Objective To evaluate the growth inhibition and apoptosis of human monocytic leukemia THP-1 cell line by using 5,8-dimethyl-2-β-hydroxyisovalerylshikonin (SK36) and explore its preliminary mechanism. Methods CCK colorimetric assay and cell counting was used to examine the growth inhibition of shikonin on THP-1 cells. The apoptosis of THP-1 cells was detected by Annexin V/PI double labeling. The activation of Caspase-3 apoptosis pathway was determined by FCM. The apoptosis and the necrosis of THP-1 cells were detected by the laser scanning confocal microscopy. Results When the THP-1 cells were treated with SK36 at 1.02 μg/ml for 24 h and 48 h, the growth inhibition was dose-dependent. The cell apoptotic rate of THP-1 cells treated with 1.02 μg/ml evaluated by FCM with Annexin V/PI double labeling staining were (40.61 ±2.13) % and (67.40±9.15) % at 24 h and 48 h after treatment, respectively, which were significantly higher than that of the control group [(16.97±0.61) %] ([ = 18.444, t = 9.528, P <0.01). SK36 could induce THP-1 cells apoptosis involving the activation of Caspase-3 (F= 323.61, P<0.01). Conclusion SK36 can induce human THP-1 cells to undergo apoptosis, and its primary mechanism was to activate the Caspase-3.

Objective To evaluate the clinical efficacy and adverse effects of CAG regimen in treatment of primary, refractory and relapsed acute myeloid leukemia (AML) and high risk myelodysplastic syndrome (MDS), and analyse the factors influencing long-term survival. Methods Sixty-one patients with AML ( primary, n = 27; refractory, n = 18; relapsed, n = 16) and 9 patients with MDS were treated with CAG regimen. Examinations on liver and renal function, electrocardiogram and bone marrow cytology were performed before and after treatment, and adverse effects of CAG were observed. Short-term efficacy was evaluated based on clinical manifestation, peripheral blood and bone marrow cytologic examinations. Patients were followed up, overall survival ( OS) and disease free survival ( DFS) were analysed, and long-term efficacy of CAG regimen was evaluated. The factors influencing long-term survival were analysed by Log-rank test of survival curve. Results After a course of treatment by CAG regimen, the total effective rate was 71% , and 34 patients (49%) experienced complete remission. The median time of follow up was 45 months, the median OS was 28 months, and the median DFS was 23 months. Age, level of lactate dehydrogenase (LDH), remission condition after a course of treatment by CAG regimen and adoption of HD-Ara-C regimen as consolidation treatment were influencing factors for OS and DFS. The dominant clinical adverse effects were bone marrow depression, with 13 d as the median duration of agranulocytosis ( neutrophil <0.5 ×10~9/L) and 9 d as the median duration of thrombocytopenia (platelet <20 ×10~9/L). Conclusion CAG regimen may lead to favourable therapeutic effects in treatment of primary, refractory and relapsed AML and high risk MDS, and may yield less adverse effects and better long-term therapeutic effects. Age, level of LDH, remission condition after a course of treatment and adoption of HD-Ara-C regimen as consolidation treatment are dominant influencing factors for survival.

Objective To analysis long-term effects and safety of arsenic trioxide (ATO)-based induction and maintenance therapy in newly diagnosed acute promyelocytic leukemia (APL). Methods Retrospective analysis induction remission and post-remission treatment of 62 newly diagnosed APL patients was performed. These cases were followed up for 5 and 7 years. Results The complete remission (CR) rate was similar in ATO/all-trans retinoic acid (ATRA) induction group and ATRA/chemotherapy induction group.However, the former group has the shorter time to CR. The negative rate of PML-RARα fusion gene after induction without ATO was less than that of ATO group (86.2 % vs 56.3 %, P ＜0.05). After CR, the 5-year overall survival (OS) between ATO-base rotation maintenance group and chemotherapy-base rotation maintenance group showed that the former was (94.4±5.4) %, the latter is (45.5±10.2) %; 7-year OS was (52.5±23.7) % and (27.3±9.3) %; 5-year disease free survivals (DFS) was (94.7±5,5) % and (41.3±10.1) %; 7-year DFS was (52.6±23.7) % and (27.5±9.4) %. There was significant different in 5-year or 7-year OS and DFS between two groups (P ＜0.05). The relapse rates of the two groups in post-remission treatment were 14.7 % and 37.0 % (P ＜0.05). Conclusion ATO combined ATRA induction therapy increased the negative rate of PML-RARα fusion gene. ATO-base rotation maintenance improved long-term outcome and decreased the rate of relapse. Furthermore, ATO appeared to be generally safe and well tolerated.

