Objective:To investigate the incidence and prothrombotic risk factors of postoperative venous thromboembolism(VTE) in Cushing′s disease and to further develop an assessment model to identify those at high risk of postoperative VTE events.Methods:A retrospective study was performed in 82 patients who were admitted to Huashan Hospital, Fudan University during January 2019 and January 2020 and diagnosed with Cushing′s disease. These patients underwent the evaluation about their clinical, hormonal, and coagulation parameters, as well as ultrasonography and pulmonary angio-CT when necessary. The least absolute shrinkage and selection operator(LASSO) regression analysis was used to screen independent risk factors, and a nomogram model for postsurgical VTE risk assessment in Cushing′s disease was initially established, and Bootstrap method was used for internal verification. Finally, the predictive model was evaluated for calibration and clinical applicability in the study cohort.Results:Nineteen patients(23.17%) developed VTE events, with 14 cases occurring after endoscopic transsphenoidal surgery. Compared to patients without VTE, those in the VTE group were older( P<0.001), had longer postoperative bed rest, higher rates of current infection, higher HbA 1C levels, and more severe glucose tolerance impairment(all P<0.05). Through LASSO regression analysis, two independent risk factors for postoperative VTE were identified: Age and current infection. Then a VTE risk assessment nomogram model was established to predict the patients at high risk of VTE. In the nomogram model for VTE risk assessment, the area under the receiver operating characteristic curve was 0.868(95% CI 0.787-0.949), with the calibration curve closely aligning with the ideal diagonal line and the clinical decision curve exceeding the two extreme curves. Conclusions:Advanced perioperative assessment needs to be taken to screen those with high VTE risks in patients diagnosed with Cushing′s disease. Additionally, during the perioperative period, patients with Cushing′s disease should undergo mandatory physical activity or prophylactic anticoagulant therapy.

Objective:A strong antithrombotic protein component， named PvQ， was purified and enriched from total protein of Pheretima vulgaris， a traditional Chinese medicine. Moreover， we evaluated its fibrinolytic and anticoagulant activity， and expected to provide reference for the research on antithrombotic substances of Pheretima. Method:A rapid in vitro activity-oriented separation combined with the AKTA-Pure protein purification system conducted on P. vulgaris. Meanwhile， the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time （Fibg-TT） method. And the in vitro thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature. Result:The PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of in vitro thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition， the changes of temperature and pH had significant effects on antithrombotic activity， and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions （pH<7）. While， the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions. Conclusion:A feasible preparation method of PvQ is established， and it can affect fibrin and fibrinogen at the same time， thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.

OBJECTIVE: To explore the regulation effect of myeloid leukemia No.1 Chinese herb medicine prescription combined with chemotherapy on Th17 cells in bone marrow fluid of AML patients, so as to provide guidance for improving AML treatment effect and patients' long-term survival. METHODS: Seventy patients with AML who were hospitalized in Department of Hematology, Wuwei People's Hospital from April 2017 to August 2019 were selected and enrolled in AML group, 25 healthy volunteers were selected and enrolled in control group; then according to therapeutic regimen, AML patients were divided into 2 groups: combined therapy group (myeloid leukemia NO.1 Chinese herb medicine prescription combined with chemotherapy) and non-combined therapy group (chemotherapy alone). Flow cytometry was used to detect the ratio of CD3 RESULTS: The ratio of CD3 CONCLUSION Th17 cells expression in bone marrow of newly diagnoses and relapsed AML patients significantly increase, and decrease significantly after treatment. Myeloid leukemia No.1 Chinese herb prescription combined with chemotherapy can significantly increase the CR rate and reduce the RL rate for AML.
Bone Marrow ; China ; Humans ; Leukemia, Myeloid, Acute/drug therapy* ; Medicine ; Prescriptions ; Th17 Cells ; Vascular Endothelial Growth Factor A

