ABSTRACT
OBJECTIVE: This study aims at evaluating the effects of ciprofol on the induction and maintenance of general anesthesia in patients undergoing kidney transplantation. PATIENTS AND METHODS: This prospective, randomized, single-blind study enrolled 120 patients aged 18-65 years who underwent general anesthesia for kidney transplantation. The patients were randomized into a ciprofol group (group C) and a propofol group (group P). Anesthesia induction: group C had injected IV with ciprofol 0.4 mg/kg, group P had injected IV with propofol 2.0 mg/kg, while both groups had injected IV with sufentanil 0.4-0.5 µg/kg and cisatracurium 0.2 mg/kg. Anesthesia maintenance: ciprofol was injected IV with 0.8-2.4 mgâ¢kg-1â¢h-1 in group C, propofol was injected IV with 4-12 mgâ¢kg-1â¢h-1 in group P, while remifentanil was injected IV with 8-15 µgâ¢kg-1â¢h-1 and cisatracurium was injected IV with 0.1-0.2mgâ¢kg-1â¢h-1, with the bispectral index (BIS) maintained at 40-60 during the operation. RESULTS: The success rate of sedation in both groups was 100%. Compared with the P group, in group C the time of disappearance of the eyelash reflex and a decline in the BIS to 60 was shorter (p<0.001); the time of awakening was prolonged (p<0.001); the number of sedative drugs administered was reduced (p<0.001); MAP fluctuated less five mins after transplantation (p<0.01); the incidence of injection pain during induction was reduced (p<0.001) and intraoperative hypotension was decreased(p<0.01). CONCLUSIONS: Ciprofol is safe and effective for anesthesia induction and maintenance in kidney transplantation and its sedative effect is better than that of propofol.
Subject(s)
Kidney Transplantation , Propofol , Anesthesia, General/adverse effects , Anesthetics, Intravenous , Double-Blind Method , Humans , Prospective Studies , Single-Blind MethodABSTRACT
Objective: To summarize the etiological characteristics and clinical treatment effects of 17 patients with medication-related osteonecrosis of the jaw (MRONJ). Methods: The clinical data of 17 patients with MRONJ admitted to the Department of Oral and Maxillofacial Surgery, the First People's Hospital of Jinzhong, Shanxi Province, from July 2016 to December 2019 were retrospectively analyzed, including 9 males and 8 females, aged (63.6±9.6) years old (43-82 years old). Descriptive analysis of the primary disease, onset factors, site of disease, clinical manifestations, treatment methods, and treatment effects was conducted through follow-up for at least 1 year. Results: Among the primary diseases of the 17 cases, 12 were malignant tumors, and 5 were osteoporosis. There were 13 cases with a history of a trigger event (tooth extraction or unsuited removable denture). Six cases occurred in the maxilla, 10 cases occurred in the mandible, and 1 case involved both the upper and lower jaws. For the most common medication used, bisphosphonate was used in 16 cases including 5 cases with concomitant use of angiogenesis-inhibiting drugs. There was 1 case resulted from receptor activator of NF-κB ligand (RANKL) monoclonal antibody application. The duration of medication application was (10.1±3.9) months (3-18 months). All 17 cases were treated surgically. Totally 15 patients healed well after surgical treatment and the other 2 patients, who had poor soft tissue healing after surgery, healed well after a second operation. Conclusions: Tooth extraction might be a major trigger factor for the onset of MRONJ in the mandible. The disease was more possibly occured in the mandible than in the maxilla. Appropriate surgical treatment could achieve a good clinical outcome.