We transformed the fip-fve gene into Pichia pastoris GS115 for inducible and constitutive expression to obtain feasible bioactvie recombinant Fip-fve. The fip-fve gene was cloned from Flammulina velutipes fruting body by PCR and ligated to pPIC9 to construct inducible expression vector pPIC9-FIP-fve, and promotor pgap was used to replace the paox1 to construct constitutive expression vector pPIC9-PGAP-FIP-fve. These two vectors were used to transform P. pastoris by PEG method. The fip-fve was expressed after histamine-absence screening and yeast colony PCR. The inducible expression level reached 158.2 mg/L at the fourth day and the constitutive expression level was 46.3 mg/L and 29.5 mg/L using glucose and glycerol, respectively. The SDS-PAGE and Western blotting both proved the correctness of rFip-fve, and the hemagglutination test indicats the rFip-fve's bioactivity.
Electrophoresis, Polyacrylamide Gel ; Flammulina ; chemistry ; Fungal Proteins ; biosynthesis ; Genetic Vectors ; Pichia ; metabolism ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Recombinant Proteins ; biosynthesis

OBJECTIVETo establish a new method for the identification of Schisandra sphenanthera and S. chinensis.

METHODRandom amplified polymorphic DNA-Sequence characterized applied region (RAPD-SCAR) method was applied to screen primers.

RESULTScreening from 100 primers, only 2 random primers, which can be used to identify S. sphenanthera and S. chinensis accurately with a good reproducibility. It worked to fit them into sequence characterized applied region.

CONCLUSIONRAPD-SCAR can be used to identify S. sphenanthera and S. chinensis accurately.

Objective To obtain methamphetamine concentration profiles in saliva and urine samples of drug addicts and to screen the colloidal gold strip. Methods Methamphetamine concentration in saliva and urine samples of drug addicts was determined by liquid chromatography tandem mass spectrometry. The initial screening was obtained by colloidal gold strip test. The results were compared and analyzed. Results using the method of protein and fluid MRM scan method to detect direct precipitation, saliva is linear in the range of 1~100ng/mL, the linear correlation coefficient is 0.9987, the detection limit is 0.1ng/mL, the limit of quantification was 1ng/mL, the urine is linear in the range of 1~100ng/mL, the linear correlation coefficient is 0.9943, the detection limit is 0.5ng/mL, the limit of quantification was 1ng/mL. Saliva and urine samples diluted, the concentration in the linear range. Saliva and urine samples of four types of methamphetamine colloidal gold reagent strip were screened directly, and the results were judged visually. Conclusion the detection rate of colloidal gold strip is about 79%, the detection rate of saliva is about 81%, and the detection rate can be increased to more than 93% by using two reagent strips. Combined with the initial screening results and the instrument confirmation concentration, it can be found that the gray zone setting and sensitivity setting have certain influence on the detection rate, and it is suggested to improve the sensitivity to meet the needs of screening.