OBJECTIVE: To detect the level of vascular endothelial growth factor (VEGF) in bone marrow of patients with non-M3 acute leukemia (AL), and estimate its relationship with prognosis. METHODS: From January 2016 to December 2019, 114 patients with AL in department of Hematology, Wuwei People's Hospital were selected as study group, and 25 healthy volunteers were enrolled as control group. The concentration of VEGF in bone marrow was detected by ELISA. The patients were divided into high and low concentration group according to the level of VEGF. The overall survival (OS) and event-free survival (EFS) were compared among different groups. RESULTS: The level of VEGF in patients with AL was significantly higher than that in the control group. The median OS and EFS in the low concentration group was 34.5 and 32 months, respectively, while, in the high concentration group was 30 and 26 months, respectively. The differences between the two groups were statistically significant (P=0.010). There were significant differences in OS rate (P=0.035) and EFS rate (P=0.026) between low and high concentration group. Multivariate analysis showed that high VEGF concentration was an independent risk factor affecting OS (HR=2.619, 95%CI 1.070-6.406, P=0.035) and EFS (HR=2.221, 95%CI 1.074-4.552, P=0.031) in AL patients. CONCLUSION VEGF highly expresses in the bone marrow of patients with AL at initial diagnosis and relapse, and shows adverse effects on the prognosis.
Bone Marrow ; Disease-Free Survival ; Humans ; Leukemia, Myeloid, Acute ; Prognosis ; Vascular Endothelial Growth Factor A

Objective: To observe the incidence and treatment of radiation rectal injury complicated with anxiety, depression and somatic symptom disorder. Methods: A cross-sectional survey research method was carried out. Patients with radiation rectal injury managed by members of the editorial board of Chinese Journal of Gastrointestinal Surgery were the subjects of investigation. The inclusion criteria of the survey subjects: (1) patients suffered from pelvic tumors and received pelvic radiotherapy; (2) colonoscopy showed inflammatory reaction or ulcer in the rectum. Exclusion criteria: (1) patient had a history of psycho-somatic disease before radiotherapy; (2) patient was unable to use a smart phone, unable to read and understand the questions in the questionnaire displayed on the phone; (3) patient refused to sign an informed consent form. According to the SOMA self-rating scale, PHQ-15 self-rating scale, GAD-7 and PHQ-9 self-rating scale, the electronic questionnaire of "Psychological Survey of Radiation Proctitis" was designed. The questionnaire was sent to patients with radiation rectal injury managed by the committee through the WeChat group. Observational indicators: (1) radiation rectal injury symptom assessment: using SOMA self-rating scale, radiation rectal injury symptom classification: mild group (≤3 points), moderate group (4-6 points) and severe group (> 6 points); (2) incidence of anxiety, depression and physical disorder: using GAD-7, PHQ-9 and PHQ-15 self-rating scales respectively for assessment; (3) correlation of radiation rectal injury symptom grading with anxiety, depression, and somatic symptom disorder. Results: Seventy-one qualified questionnaires were collected, of which 41 (56.9%) were from Guangzhou. Among the 71 patients, 6 were males and 65 were females; the mean age was (55.7±9.3) years old and 48 patients (67.6%) were less than 60 years old; the median confirmed duration of radiation rectal injury was 2.0 (1.0, 5.0) years. (1) Evaluation of symptoms of radiation rectal injury: 18 cases of mild (25.4%), 27 cases of moderate (38.0%), and 26 cases of severe (36.6%). (2) Incidence of anxiety, depression and somatic disorder: 12 patients (16.9%) without comorbidities; 59 patients (83.1%) with anxiety, depression, or somatic disorder, of whom 2 patients only had anxiety, 1 patient only had depression, 9 only had somatic disorder, 2 had anxiety plus depression, 4 had anxiety plus somatic disorder, 2 had depression plus somatic disorder, and 40 had all three symptoms. (3) correlation of radiation rectal injury grading with anxiety, depression, and somatic symptom disorder: as compared to patients in mild group and moderate group, those in severe group had higher severity of anxiety and somatic symptom disorder (Z=-2.143, P=0.032; Z=-2.045, P=0.041), while there was no statistically significant difference of depression between mild group and moderate group (Z=-1.176, P=0.240). Pearson correlation analysis revealed that radiation rectal injury symptom score was positively correlated with anxiety (r=0.300, P=0.013), depression (r=0.287, P=0.015) and somatic symptom disorder (r=0.344, P=0.003). Conclusions: The incidence of anxiety, depression, and somatic symptom disorder in patients with radiation rectal injury is extremely high. It is necessary to strengthen the diagnosis and treatment of somatic symptom disorder, so as to alleviate the symptoms of patients with pelvic perineum pain and improve the quality of life.
Aged ; Anxiety ; Cross-Sectional Studies ; Depression ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Rectum ; Surveys and Questionnaires