Subject(s)
Bisphosphonate-Associated Osteonecrosis of the Jaw , Bone Density Conservation Agents , Adult , Aged , Aged, 80 and over , Bisphosphonate-Associated Osteonecrosis of the Jaw/etiology , Bisphosphonate-Associated Osteonecrosis of the Jaw/therapy , Bone Density Conservation Agents/adverse effects , Diphosphonates , Female , Humans , Male , Mandible , Middle Aged , Retrospective StudiesABSTRACT
ABSTRACT: Objective To analyze the characteristics of the bilateral external ears of Uygur adults by directly observing the morphological characteristics of the external ears of Uygur adults and classifying each feature. The frequency distribution of the characteristics was calculated to provide reference for forensic identification. Methods The 210 cases ï¼75 males and 135 femalesï¼ of bilateral external ear photos of Uygur adults in Xinjiang that met the inclusion criteria were collected. The frequencies of the features of the external ear were recorded and distinguished between the two sexes and the different sides. The data were statistically analyzed by SPSS 21.0 statistical software. Results The shapes of the external ears of males and females were commonly oblique or rectangular ï¼34.67% of the left external ear of males and 41.33% of the right were oblique; 30.37% of the left and right external ear of females were rectangularï¼, while triangular ears were the rare variants and the least common. Sex and bilateral differences were observed as regards the form of the helix in the subjects. Normally rolled helix was the most common ï¼58.67% males and 61.48% females for the left ear; 60.00% males and 72.59% females for the right earï¼. Wide covering scapha helix was the most rare for the male left ear and flat helix was the most rare for the female right ear. Square and free earlobes were the most common ï¼49.33% males and 62.96% females for the left ear; 40.00% males and 54.81% females for the right earï¼, whereas triangular earlobes were rarely seen. Single knob tragus ï¼40.00% males and 37.78% females for the left ear; 37.33% males and 33.33% females for the right earï¼ and projection type of Darwin's tubercle ï¼50.67% males and 40.00% females for the left ear; 48.00% males and 39.26% females for the right earï¼ were found to be common. Conclusion The characteristics of the bilateral external ears of male and female Uygur adults have differences, which can be used for forensic identification.
Subject(s)
Ear, External , Sex Characteristics , Adult , Ethnicity , Female , Humans , MaleABSTRACT
OBJECTIVE: Lentivirus-delivered microRNA (miR) has been reported to improve survival outcomes and organ dysfunction. The present study is aimed to explore whether sepsis-associated miR, miR-181-5p, could mitigate sepsis-induced inflammation and organ injury by the lentivirus-expressing system. MATERIALS AND METHODS: Cecal ligation and puncture (CLP)-operated mice were treated with lentivirus-expressing miR-181-5p (miR-agomir) 7 days before surgical operation by intravenous injection. Acute renal and hepatic injuries were assessed using specific biomarkers. Survival outcomes were evaluated following CLP operation within 72 hours. RESULTS: Lentivirus-delivered miR-181-5p improves survival outcomes of CLP-induced septic mice. The rescue of miR-181-5p expression by lentivirus expression vector protects against sepsis-induced renal and hepatic dysfunction. Sepsis-triggered inflammatory response and the release of HMGB1 level could be attenuated by miR-agomir administration. We also found that HMGB1 was a direct target of miR-181-5p, and that the overexpression of miR-81-5p led to a significant decrease in HMGB1 protein expression. CONCLUSIONS: miR-181-5p-mediated protective effects in septic mice were modulated, at least partially, through post-transcriptional repression of HMGB1 protein expression. The findings suggest that miR-181-5p may function as an HMGB1 antagonist for alleviating sepsis-induced systemic inflammatory diseases.
Subject(s)
Disease Models, Animal , HMGB1 Protein/metabolism , MicroRNAs/metabolism , Sepsis/metabolism , Animals , Cells, Cultured , Male , Mice , Mice, Inbred C57BL , Sepsis/pathologyABSTRACT
From July 2018 to April 2019, 10 patients (8 males and 2 females, aged 5 to 53 years) who need skin grafting with 17 hands were admitted to the Fourth Medical Center of PLA General Hospital. The method of glove bandaging was used for wound hemostasis in skin grafting. After the initial hemostasis, sterile latex gloves were used as the inner cover of hand, and then pressure-wrapped with gauze. After 20 minutes, the gauze and gloves were removed in order from the near to the far, then thorough hemostasis was conducted again, and finally skin grafting was performed. The use of gloves as inner cover can avoid the adhesion of gauze fibers to the wound surface, so that it will not cause small blood vessels to re-bleed when the gauze is removed, therefore reducing the excessive use of electrocoagulation and secondary damage to the wound. As a result, blood loss can be reduced and hemostatic time can be shortened.
Subject(s)
Hand , Skin Transplantation , Adolescent , Adult , Bandages , Child , Child, Preschool , Female , Hemostasis , Humans , Male , Middle Aged , Young AdultABSTRACT
ABSTRACT: Bite marks are increasingly common in violent cases such as child abuse and sex crimes. Bite marks are often the result of a suspect's attack or a victim's self-defense. Because human teeth vary in size, shape and arrangement, bite marks on objects such as skin and food are characteristic. By using this principle, forensic odontology can identify or exclude suspects by comparing actual bite marks with the teeth marks in the mouth. In this paper, the practical application of bite mark evidence, the research status and problems of bite mark analysis are briefly reviewed, and the prospect of bite mark analysis is also discussed.