The components of the tumor microenvironment have a complex composition and play an important regulatory role in tumor evolution.The extracellular matrix(ECM)serves as a vital mediator in regulating cancer advancement within the TME.Incorporating ECM into tumor modeling allows for a more comprehensive simulation of the TME in vitro.As a medium for organoid growth,the matrix gel fulfills the functions of ECM.ECM is normally derived from the recombinant basement membrane of mouse sarcoma(Matrigel);however,batch-to-batch variations in Matrigel affect experiment reproducibility.To overcome such issues,researchers have designed synthetic matrix gels.This paper provides an overview of the current research in the application of matrix gels for tumor organoid modeling,offering insights for continuous optimization of organoid culture,thereby achieving more efficient and cost-effective organoid construction and advancing the translation of basic research to clinical applications.

AIM: To observe the effects of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO) and transforming growth factor-β1 (TGF-β1) of radiation-induced lung injury rats by Kiwi fruit essence (unsaturated fatty acid of actinidia chinesis planch seed oil). METHODS: According to random number table, 90 Sprague-Dawley rats were divided into the control group, model group, 60 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 120 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 240 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 18 animals were included in each group. Except for the control group, rats in the remaining groups were constructed by 6MV-X-ray 18Gy full chest radiation, one week before modeling, the rats in the last 3 groups were given (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil. The first two groups were given 0.9% sodium chloride solution by gavage, once a day in each rat. 14 days, 28 days, and 56 days after radiation, the rats were randomly sacrificed and their chests were cut, and ave lung tissue was saved. HE and Masson staining were performed to observe the pathological changes, and the content of SOD, GSH-Px, MPO was determined. The expression of TGF-β1 was analyzed by immunohistochemical staining. RESULTS: Compared with the model group, (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil significantly reduced the degree of lung alveolitis and radiation pulmonary fibrosis, reduced the content of hydroxyproline (HYP), MPO, increased the antioxidant enzymes SOD and GSH-Px content, down-regulated the expression of TGF-β1.There were significant differences in the above-mentioned indicators among the intervention groups of (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil group, and it was positively correlated with dosage. CONCLUSION: Unsaturated fatty acid of actinidia chinesis planch seed oil has a preventive effect on radiation-induced lung injury, and its mechanism may be related to the reduction of oxidative stress damage and down-regulation of TGF-β1 expression.

Objective    To investigate the correlation between lung ultrasonography and pulmonary complications after cardiac surgery. Methods    Fifty-two patients after cardiac surgery in our hospital from January to May 2017 were recruited. There were 27 males and 25 females, aged 60.50±10.43 years. Lung ultrasonography was performed by specially trained observers, video data were saved, and lung ultrasound score (LUS) were recorded. The correlation between the LUS and the patients' pulmonary function was evaluated. Results    LUS was 17.80±3.87, which was negatively correlated to the ratio of arterial PO2 to the inspired oxygen fraction (PaO2/FiO2) during examination, without significant difference (r=–0.363, P=0.095), but significantly negatively correlated to PaO2/FiO2 changes 24 hours postoperatively (r=–0.464, P=0.034). Conclusion    The changes of lung ventilation area may occur earlier than the changes of lung function. Bedside LUS is an effective method for clinical monitoring of pulmonary complications.

