Background and purpose:The ovarian mucinous tumor is one of the major subtypes of the ovari-an epithelial cancer. Ubiquitination is one of the main post-translational modifications, which has proven to be involved in tumorigenicity. Deubiquitinase is the protein enzyme that could reverse the process of ubiquitination to affect the initiation and progression of malignancies. This study aimed to analyze the expression and clinical application of deubiquitinase OTUB1 (OTU deubiquitinase, ubiquitin aldehyde binding 1) in ovarian primary mucinous tumors. Methods:This study collected 90 cases of ovarian primary mucinous tumors during 2010-2015 in Obstetrics and Gynecology Hospital of Fudan University, and then collected the clinicopathological information and performed the immunochemistry.Results:Fourteen out of 90 cases were ovarian primary mucinous cystadenoma, 17 were borderline mucinous tumor (intestinal type and intraepithelial carcinoma), and 59 were ovarian primary mucinous carcinoma. The expression rate and intensity of OTUB1 were much higher in malignant cases than those in benign ones (P<0.05). The expression rate and intensity of OTUB1 were much higher in cases with mucinous intraepithelial carcinoma than those in cases with intestinal type borderline mucinous tumor (P<0.05). The expression rate and intensity of OTUB1 increased with the advance of FIGO staging (P<0.05). The expression rate and intensity of OTUB1 were much higher in cases with involved fallopian tubes than those in cases without involved fallopian tubes (P<0.05). The expression rate and intensity of OTUB1were much higher in cases with involved uterus and omentum than those in cases with-out involvement (P<0.05). The expression rate and intensity of OTUB1were much higher in cases with lymph node metastasis than those in cases without involvement (P<0.05).Conclusion:There is significant difference in OTUB1 expression between ovarian primary mucinous carcinoma and benign mucinous cystadenoma. It is highly correlated to FIGO staging and invasion and metastasis of tumor. OTUB1 could be used in differential diagnosis and in monitoring the tumor initiation and progression in ovarian mucinous carcinoma.

Objective: To examine the correlation between exposure to polycyclic aromatic hydrocarbons ( PAHs ) and placental mitochondrial DNA ( mtDNA ) copy number among pregnant women, so as to provide the evidence for evaluation of potential effects of PAHs exposure during pregnancy on offspring health. Methods: A total of 200 pregnant women delivered at a tertiary hospital in Urumqi City during the period from January to October 2019, and their newborns were recruited, and grouped according to the time of delivery, including the heating group [delivery during the heating period ( from January to April ) ] and the non-heating group [delivery during the non-heating period (from July to October) ]. Subjects'age, ethnicity, educational level and type of home heating were collected, and the total concentration of 16 PAHs was determined in the blood samples of pregnant women and their babies using gas chromatography-mass spectrometry. Placental DNA was extracted, and placental mtDNA copy number was measured using real-time fluorescence quantitative PCR assay. In addition, the correlation between PAHs concentration and placental mtDNA copy number was examined using the Spearman rank correlation analysis. Results: There were 100 subjects in the heating group, which had a median age of 29 ( interquartile range, 3 ) years and had a mean gestational age of ( 275.06±0.72 ) days, and there were 100 subjects in the non-heating group, which had a median age of 29 ( interquartile range, 4 ) years and had a mean gestational age of ( 276.82±0.66 ) days. The total concentration of PAHs in the blood of pregnant women [15.71 (4.30) vs. 12.98 (5.49) μg/L; P<0.05 ], the total concentration of PAHs in neonatal blood [ 14.29 (4.25) vs. 11.24 (5.09) μg/L; P<0.05 ] and the placental mtDNA copy number [4.67 (1.18) vs. 4.51 (0.62); P<0.05] were all higher in the heating group than in the non-heating group. Spearman rank correlation analysis revealed that the total concentration of PAHs in the blood of pregnant women and neonates was positively correlated with placental mtDNA copy number ( rs=0.240, P=0.001; rs=0.273, P<0.001 ), and the total concentration of PAHs in the blood of pregnant women was positively correlated with the placental mtDNA copy number in the heating group ( rs=0.245, P=0.014 ), while the PAHs concentration in the neonatal blood was positively correlated with the placental mtDNA copy number in the non-heating group ( rs=0.292, P=0.003 ). Conclusions Exposure to PAHs positively correlates with placental mtDNA copy number among pregnant women, and there is a correlation between maternal exposure to PAHs and neonatal oxidative stress.