Subject(s)
Bites, Human , Forensic Dentistry , Tooth , Child , Child Abuse , Face , HumansABSTRACT
1. Poultry meat quality is affected by many factors, among which intramuscular fat (IMF) is predominant. IMF content affects tenderness, juiciness and flavour of meat. Krüppel-like transcriptional factors (KLFs) are important regulators of adipocyte differentiation. However, little is known about the KLF9 gene associated with poultry IMF deposition, especially intramuscular adipocyte differentiation.2. Previous work has shown that chicken KLF9 was differentially expressed during adipogenesis of intramuscular preadipocytes differentiation. In this study, the function of KLF9 in chicken intramuscular preadipocytes differentiation was investigated.3. In the chicken preadipocyte differentiation model, KLF9 expression showed a major increase with adipogenic induction. Overexpression of KLF9 down-regulated the expression of the adipogenic marker gene AP2, and impaired triglyceride accumulation. Knockdown of KLF9 in chicken intramuscular preadipocytes increased the expression of PPARG, CEBPA and AP2. In addition, it was proposed that KLF9 may regulate adipogenesis via lncRNAs NONGGAT002209.2, NONGGAT003346.2, NONGGAT000436.2 and NONGGAT006302.2 in chicken.4. The data supported a novel role of KLF9 in regulating chicken intramuscular preadipocyte differentiation. Such findings may contribute to a more thorough understanding of chicken IMF deposition and the improvement of poultry meat quality.
Subject(s)
Adipocytes/cytology , Chickens/physiology , Kruppel-Like Transcription Factors/physiology , Adipocytes/metabolism , Adipose Tissue/cytology , Amino Acid Sequence , Analysis of Variance , Animals , Azo Compounds , Base Sequence , Cell Differentiation , Cells, Cultured , Coloring Agents , Kruppel-Like Transcription Factors/chemistry , Kruppel-Like Transcription Factors/classification , Kruppel-Like Transcription Factors/pharmacology , Meat/standards , Pectoralis Muscles/cytology , Pectoralis Muscles/growth & development , Pectoralis Muscles/metabolism , Phylogeny , Plasmids/genetics , RNA, Messenger/chemistry , RNA, Messenger/isolation & purification , Real-Time Polymerase Chain Reaction/veterinary , Sequence Alignment/veterinary , Staining and Labeling/veterinary , Transfection/veterinaryABSTRACT
Objective: To observe the clinical effects of stage-â ¡ Meek skin grafting on adipose tissue after tangential excision in patients with extensive deep burns, and to explore the functional mechanism. Methods: The medical records of 26 extensively burned patients who met the inclusion criteria and were admitted to the Department of Burns and Plastic Surgery of the Fourth Medical Center of PLA General Hospital from May 2015 to December 2017 were retrospectively analyzed. According to the treatment methods, 14 patients were enrolled in stage-â skin grafting group (10 males and 4 females, aged 27 to 75 years), and 12 patients were enrolled in stage-â ¡ skin grafting group (10 males and 2 females, aged 31 to 76 years). Patients in the 2 groups all underwent debridement of tangential excision, and their healthy adipose tissue was preserved. Meek skin grafting was performed just after tangential excision in patients in stage-â skin grafting group. In patients in stage-â ¡ skin grafting group, porcine acellular dermal matrix (ADM) was applied to cover the wound after tangential excision, and 3 days later, it was removed and Meek skin grafting was performed. The times of complement skin grafting and the wound basic healing time of patients in the 2 groups were observed and recorded. In the stage-â ¡ skin grafting group, the adipose tissue of patients were taken from the wound center immediately after tangential excision and immediately after the removal of porcine ADM, for the observation of structure of the fault surface of adipose tissue through hematoxylin and eosin staining and microvessel density (MVD) through immunohistochemical staining. Data were processed with independent sample t test and Fisher's exact probability test. Results: (1) The times of complement skin grafting of patients in stage-â ¡ skin grafting group was (1.83±0.17) times, which was obviously less than (3.36±0.63) times in stage-â skin grafting group (t=2.19, P<0.05). The wound basic healing time of patients in stage-â ¡ skin grafting group was (35.1±2.3) d, which was obviously shorter than (48.8±4.9) d in stage-â skin grafting group (t=2.27, P<0.05). (2) Immediately after tangential excision, the intercellular substance was few between the adipose cells in adipose tissue of patients in stage-â ¡ skin grafting group. Immediately after the removal of porcine ADM, there was regenerated granulation tissue in the intercellular space of adipose cells of adipose tissue of patients in stage-â ¡ skin grafting group. Immediately after tangential excision, the MVD of adipose tissue of patients in stage-â ¡ skin grafting group was 20.2±1.3 under per 400-time field, which was obviously less than 32.2±1.9 under per 400-time field immediately after the removal of porcine ADM (t=-5.38, P<0.01). Conclusions: Meek skin grafting on the adipose tissue in stage-â ¡ surgery after tangential excision could reduce the times of complement skin grafting and shorten wound healing time of patients with extensive deep burns. The mechanism may be related to the improvement of the recipient condition of adipose tissue.