Objective:To investigate the epidemiological characteristics, diagnosis, treat-ment and prognosis of gallbladder cancer in China from 2010 to 2017.Methods:The single disease retrospective registration cohort study was conducted. Based on the concept of the real world study, the clinicopathological data, from multicenter retrospective clinical data database of gallbladder cancer of Chinese Research Group of Gallbladder Cancer (CRGGC), of 6 159 patients with gallbladder cancer who were admitted to 42 hospitals from January 2010 to December 2017 were collected. Observation indicators: (1) case resources; (2) age and sex distribution; (3) diagnosis; (4) surgical treatment and prognosis; (5) multimodality therapy and prognosis. The follow-up data of the 42 hospitals were collected and analyzed by the CRGGC. The main outcome indicator was the overall survival time from date of operation for surgical patients or date of diagnosis for non-surgical patients to the end of outcome event or the last follow-up. Measurement data with normal distribu-tion were represented as Mean±SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(range), and com-parison between groups was conducted using the U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Univariate analysis was performed using the Logistic forced regression model, and variables with P<0.1 in the univariate analysis were included for multivariate analysis. Multivariate analysis was performed using the Logistic stepwise regression model. The life table method was used to calculate survival rates and the Kaplan-Meier method was used to draw survival curves. Log-rank test was used for survival analysis. Results:(1) Case resources: of the 42 hospitals, there were 35 class A of tertiary hospitals and 7 class B of tertiary hospitals, 16 hospitals with high admission of gallbladder cancer and 26 hospitals with low admission of gallbladder cancer, respectively. Geographical distribution of the 42 hospitals: there were 9 hospitals in central China, 5 hospitals in northeast China, 22 hospitals in eastern China and 6 hospitals in western China. Geographical distribution of the 6 159 patients: there were 2 154 cases(34.973%) from central China, 705 cases(11.447%) from northeast China, 1 969 cases(31.969%) from eastern China and 1 331 cases(21.611%) from western China. The total average number of cases undergoing diagnosis and treatment in hospitals of the 6 159 patients was 18.3±4.5 per year, in which the average number of cases undergoing diagnosis and treatment in hospitals of 4 974 patients(80.760%) from hospitals with high admission of gallbladder cancer was 38.8±8.9 per year and the average number of cases undergoing diagnosis and treatment in hospitals of 1 185 patients(19.240%) from hospitals with low admission of gallbladder cancer was 5.7±1.9 per year. (2) Age and sex distribution: the age of 6 159 patients diagnosed as gallbladder cancer was 64(56,71) years, in which the age of 2 247 male patients(36.483%) diagnosed as gallbladder cancer was 64(58,71)years and the age of 3 912 female patients(63.517%) diagnosed as gallbladder cancer was 63(55,71)years. The sex ratio of female to male was 1.74:1. Of 6 159 patients, 3 886 cases(63.095%) were diagnosed as gallbladder cancer at 56 to 75 years old. There was a significant difference on age at diagnosis between male and female patients ( Z=-3.99, P<0.001). (3) Diagnosis: of 6 159 patients, 2 503 cases(40.640%) were initially diagnosed as gallbladder cancer and 3 656 cases(59.360%) were initially diagnosed as non-gallbladder cancer. There were 2 110 patients(34.259%) not undergoing surgical treatment, of which 200 cases(9.479%) were initially diagnosed as gallbladder cancer and 1 910 cases(90.521%) were initially diagnosed as non-gallbladder cancer. There were 4 049 patients(65.741%) undergoing surgical treatment, of which 2 303 cases(56.878%) were initially diagnosed as gallbladder cancer and 1 746 cases(43.122%) were initial diagnosed as non-gallbladder cancer. Of the 1 746 patients who were initially diagnosed as non-gallbladder cancer, there were 774 cases(19.116%) diagnosed as gallbladder cancer during operation and 972 cases(24.006%) diagnosed as gallbladder cancer after operation. Of 6 159 patients, there were 2 521 cases(40.932%), 2 335 cases(37.912%) and 1 114 cases(18.087%) undergoing ultrasound, computed tomography (CT) or magnetic resonance imaging (MRI) examination before initial diagnosis, respec-tively, and there were 3 259 cases(52.914%), 3 172 cases(51.502%) and 4 016 cases(65.205%) undergoing serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis, respectively. One patient may underwent multiple examinations. Results of univariate analysis showed that geographical