BACKGROUND: BuCy2 composed of large-dose oral busulfanum and cyclophosphamide has been one of the main conditioning regimens before transplantation, but oral busulfanum can result in many side effects.OBJECTIVE: To evaluate the efficacy and safety of the conditioning regimen with intravenous busulfanum (Ⅳ Bu) in hematopoietic stem cell transplantation (HSCT) for malignant hematopathy.DESIGN: Case observation.SETTING: Department of Hematology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 13 patients with chronic myelogenous leukemia (CML) and 6 patients with acute leukemia,who received allogeneic HSCT with the conditioning therapy of modified BuCy2, were enrolled at Institute of Hematopathy, Union Hospital, Huazhong University of Science and Technology from May to December 2006. There were 12 males and 7 females aged 14-50 years with an average of 33 years, and there were 9 cases of related transplantation and 10 cases of unrelated transplantation. Hematopoietic stem cells (HSCs) were harvested from peripheral blood of 18 subjects and bone marrow of 1 subject. All patients and their family members signed the informed consent.METHODS: All patients were treated with allogeneic HSCT with the conditioning therapy of intravenous BuCy2.All patients received the modified combination of busulfanum and cyclophosphamide that was commonly utilized currently.Hydroxycarbamide 40 mg/kg was orally taken ten days before transplantation. Arabinosylcytosin (Ara-C) 2 g/m2 was injected via vein nine days before transplantation. Busulfanum ampule 0.8 mg/kg (equal to 1 mg/kg oral administration)was given by central venous cannula eight, seven and six days before transplantation. The injection was performed over 2 hours controlled by infusion pump, once every 6 hours, totally 12 times. Busulfanum was diluted by saline or 5% glucose to about 0.5 g/L (about 10 times) before Ⅳ Bu. Cyclophosphamide 1.8 g/m2 was intravenously injected every day from five and four days before transplantation. Methyl-CCNU 250 mg/m2 was taken orally three days before transplantation. Occurrence of hepatic vano-occlusive disease (HVOD) and its side effects were determined. Follow-up was performed at month 6.5 after operation to observe disease recurrence and patient survival.MAIN OUTCOME MEASURES: Occurrence and side effects of HVOD 100 days after transplantation, and results of follow-up.RESULTS: Totally 19 patients were involved in the result analysis. In 13 days after transplantation, blood conversion,chromosome karyotype and DNA polymorphism tests verified that reconstruction of hematopoietic function was obtained and two patients got recurrence, of which one patient got complete remission after receiving fludarabine plus cytarabine plus granulocyte colony-stimulating factor (G-CSF) and HSCs from peripheral blood. DNA polymorhism test verified that re-engraftment was obtained. Another patient received chemotherapy. Other patients lived well.

OBJECTIVE To explore the safe and effective dose of sirolimus (Rapamycin,Sir) and its effect on seizure comorbidities. METHODS Immature C57BL/6 mice at postnatal 10 d of age were administered with kainic acid(KA) 12.0 mg · kg-1 intraperitoneally by a single injection to induce acute seizure. Sir 0.3, 1.0 and 3.0 mg · kg-1 was injected 24 h after seizure every other day until 3 d, 1 week, 3 weeks, 5 weeks and 6 weeks. Western blotting analysis was used to detect the expression and phos?phorylation level of S6 protein and to determine the minimum effective dose of Sir. Effect of the mini?mum effective dose of Sir on cognitive function and body growth was observed by several evaluations. Immunofluorescent intensity of Doublecortin (DCX) immunofluorescent staining was conducted to evaluate the development of neurons in the hippocampus. Morris water maze was used to assess the cognitive function. Tail suspension test, O maze and new object recognition test were used to study the anxiety-like behaviors of mice. RESULTS The result of Western blotting showed that Sir 0.3 mg · kg-1 had no significant effect on the phosphorylation of S6 protein in normal mice or KA mice, whereas 1.0 and 3.0 mg · kg- 1 could significantly inhibit the phosphorylation of S6 protein in KA mice (P<0.05). Sir 1.0 mg·kg-1 had no obvious effect on DCX-positive cells or body wass. Morris water maze showed that KA-induced seizure resulted in prolonged escape latency and swimming length (P<0.05), and a decreased crossing number of target quadrant (P<0.05). Sir 1.0 mg·kg-1 significantly reversed the deficit of cognitive function of KA-induced seizure mice (P<0.05), whereas no significant difference was found between Sir group and normal control group. Compared with normal control group, model group showed increased freezing time in tail suspension test (P<0.05), decreased migration length and reten?tion time in open arms in O maze (P<0.05), decreased retention time and touch frequency with new objects, migration length and average speed in new object recognition test (P<0.05). Sir 1.0 mg · kg-1 significantly reversed the above anxiety and depression status, whereas no significant difference was found between sirolimus group and normal control group. CONCLUSION Sir 1.0 mg · kg-1 inhibits the abnormal activation of mTOR pathway and the formation of epilepsy comorbidity in immature mice. Along with its mild side effect in development, Sir 1.0 mg · kg-1 will be an ideal dose to be used in the treatment of seizure in immature mice.