Subject(s)
Adipose Tissue , Burns/surgery , Skin Transplantation , Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Retrospective Studies , Swine , Wound HealingABSTRACT
OBJECTIVE: The aim of this study was to investigate the expression levels of toll-like receptor 9 (TLR9) and interleukin-23 (IL-23) in renal tissue and serum of patients with lupus nephritis (LN), and to explore their clinical correlation. PATIENTS AND METHODS: LN patients and healthy controls were enrolled in the experimental group and the control group, respectively. Blood samples, serum, and peripheral blood mononuclear cells (PBMCs) were collected. Renal lesion tissues and adjacent normal tissues of LN patients were harvested from a renal tissue biopsy. Serum level of IL-23 was detected by enzyme-linked immunosorbent assay (ELISA). Expression of IL-23 in PBMCs was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Meanwhile, TLR9 expression in renal tissues was accessed by immunohistochemistry staining. Subsequently, 24-h protein urine, renal tubular pathological activity index, erythrocyte sedimentation rate (ESR), serum complement C3 level, and blood albumin level of LN patients were recorded. Also, the correlation between TLR9 expression and these pathological indexes was measured by correlation analysis. RESULTS: Serum level of IL-23 in LN patients was significantly higher than that of healthy controls. Similarly, the mRNA and protein expressions of IL-23 in PBMCs of LN patients were markedly higher than those of healthy controls. IL-23 expression was positively correlated with renal tubular pathological activity index of LN patients. Meanwhile, TLR9 was highly expressed in renal tissues of LN patients. Furthermore, TLR9 expression was positively correlated with 24-h protein urine, renal tubular pathological activity index and ESR, whereas negatively correlated with serum complement C3 level and blood albumin level of LN patients. CONCLUSIONS: IL-23 is highly expressed in the serum of LN patients, and its expression is closely related to the occurrence of LN. Also, the expression of TLR9 is up-regulated in the tubulointerstitium of LN patients, which is correlated with relevant clinical indexes.
Subject(s)
Interleukin-23 Subunit p19/biosynthesis , Lupus Nephritis/metabolism , Toll-Like Receptor 9/biosynthesis , Gene Expression , Humans , Interleukin-23 Subunit p19/genetics , Kidney/metabolism , Kidney/pathology , Lupus Nephritis/genetics , Lupus Nephritis/pathology , Toll-Like Receptor 9/geneticsABSTRACT
Objective:To explore the new mechanism of spectomycin B1 in inhibiting angiogenesis of nasopharyngeal carcinoma and to provide a theoretical basis for targeted gene therapy of nasopharyngeal carcinoma. Method:Human nasopharyngeal carcinoma CNE1 cells were divided into two groups, the control group and spectomycin B1 group. Western blot was used to detect the expression levels of small ubiquitin-related modified proteinï¼SUMOï¼ 1 and vascular endothelial growth factor receptor 2ï¼VEGFR2ï¼. The angiogenesis assay was used to detect the angiogenic ability of CNE1 cells, and the apoptosis was detected by flow cytometry. The model of nasopharyngeal carcinoma-bearing mice was established, spectomycin B1 was administered, tumor volume and weight were measured, and protein expression of CD31 was detected by immunohistochemistry and microvessel density was compared. Result:Spectomycin B1 could reduce deSUMOylation of VEGFR2 protein by 4.05 times, significantly reduce the angiogenic ability of CNE1 cells, and increase the apoptosis rate by 20.68%. In the tumor-bearing mouse model, spectomycin B1 treatment could inhibit subcutaneous tumor growth rate and weight, and the blood vessel density decreased by 40.04%. Conclusion:Spectomycin B1 can inhibit neovascularization of nasopharyngeal carcinoma by inducing deSUMOylation of VEGFR2 protein.