Objective:To investigate whether human umbilical vein endothelial cell(HUVEC) vaccine combined with low dose docetaxel (DOC) could play a synergistic role in anti-breast cancer.Methods:BALB/c mice were randomly divided into normal saline group,HUVEC vaccine group,DOC group,and HUVEC vaccine combined with DOC treatment group (HUVEC-DOC) group.An experimental metastasis model by tail vein injection of EMT-6 breast cancer cells was employed to evaluate the anti-metastatic efficiency of the HUVEC-DOC combination treatment regime.Lymphocyte proliferation assay,cytotoxic T lymphocytes and an indirect enzyme-linked immunosorbent assay (ELISA) for detecting IFN-γ were used to investigate cellular immune responses elicited by the combination treatment regime.Results:Compared with HUVEC and DOC single drug group,the number of lung metastasis in HUVEC-DOC combination treatment group was significantly decreased(P<0.05).In vitro analysis of splenocytes isolated from immunized mice revealed an induction of cytotoxic T lymphocytes(CTLs) with a lytic activity against activated endothelium.IFN-γ in the serum of im-munized mice of the HUVEC-DOC combination treatment group was significantly higher than that in the other three groups(P<0.05). Conclusion:HUVEC vaccine with low dose of DOC could display synergistic anti-breast cancer effect.

Objective To investigate whether 2 repeats of mycobacterial microbial hot shock protein 70 (HSP70) 407-426 (2mHSP70407-426,M2) could act as an effective molecular adjuvant to enhance the anti-tumor efficiency of vascular endothelial growth factor (VEGF) fusion protein vaccine.Methods The gene encoding two tandem repeat sequences of HSP70407-426 was introduced to the terminus of the constructed hVEGF121 mutant 1 gene to prepare a recombined hVEGF121 mutant 2 protein vaccine by PCR technique.The anti-tumor efficacy of hVEGF121 mutant 2 protein vaccine was investigated using a H22 liver cancer subcutaneous tumor model.The 24 mice were randomly divided into three groups:model group,experiment 1 group (80 μg hVEGF121 mutant 1 protein) and experiment 2 group (80 μg hVEGF121 mutant 2 protein).The humoral and cellular immune responses were detected by ELISA and splenic lymphocyte proliferation assay.The anti-angiogenesis effect was evaluated by an intradermal tumor model.Results The tumor weight of the experimental 1 group and the experimental 2 group were (1.17± 0.25) g and (1.58 ± 0.28) g.Compared with the experimental 1 group,the tumor weight of tumor bearing mice in the experimental 2 group was lower significanfly(P ＜0.05).The anti-VEGF antibodies of the experimental 1 group and experimental 2 group was 0.54 ± 0.09,0.74 ± 0.1.Compared with experimental 1 group,higher titers of anti-VEGF antibody were detected in immune serum of the experimental 2 group with statistically significant(P ＜0.05).The proliferative activity of spleen lymphocytes in the experimental 1 group and the experimental 2 group was 0.26 ±0.03,0.36 ±0.04.Compared with the experimental 1 group,the vaccine immunization in experimental 2 group can stimulate the proliferation of spleen lymphocytes more effectively with statistically significant (P ＜ 0.05).Conclusion M2 could act as an effective adjuvant to help VEGF fusion protein vaccine to elicit a strong anti-tumor efficacy.

OBJECTIVETo observe the effects of low-intensity pulsed ultrasound (LIPUS) on repairing extracellular matrix in rabbit knee osteoarthritis and analyze its mechanism.

METHODSSixty adult female rabbits with an average weight of (2.0 ± 0.2) kg, were divided randomly into two groups (experimental group and control group, 30 rabbits in each group). All rabbits were replicated in right knees by Hulth method for knee osteoarthritis model. Two weeks after operation, the rabbits in experimental group were treated with LIPUS, and the ultrasonic frequency was (800 ± 5%)KHz and the maximum intensities of spatially averaged and time averaged (SATA) was (50 ± 10%) mw/cm2, for 1 time a day and every time 20 min, while the rabbits in control group were treated with sham LIPUS,the same operation with experimental group but without energy output. At the 2, 4, 8 weeks after treatment, 10 rabbits in each group were randomly killed for each time. The general changes of cartilage and its histopathological changes by HE staining were observed; the expression of collagen type II, proteoglycan, MMP-3, 7, 13 in cartilage were analyzed by immunohistochemical and RT-PCR technique; and the expression of NO in cartilage was analyzed by nitrate reduction method.

RESULTSOn the same observed time point, the damage degree of cartilage in experimental group was slighter than that of control group (P < 0.01), the expression of MMP-3, 7, 13 and NO in cartilage in experimental group was lower than that of control group (P < 0.01) while collagen type II and proteoglycan was higher than that of control group (P < 0.01).