SIRT3, SIRT4 and SIRT5 are located in mitochondria and also known as mitochondrial sirtuins. They play important roles in regulating many cellular functions including cell survival, cell cycle or apoptosis, DNA repair and metabolism. Mitochondrial sirtuins are involved in the protection of mitochondrial integrity and energy metabolism under stress regulating the expression of neurotransmitter receptors, neurotrophins, extracellular matrix proteins and various transcription factors, thus involved in epileptogenesis triggered by both genetic or acquired factors. Here we review research progress on the actions of mitochondrial sirtuin in epilepsy; and discuss the challenges and perspectives of mitochondrial sirtuin as a potential therapeutic target for epilepsy.
Apoptosis ; Epilepsy/genetics* ; Humans ; Mitochondria/genetics* ; Sirtuin 3 ; Sirtuins

AIM: To investigate the biological characterics of human second-trimester fetal cord blood mesenchymal stem cells (MSC) and its application prospects in utero gene transfer/therapy (IUGT). METHODS: Nuclear cells separated from cord blood were cultured in DMEM medium. Surface antigens of the MSC were analyzed by the FACScan flow cytometry. Adipogenic and osteogenic mediums were used to assess the differentiation ability of the cells. Adenovirus vector deliver green fluorescent protein gene (Ad-GFP) was used to transfected the MSC and the expressing of GFP was detected by fluorescent microscope. The MSC were injected into the liver of newborn rat. The immunofluorescence analysis was conducted to determine the presence of double-positive CD105+/CD166+ cells in different organs of rats. MSC were subcutaneous injected into the human-nonobese diabetes/severe combined immunodeficiency disease (NOD/SCID) mice and carcinogenesises of the MSC in vivo were detected by pathological diagnosis. RESULTS: MSC could be separated from fetal cord blood. These cells were uniformly positive for CD29, CD44, CD59, CD105, CD166 and negative for CD34, CD45, CD80, CD86, HLA-DR. The cells had the abilities to differentiate into adipogenic and osteogenic cells in vitro, expressed the GFP at high levels (56 32%?3 28%). The MSC were located at different organs after injected into the newborn rats and didn't have carcinogenicity in vivo. CONCLUSION: Human second-trimester fetal cord blood MSC is an promising target cells in fetal IUGT.

Objective To explore the feasibility and clinical outcome of lateral cervical incision via sternocleidomastoid intermuscular approach(SMIA)in the treatment of primary hyperparathyroidism.Methods The clinical data of 64 patients with primary hyperparathyroidism who underwent unilateral parathyroid surgery in the First Affiliated Hospital,School of Medicine of Zhejiang University from January 2019 to June 2022 were retrospectively analyzed.They were divided into lateral cervical incision via sternocleidomastoid intermuscular approach group(SMIA group)and linea alba cervicalis approach group(LACA group)based on the surgical incision and access route.The differences in clinical features,surgery-related outcomes and postoperative functions of the anterior cervical region were compared between the two groups.The EQ-5D-5L scale was used to assess the subjective feeling of postoperative neck discomfort,while the Hollander Wound Assessment Scale was used to assess the clinical outcome of incision healing.Results There were no statistical differences between the two groups of patients in terms of age,gender,intraoperative bleeding,parathyroid hormone or blood calcium levels before and after surgery(P＞0.05).The duration of surgery was significantly shorter in the SMIA group than in the LACA group[(39.77±5.69)min vs.(54.41±4.66)min].There was a statistical difference between the two groups in functional protection of the anterior cervical region at 1 month and 12 months after surgery(1 month,84.67±3.74 vs.79.47±5.38,P＜0.001;12 months,93.80±2.52 vs.89.94±2.39,P＜0.001),and the SMIA group was better than the LACA group.The Hollander Incision Assessment Scale scores of the SMIA group were better than those of the LACA group at 6 months and 12 months after surgery,and the difference was statistically significant(6 months,1.93±0.58 vs.2.41±0.66,P=0.003;12 months,1.03±0.67 vs.1.74±0.62,P＜0.001).Conclusion Parathyroidectomy via sternocleidomastoid intermuscular approach through lateral cervical incision is a simple,safe and effective surgical procedure,which makes it easier to search for parathyroid lesions and shortens the surgical time compared with the traditional incision,and has obvious advantages in the protection of anterior cervical region function.