distribution of hospitals (eastern China or western China), age ≥72 years, gallbladder cancer annual admission of hospitals, whether undergoing ultrasound, CT, MRI, serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis were related factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.45, 1.98, 0.69, 0.68, 2.43, 0.41, 1.63, 0.41, 0.39, 0.42, 95% confidence interval as 1.21-1.74, 1.64-2.40, 0.59-0.80, 0.60-0.78, 2.19-2.70, 0.37-0.45, 1.43-1.86, 0.37-0.45, 0.35-0.43, 0.38-0.47, P<0.05). Results of multivariate analysis showed that geographical distribution of hospitals (eastern China or western China), sex, age ≥72 years, gallbladder cancer annual admission of hospitals and cases undergoing ultrasound, CT, serum CA19-9 examination before initially diagnosis were indepen-dent influencing factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.36, 1.42, 0.89, 0.67, 1.85, 1.56, 1.57, 0.39, 95% confidence interval as 1.13-1.64, 1.16-1.73, 0.79-0.99, 0.57-0.78, 1.60-2.14, 1.38-1.77, 1.38-1.79, 0.35-0.43, P<0.05). (4) Surgical treatment and prognosis. Of the 4 049 patients undergoing surgical treatment, there were 2 447 cases(60.435%) with complete pathological staging data and follow-up data. Cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb were 85(3.474%), 201(8.214%), 71(2.902%), 890(36.371%), 382(15.611%), 33(1.348%) and 785(32.080%), respectively. The median follow-up time and median postoperative overall survival time of the 2 447 cases were 55.75 months (95% confidence interval as 52.78-58.35) and 23.46 months (95% confidence interval as 21.23-25.71), respectively. There was a significant difference in the overall survival between cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb ( χ2=512.47, P<0.001). Of the 4 049 patients undergoing surgical treatment, there were 2 988 cases(73.796%) with resectable tumor, 177 cases(4.371%) with unresectable tumor and 884 cases(21.833%) with tumor unassessable for resectabi-lity. Of the 2 988 cases with resectable tumor, there were 2 036 cases(68.139%) undergoing radical resection, 504 cases(16.867%) undergoing non-radical resection and 448 cases(14.994%) with operation unassessable for curative effect. Of the 2 447 cases with complete pathological staging data and follow-up data who underwent surgical treatment, there were 53 cases(2.166%) with unresectable tumor, 300 cases(12.260%) with resectable tumor and receiving non-radical resection, 1 441 cases(58.888%) with resectable tumor and receiving radical resection, 653 cases(26.686%) with resectable tumor and receiving operation unassessable for curative effect. There were 733 cases not undergoing surgical treatment with complete pathological staging data and follow-up data. There was a significant difference in the overall survival between cases not undergoing surgical treatment, cases undergoing surgical treatment for unresectable tumor, cases undergoing non-radical resection for resectable tumor and cases undergoing radical resection for resectable tumor ( χ2=121.04, P<0.001). (5) Multimodality therapy and prognosis: of 6 159 patients, there were 541 cases(8.784%) under-going postoperative adjuvant chemotherapy and advanced chemotherapy, 76 cases(1.234%) under-going radiotherapy. There were 1 170 advanced gallbladder cancer (pathological staging ≥stage Ⅲa) patients undergoing radical resection, including 126 cases(10.769%) with post-operative adjuvant chemotherapy and 1 044 cases(89.231%) without postoperative adjuvant chemo-therapy. There was no significant difference in the overall survival between cases with post-operative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.23, P=0.629). There were 658 patients with pathological staging as stage Ⅲa who underwent radical resection, including 66 cases(10.030%) with postoperative adjuvant chemotherapy and 592 cases(89.970%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.05, P=0.817). There were 512 patients with pathological staging ≥stage Ⅲb who underwent radical resection, including 60 cases(11.719%) with postoperative adjuvant chemotherapy and 452 cases(88.281%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemo-therapy and cases without post-operative adjuvant chemo-therapy ( χ2=1.50, P=0.220). Conclusions:There are more women than men with gallbladder cancer in China and more than half of patients are diagnosed at the age of 56 to 75 years. Cases undergoing ultrasound, CT, serum CA19-9 examination before initial diagnosis are independent influencing factors influencing initial diagnosis of gallbladder cancer patients. Preoperative resectability evaluation can improve the therapy strategy and patient prognosis. Adjuvant chemotherapy for gallbladder cancer is not standardized and in low proportion in China.