Subject(s)
Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms , Spectinomycin , Vascular Endothelial Growth Factor A/metabolism , Animals , Apoptosis , Mice , Neovascularization, PathologicABSTRACT
Objective:To investigate the efficacy of radical sinus surgery (RSS) on difficult-to-treat rhinosinusitis(DTRS) with nasal polyps. Method: We retrospectively analyzed the clinical data of 26 patients with DTRS that treated with RSS and patients who were not treated with RSS in our hospital from August 2013 to June 2017.The effect of RSS were evaluated by comparing the CT Lund-Kennery score,endoscope Lund-Kennery score,global VAS score and dysosmia VAS score before operation and 6 months after operation. Result: RSS group had more severe Lund-Kennery score,global VAS score and dysosmia VAS score than non-RSS group,and also had more previous surgeries,higher ratio of olfactory region polyps,and higher ratio of co-existing asthma and allergic rhinitis. However RSS group had a lower ratio with pus anot than non-RSS group. The eosinophil count in periheral blood between two groups had no statistical significance. Six months after RSS,the score of endoscope Lund-Kennery,global VAS and dysosmia VAS dependence. 14 patients were successfully cured(53.8%), 12 patients showed improvement(46.2%), no invalid cases. Conclusion: The global symptoms and olfaction of DTRS patients can be improved by RSS combine individual perioperative drug therapy.
Subject(s)
Nasal Polyps/surgery , Rhinitis/surgery , Sinusitis/surgery , Chronic Disease , Endoscopy , Humans , Retrospective StudiesABSTRACT
Objective:To systemically evaluate the clinical significance of continuous transfixion suture for nasal septum in septoplasty. Method:Forty patients with nasal septum deviation were randomly assigned to the suture group including 20 patients who underwent endoscopic septoplasty followed by continuous transfixion suture for nasal septum, or the nasal packing group including 20 patients who underwent endoscopic septoplasty followed by nasal packing. Two groups were compared for the visual analogue scale (VAS) scores for postoperative rhinalgia, headache, lacrimation, dysphagia and sleep disorder, changes in mucociliary transport time (MTT) before and after surgery, and postoperative capillary hemorrhage. Patients were followed up for 2 weeks to observe the short-term postoperative complications. Result:Mean VAS scores for rhinalgia, headache, lacrimation, dysphagia and sleep disorder were all higher in nasal packing group than those in suture group (P<0.05); compared to suture group, there was greater prolongation of MTT before and after surgery in packing group (P<0.05); there was significant difference between two groups in postoperative capillary hemorrhage volume (P<0.05); in the nasal packing group, nasal synechia, nasal dryness and hyposmia were observed in 1, 3 and 2 patients, respectively, within 2 weeks postoperatively, whereas no short-term complications were observed in the suture group. Conclusion:Use of continuous transfixion suture in place of nasal packing following septoplasty can significantly improve the postoperative symptoms, protect nasal mucociliary clearance, and reduce short-term postoperative complications.
Subject(s)
Nose Deformities, Acquired/surgery , Rhinoplasty/methods , Suture Techniques , Humans , Nasal Septum , SuturesABSTRACT
This work studies the expression differences of YKL-40 and TLR4 in nasal sinus mucosa of chronic sinusitis patients with and without nasal polyps and its clinical significances. Fifty chronic sinusitis patients with nasal polyps and 50 chronic sinusitis patients without, accepted by our hospital during February 2016-February 2017, were included and taken as group A and group B, respectively. In addition, another 50 patients with nasal deviation were taken as group C (control group). The ostiomeatal complex mucosa of group A and B and the inferior turbinate mucosa of group C were taken and the fluorescence quantitative PCR method was applied to detect the expression of YKL-40, TLR4 and NF- κB of the mucosa and explore and influence of YKL-40 and TLR4 on NF-κB. There was a negative correlation between YKL-40 and TLR4 in group A, and the difference was statistically significant (P less than 0.05) while there was no relationship between YKL-40 and TLR4 expression in group B. The level of YKL-40 protein in group A was higher than that in group B, which was statistically significant (P less than 0.05). YKL-40 and TLR4 were positively correlated in group A while there was no correlation between YKL- 40 and TLR4 expression in group B. The expression of YKL-40, TLR4 and NF-κB in chronic sinusitis patients with nasal polyps was high. In addition, there was a negative correlation between YKL-40 and TLR4 expression in chronic sinusitis patients with nasal polyps. YKL-40 and TLR4 interacted with each other to activate NF-κB and promote disease progression.