As a powerful tool to advance drug discovery, molecular imaging may provide new insights into the process of drug effect and therapy at cellular and molecular levels. When compared with other detection methods, fluorescence-based strategies are highly attractive and can be used to illuminate pathways of drugs' transport, with multi-color capacity, high specificity and good sensitivity. The conjugates of fluorescent molecules and therapeutic agents create exciting avenues for real-time monitoring of drug delivery and distribution, both in vitro and in vivo. In this short review, we discuss recent developments of small molecule-based fluorophore-drug conjugates, including non-cleavable and cleavable ones, that are capable of visualizing drug delivery.

To investigate the effect of a novel EZH2 inhibitor GSK126 on cell growth, apoptosis and migration of prostate cancer cells.Prostate cancer PC-3 and DU145 cells were treated with GSK126 at different doses. Cell growth was detected by sulforhodamine assay. Cell apoptosis was assayed by Annexin V-/PI kit. Transwell chamber and wound healing assays were conducted to detect cell migration. The mRNA level was detected by quantitative PCR, and protein expression was detected by Western blot analysis.GSK126 showed significant effect on cell growth and apoptosis when the dose was higher than 50 μmol/L. Wound healing assay revealed that scratch space in PC-3 cells was significantly increased in a dose-dependent manner in GSK126-treated groups[(247.2±24.4),(347.2±19.2) and (410.5±18.1) μm in low, medium and high dose (5.0, 20.0, 50.0 μmol/L), respectively] as compared with the control group[(171.3±17.8) μm](all<0.05). Transwell assay showed that migrated PC-3 cells in control group was 322.0±17.9,while those in GSK126-treated groups were 198.3±15.4 (low),82.7±6.2 (medium) and 30.2±4.1 (high), and the differences between the control group and GSK126-treated groups were significant(all<0.05). In addition, GSK126 up-regulated E-cadherin mRNA expression and down-regulated N-cadherin and Vimentin mRNA expression, whereas had no significant effect on Snail, Fibronectin and VEGF-A mRNA expression. The protein expression of E-cadherin was elevated but VEGF-A protein did not change in GSK126-treated groups. Similar results were exhibited in DU145 cell.GSK126 can significantly inhibit cell migration and invasion in prostate cancer PC-3 and DU145 cells, which may be resulted from its effect on epithelial-mesenchymal transition. GSK126 may be used as a potential anti-prostate cancer dug in clinic.
Apoptosis ; drug effects ; Cadherins ; analysis ; drug effects ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Drug Screening Assays, Antitumor ; methods ; Enhancer of Zeste Homolog 2 Protein ; analysis ; drug effects ; metabolism ; Fibronectins ; analysis ; drug effects ; metabolism ; Humans ; Indoles ; pharmacology ; Male ; Prostatic Neoplasms ; chemistry ; genetics ; physiopathology ; Pyridones ; pharmacology ; RNA, Messenger ; Up-Regulation ; drug effects ; Vascular Endothelial Growth Factor A ; analysis ; drug effects ; Vimentin ; analysis ; drug effects ; metabolism

