through virtual screening （pIC50 ranged from 1.074 to 1.185， and Dcoking-score of protein-molecule interaction ranged from －9.730 to －7.467）. Pro268， Asp267， Gln171， Phe121 and Glu212 may be the key amino acids for the interaction between eEF2K inhibitors and target proteins， including hydrogen bonds， salt bridges and hydrophobicity. These 9 molecules are expected to be the lead compounds for the development of eEF2K inhibitors.

Objective To detect the influence of the first operation standardized or not on reoperation for differentiated thyroid carcinoma.Methods Retrospective analysis was conducted of the clinical data of 217 reoperation case of differentiated thyroid carcinoma from May 2009 to March 2018 in the 960th Hospital of the PLA Joint Logistic Support Force,including 58 male cases and 159 female cases,with the average age of 46.65 years (range from 19 to 76).According to the first operation standardized or not,all patientswas divided into standardized group (n =114) and non-standard group (n =103).Between the two groups,the number of dissected and metastatic lymph nodes,tumor pathology,recurrence range of glandular and central lymph nodes,number of lymph nodes removed and transferred,operation and drainage time,tumor invaded surrounding tissues,invasion sites,and complications were conducted.Results The overall gland recurrence rate was 20.2％ of 217 cases (44/217),8.8％ (10/114) in the standardized group and 33％ (34/103)in the non-standard group.The central group had a total recurrence rate of 38.7％ (84/217),and the standardized group and non-standard group were 22.8％ (26/114) and 56.3 ％ (58/103),respectively.For patients,the total cervical lymph node recurrence rate was 74.7％ (162/217),and the standardized group and non-standard group were 87.7％ (100/114),and 60.3％ (62/103),respectively.All of the difference was statistically significant(P =0.000,P =0.000,P =0.000).The operation time and drainage time of the standardized surgery group were significantly shorter than the non-standard group[(2.52±0.80)h vs(3.14±0.83) h,P=0.000;(4.37±1.28)d vs (5.16±1.41)d,P=0.000].Conclusions For differentiated thyroid carcinoma,the nonstandard treatment significantly affected the tumor residual rate in gland and lymph node metastasis rate of reoperation,andstandardized surgical treatmentshould be advocated.