Subject(s)
Chitinase-3-Like Protein 1/biosynthesis , Gene Expression Regulation , Nasal Mucosa/metabolism , Nasal Polyps/metabolism , Sinusitis/metabolism , Toll-Like Receptor 4/biosynthesis , Adult , Aged , Chronic Disease , Female , Humans , Male , Middle Aged , Nasal Mucosa/pathology , Nasal Polyps/pathology , Sinusitis/pathologyABSTRACT
Men with type 2 diabetes (T2D) and obesity are often characterised by low testosterone (T). We aimed to determine whether exenatide (EXE) combined metformin (MET) treatment has a better effect on serum total testosterone (TT) levels than glimepiride (GLI) combined MET treatment in men with T2D and obesity. In a multicentre, 12-week observational study, 176 obese T2D men with failed glycaemic control were included in the study: ninety men (mean age, 43.00 ± 8.50 years) in EXE + MET group and 86 men (mean age, 44.00 ± 7.00 years) in GLI + MET group. Serum TT levels were more significantly increased in EXE + MET group than GLI + MET group (121.72 ± 56.73 ng/dl versus 34.67 ± 16.30 ng/dl). The increasement of TT levels in those patients who lost body weight ≥5% was significantly greater than those who lost weight <5% in the two groups. The changes in TT levels are closely related to the changes in waist circumference (r = -.443, p < .001). Sexual function assessment of EXE + MET group was more significantly improved than GLI + MET group (p < .001). No serious adverse events were observed. In conclusion, short-term combined treatment with EXE and MET is superior to GLI combined MET treatment in the improvement of serum TT levels, which could lead to an improvement of sexual hypofunction in patients with obesity and T2D.
Subject(s)
Anti-Obesity Agents/therapeutic use , Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/therapeutic use , Obesity/drug therapy , Sexual Dysfunction, Physiological/drug therapy , Testosterone/blood , Adult , Anti-Obesity Agents/pharmacology , Blood Glucose/drug effects , Body Weight/drug effects , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/complications , Drug Therapy, Combination/methods , Exenatide/pharmacology , Exenatide/therapeutic use , Humans , Hypoglycemic Agents/pharmacology , Male , Metformin/pharmacology , Metformin/therapeutic use , Middle Aged , Obesity/blood , Obesity/complications , Sexual Dysfunction, Physiological/blood , Sexual Dysfunction, Physiological/etiology , Sulfonylurea Compounds/pharmacology , Sulfonylurea Compounds/therapeutic use , Treatment OutcomeABSTRACT
Objective: To reduce the stemness maintenance ability of endometrial cancer stem cell and increase its sensitivity to chemotherapy by inducing hypoxia inducible factor 1α (HIF-1α) protein deSUMOylation. Methods: Lentiviral plasmid mediated ubiquitin carrier protein 9 (Ubc9) gene silencing was transgened into KLE endometrial carcinoma cells. The expression of Ubc9, small ubiquitin-related modifier 1(SUMO1) and HIF-1α protein was detected by Western blotting. Then tumor stem cells clones were cultured in 96 well plates, and these clone balls diameter were calculated. Cell cycles were determined by flow cytometry. MTT cytotoxicity assay and flow cytometry method were used to test sensitivity of cisplatin to endometrial cancer stem cell. Results: The results of Western blotting showed that Ubc9 gene was silenced well, and the covalent binding state of SUMO-1 and HIF-1α protein levels were significantly decreased (P<0.05). Ubc9 gene silencing in endometrial cancer cells reduced clone formation rate by (31.61±5.29)% down to (11.42±3.07)%, while the cell cycle shift from G1 to G2. IC50 of cisplatin decreased from 44.37 mg/L to 7.39 mg/L, and the rate of cell apoptosis by (41.59±5.37)% down to (26.22±4.03)%. Conclusion: The stemness maintenance ability of endometrial cancer stem cell can be reduced through deSUMOylation of HIF-1α protein by silencing Ubc9 gene expression, and their sensitivity to chemotherapy be enhanced, which provides a new reference for future gene therapy of endometrial carcinoma.