CONCLUSIONLow-intensity pulsed ultrasound can repair the damaged cartilage by reducing the expression of MMP-3, 7, 13, inhibiting the secretion of NO and promoting the synthesis of collagen type II and proteoglycan in cartilage.

Animals ; Cartilage, Articular ; pathology ; Collagen Type II ; biosynthesis ; Extracellular Matrix ; metabolism ; Female ; Matrix Metalloproteinases ; analysis ; Nitric Oxide ; biosynthesis ; Osteoarthritis, Knee ; metabolism ; therapy ; Rabbits ; Ultrasonic Therapy ; methods

Objective:To observe the effects of low intensity pulsed ultrasound (LIPUS) on repairing extracellular matrix in rabbit knee osteoarthritis and analyze its mechanism. Methods:Sixty adult female rabbits with an average weight of (2.0± 0.2) kg,were divided randomly into two groups (experimental group and control group,30 rabbits in each group). All rabbits were replicated in right knees by Hulth method for knee osteoarthritis model. Two weeks after operation ,the rabbits in experi-mental group were treated with LIPUS,and the ultrasonic frequency was (800±5%)KHz and the maximum intensities of spa-tially averaged and time averaged(SATA) was (50±10%) mw/cm2,for 1 time a day and every time 20 min,while the rabbits in control group were treated with sham LIPUS ,the same operation with experimental group but without energy output. At the 2,4,8 weeks after treatment,10 rabbits in each group were randomly killed for each time. The general changes of cartilage and its histopathological changes by HE staining were observed;the expression of collagen typeⅡ,proteoglycan,MMP-3,7,13 in cartilage were analyzed by immunohistochemical and RT-PCR technique;and the expression of NO in cartilage was analyzed by nitrate reduction method. Results:On the same observed time point,the damage degree of cartilage in experimental group was slighter than that of control group(P<0.01),the expression of MMP-3,7,13 and NO in cartilage in experimental group was lower than that of control group (P<0.01) while collagen typeⅡand proteoglycan was higher than that of control group (P<0.01). Conclusion:Low intensity pulsed ultrasound can repair the damaged cartilage by reducing the expression of MMP-3,7,13,inhibiting the secretion of NO and promoting the synthesis of collagen typeⅡand proteoglycan in cartilage.

BACKGROUNDRosiglitazone is known as the most potent and specific peroxisome proliferators-activated receptor gamma (PPAR-gamma) ligand. It has potentially far-reaching effects on pathophysiological processes, from cancer to atherosclerosis and diabetes. However, it is not clear whether rosiglitazone affects the protein expression of transforming growth factor beta3 (TGF-beta3) and the cell proliferation in human uterine leiomyoma cells in vitro.

METHODSHuman uterine leiomyoma tissues were dissected and cultured. Cells were divided into 5 groups: one control group and other four groups with different concentrations of rosiglitazone (10(-7), 10(-8), 10(-9) and 10(-10) mol/L). Cells were cultured for 72 hours in serum-free Dulbecco's modified Eagle's medium. MTT reduction assay was used to detect the cell proliferation. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of PPAR-gamma and TGF-beta3. Immunofluorescence staining was used to detect the expressions of PPAR-gamma and TGF-beta3 proteins.

RESULTSMTT reduction assay indicated that the treatment with rosiglitazone (from 10(-7) to 10(-9) mol/L) resulted in an inhibition of the cell growths after 72 hours (P < 0.01). RT-PCR analysis revealed that 10(-7) mol/L rosiglitazone significantly affected the gene expression at 72-hour: PPAR-gamma mRNA expression was up-regulated and TGF-beta3 mRNA was down-regulated and rosiglitazone at the concentration of 10(-7) mol/L affected these most effectively (P < 0.01). Immunofluorescence staining demonstrated that treatment with 10(-7) mol/L rosiglitazone resulted in the significant changes of PPAR-gamma and TGF-beta3 protein expressions compared with the other treatment groups and the control group at 72-hour (P < 0.01). All the effects of rosiglitazone on uterine leiomyoma cells were dose- and time-dependent in vitro.

CONCLUSIONSThe present study demonstrates that the PPAR-gamma activator, rosiglitazone, inhibits the cell proliferation partly through the regulations of PPAR-gamma and TGF-beta3 expressions. The cross-talk between the signal pathways of PPAR-gamma and TGF-beta3 may be involved in the process.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Leiomyoma ; drug therapy ; pathology ; PPAR gamma ; agonists ; analysis ; genetics ; RNA, Messenger ; analysis ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta3 ; analysis ; genetics ; Uterine Neoplasms ; drug therapy ; pathology