Objective:To explore the effect and mechanism of cytochrome P450 1A1 (CYP1A1) on regulating phagocytosis of macrophage treated with Escherichia coli ( E.coli). Methods:① The mouse leukemia cells lines of monocyte macrophage RAW264.7 (RAW) were cultured in vitro and treated with 30 multiplicity of infection (MOI) dosages of E.coli for 40 minutes, glycerin control group was set up to observe the change of CYP1A1 during infection. ② The RAW cells with CYP1A1 overexpression (CYP1A1/RAW) and knock out (CYP1A1 KO/RAW) were cultured in vitro and treated with 30 MOI E. coli for 40 minutes, while the negative controlled RAW cells (NC/RAW) were established as control to observe the relationship between cell phagocytosis and CYP1A1 expression, and the effect of CYP1A1 on phagocytic receptor [scavenger receptor-A (SR-A)] and its signal pathway [mitogen-activated protein kinase (MAPK) pathway]. ③ NC/RAW and CYP1A1 KO/RAW cells were cultured in vitro and pretreated with 1 μmol/L extracellular signal-regulated kinase (ERK) inhibitor (U0126) for 2 hours, and then treated with 30 MOI E.coli for 40 minutes, phosphate buffered solution (PBS) control group was set up to observe whether the effect of CYP1A1 on phagocytosis through controlled the MAPK pathway. ④ The RAW cells were cultured in vitro and pretreated with 100 nmol/L CYP1A1 hydroxylase active product 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] for 2 hours, and then treated with 30 MOI E.coli for 40 minutes, and PBS control group was set up to observe whether the effect of CYP1A1 on phagocytosis was related to CYP1A1 hydroxylating metabolite. ⑤ The RAW cells with overexpression CYP1A1 hydroxylase-activity mutation (CYP1A1m/RAW) were cultured in vitro and treated with 30 MOI E.coli for 40 minutes, the CYP1A1/RAW cells were set up as control group to observe whether the effect of CYP1A1 on phagocytosis was related to CYP1A1 hydroxylase-activity. Results:① Compared with glycerin control group, CYP1A1 mRNA expression was significantly increased by E.coli stimulation (2 -ΔΔCt: 7.79±0.71 vs. 1.00±0.00, P < 0.05), indicating that CYP1A1 might participate in regulating infection progress. ② Compared with NC/RAW cells, the number of E.coli colonies phagocytized by CYP1A1/RAW cells was significantly decreased after 40 minutes of E.coli stimulation (×10 3 CFU/mL: 4.67±3.06 vs. 15.67±5.03, P < 0.05), while CYP1A1 KO/RAW cells had a significant increase in the number of E.coli colonies phagocytized (×10 3 CFU/mL: 46.00±5.29 vs. 15.67±5.03, P < 0.05), suggesting that CYP1A1 might negatively control macrophage phagocytosis function. Meanwhile, compared with NC/RAW cells, the expression of SR-A mRNA in CYP1A1/RAW cells was significantly down-regulated (2 -ΔΔCt: 0.31±0.03 vs. 1.00±0.00, P < 0.05), and the activation level of ERK was significantly reduced. However, the expression of SR-A mRNA in CYP1A1 KO/RAW cells was significantly up-regulated (2 -ΔΔCt: 3.74±0.25 vs. 1.00±0.00, P < 0.05), and the activation of ERK was enhanced, indicating that CYP1A1 could negatively regulate phagocytic receptors and their signaling pathways.③ Compared with PBS, U0126 pretreatment significantly inhibited the CYP1A1 knockout induced upregulation of SR-A mRNA expression (2 -ΔΔCt: 0.62±0.05 vs. 4.38±0.39, P < 0.05) and ERK activation, and inhibited the enhancement of phagocytosis in macrophages induced by CYP1A1 knock out [ E.coli colonies phagocytized by cells (×10 3 CFU/mL): 12.67±1.15 vs. 45.33±4.16, P < 0.05], suggesting that CYP1A1 inhibited macrophage phagocytosis function by regulating ERK activation. ④ Compared with PBS, the phagocytosis of RAW cells pretreated with 12(S)-HETE did not change significantly [ E.coli colonies phagocytized by cells (×10 3 CFU/mL): 17.00±1.00 vs. 16.33±2.52, P > 0.05], suggesting that CYP1A1 might not control phagocytosis function by its hydroxylase-activity metabolism 12(S)-HETE. ⑤ Compared with CYP1A1/RAW cells, there was no significant change in the phagocytic function of CYP1A1m/RAW cells [ E.coli colonies phagocytized by cells (×10 3 CFU/mL): 3.67±1.15 vs. 3.33±0.58, P > 0.05], suggesting that CYP1A1 might not control phagocytosis function by its hydroxylase-activity. Conclusion:CYP1A1 can negatively regulate the phagocytosis of macrophages by inhibiting the activation of ERK and reducing the expression of SR-A, but this regulatory effect is not related to the activity of CYP1A1 hydroxylase and its pro-inflammatory metabolism 12(S)-HETE.