Subject(s)
Cell Hypoxia , Endometrial Neoplasms/pathology , Gene Silencing , Stem Cells/physiology , Antineoplastic Agents/pharmacology , Apoptosis , Cell Line, Tumor , Female , Genetic Therapy , Humans , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Ubiquitin-Conjugating Enzymes/metabolismABSTRACT
Objective: To investigate the effect of AKT1 deSUMOylation induced by Ubc9 silencing on the proliferation and metastasis of hepatocellular carcinoma (HCC) cells. Methods: The Ubc9 gene was silenced using RNA interference, and the expression levels of Ubc9, SUMO1 and AKT1 protein were detected by Western blot. Cell proliferation and cell cycle was analyzed by MTT and flow cytometry. Wound healing and transwell assays were used to detect the cell migration ability. Furthermore, the xenograft model was established, and tumor growth curves were drawn. The in situ apoptotic rates was measured using TUNEL Apoptosis Assay. The expression of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP)-2 and MMP-9 were evaluated by immunohistochemical staining. Results: Knockdown of Ubc9 gene significantly decreased the protein expression levels of Ubc9, conjugated SUMO1, free SUMO1 and AKT1 in HCC cells (P<0.05 for all). In control, siR-neg and siR-Ubc9 groups, the cell proliferation indexes were 53.19%, 54.25% and 39.17%, respectively. Moreover, cell migration distance and migrating cells per low power field for all these three groups were (59.47±4.66) µm and 89.44±8.36, (56.56±5.37) µm and 93.84±8.79, as well as (34.57±6.61) µm and 41.67±5.39, respectively. In the xenograft model, the weights of subcutaneous tumors for these three groups were (3.78±0.69) g, (3.72±0.72) g and (2.09±0.61) g, respectively. The corresponding apoptotic cell rates were (7.79±2.21)%, (6.45±2.48)% and (33.59±5.44)%, respectively. The expression levels of PCNA, MMP-2 and MMP-9 protein were significantly decreased in siR-Ubc9 group (P<0.05). Conclusions: Ubc9 silencing in HCC cells induces AKT1 deSUMOylation, and then inhibits the proliferation and metastasis. These results provide a new therapeutic strategy for liver cancer in the future.
Subject(s)
Carcinoma, Hepatocellular/secondary , Liver Neoplasms/pathology , Proto-Oncogene Proteins c-akt/metabolism , RNA Interference , SUMO-1 Protein/metabolism , Ubiquitin-Conjugating Enzymes/genetics , Animals , Apoptosis , Carcinoma, Hepatocellular/metabolism , Cell Movement , Cell Proliferation , Heterografts , Humans , Liver Neoplasms/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Proliferating Cell Nuclear Antigen/metabolism , Wound HealingABSTRACT
We used next-generation sequencing technology to characterize 19 genomic simple sequence repeat (SSR) markers and 11 expressed sequence tag (EST) SSR markers from Leuciscus leuciscus baicalensis, a small freshwater fish that is widely distributed in Xinjiang, China. Primers were used to test for polymorphisms in three L. leuciscus baicalensis populations in Xinjiang. There were 4-27 (average 11.3) alleles (NA), the expected heterozygosity (HE) was 0.36-0.94 (average 0.75 ± 0.14), the observed heterozygosity (HO) was 0.37-1.00 (average 0.68 ± 0.18), and the polymorphism information content (PIC) was 0.31-0.93 (average 0.71). The averages of HE and PIC for the EST-SSR markers were slightly lower than for the genomic SSR markers. Genetic analysis of the three populations showed similar results for PIC, HE, and NA. Amplifications were performed in nine other species; the top three transferability values were for Rutilus lacustris (80%), Leuciscus idus (76.7%), and Phoxinus ujmonensis (63.3%), with the following average values: PIC (0.56, 4.46, and 0.52); NA (0.40, 3.00, and 0.32); and HO (0.44, 2.74, and 0.22), respectively. L. leuciscus baicalensis is one of the most important commercial fish in Xinjiang, but in recent years, fishery resources have decreased sharply owing to water conservation projects, unreasonable utilization, and invasion by alien species. These novel SSR markers are appropriate for studies involving fingerprinting, gene flow, genetic diversity, population structure, and molecular-assisted breeding, and could contribute to the conservation of L. leuciscus baicalensis.
Subject(s)
Cyprinidae/genetics , Gene Transfer, Horizontal , Microsatellite Repeats , Polymorphism, Genetic , Alleles , Animals , Expressed Sequence Tags , HeterozygoteABSTRACT
Tumor vessels are known to be abnormal, with typically aberrant, leaky and disordered vessels. Here, we investigated whether polarized macrophage phenotypes are involved in tumor abnormal angiogenesis and what is its mechanism. We found that there was no difference in chemotaxis of polarized M1 and M2 macrophages to lewis lung carcinoma (LLC) cells and that either M1 or M2 macrophage-conditioned media had no effect on LLC cell proliferation. Unexpectedly, the M2 but not M1 macrophage-conditioned media promoted the proliferation of human umbilical vein endothelial cells (HUVECs) and simultaneously increased endothelial cell permeability in vitro and angiogenic index in the chick embryo chorioallantoic membrane (CAM). The treatment with M2 but not M1 macrophage-conditioned media increased autophagosomes as well as microtubule-associated protein light chain 3B (LC3-B) expression (a robust marker of autophagosomes) but decreased p62 protein expression (a selective autophagy substrate) in HUVECs, the treatment with chloroquine that blocked autophagy abrogated the abnormal angiogenic efficacy of M2 macrophage-conditioned media. These results were confirmed in urethane-induced lung carcinogenic progression. Urethane-induced lung carcinogenesis led to more M2 macrophage phenotype and increased abnormal angiogenesis concomitant with the upregulation of LC3-B and the downregulation of p62. Clodronate liposome-induced macrophage depletion, chloroquine-induced autophagic prevention or salvianolic acid B-induced vascular protection decreased abnormal angiogenesis and lung carcinogenesis. In addition, we found that the tendency of age-related M2 macrophage polarization also promoted vascular permeability and carcinogenesis in urethane carcinogenic progression. These findings indicate that the M2 macrophages induce autophagic vascular disorder to promote lung cancer progression, and the autophagy improvement represents an efficacious strategy for abnormal angiogenesis and cancer prevention.
Subject(s)
Adenocarcinoma/immunology , Apoptosis/immunology , Carcinogenesis/immunology , Lung Neoplasms/immunology , Macrophages/immunology , Neovascularization, Pathologic/immunology , Adenocarcinoma/chemically induced , Adenocarcinoma of Lung , Animals , Autophagy/immunology , Benzofurans/pharmacology , Capillary Permeability/physiology , Carcinogenesis/chemically induced , Carcinoma, Lewis Lung , Cell Differentiation , Cell Line, Tumor , Cell Movement , Cell Proliferation , Chick Embryo , Chloroquine/pharmacology , Clodronic Acid/pharmacology , Culture Media, Conditioned/pharmacology , Endothelial Cells , Female , Human Umbilical Vein Endothelial Cells , Humans , Lung Neoplasms/chemically induced , Mice , Mice, Inbred C57BL , Microtubule-Associated Proteins/biosynthesis , Phosphoproteins/biosynthesis , Trans-Activators/biosynthesis , Urethane/pharmacologyABSTRACT
We investigated the effect of age on the expression of immune molecules [ANA, C4, double stranded DNA (dsDNA), CD16/32, CD19, CD3, and CD64], urine protein, and pathology in mice with chronic graft-versus-host disease (cGVHD) lupus nephritis (LN), and their relationship with reactivity index score. Mouse models of cGVHD LN were established, and mice were randomly divided into four aged-based groups of nine mice each. Serum levels of ANA, C4, and dsDNA were determined, the urine protein levels were assessed, and expression levels of CD16/32, CD19, CD3, and CD64 were measured. Expression levels of CD16/32+CD19(T1), CD16/CD32+CD3(T2), and CD64+CD3 or CD19(T3) were defined in the thymus, in bone marrow they were defined as CD16/32+CD19(B1), CD16/32+CD3(B2), CD64+CD3 or CD19(B3), and in spleen they were defined as CD16/32+CD19(P1), CD16/32+CD3(P2), CD64+CD3 or CD19(P3), respectively. There were significant differences in the levels of dsDNA and urine protein among the four groups (P < 0.05), which were negatively correlated with age. B1, B2, S1, and S2 were significantly different among the four groups (P < 0.05), with a positive correlation with age for B1 and B2. There was no correlation of expression of ANA, C4, dsDNA, T1-T3, B1-B3, S2-S3 with reactivity index score; S1 was the exception (r = -0.440, P = 0.011). Age influenced levels of dsDNA and urine protein in the mouse cGVHD model of LN. S1 was associated with reactivity index score and might also affect pathological changes.