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1.
Journal of Southern Medical University ; (12): 929-936, 2022.
Article in Chinese | WPRIM | ID: wpr-941023

ABSTRACT

OBJECTIVE@#To investigate effects of physiological hypoxic conditions on suspension and adherence of embryoid bodies (EBs) during differentiation of human induced pluripotent stem cells (hiPSCs) and explore the underlying mechanisms.@*METHODS@#EBs in suspension culture were divided into normoxic (21% O2) and hypoxic (5% O2) groups, and those in adherent culture were divided into normoxic, hypoxic and hypoxia + HIF-1α inhibitor (echinomycin) groups. After characterization of the pluripotency with immunofluorescence assay, the hiPSCs were digested and suspended under normoxic and hypoxic conditions for 5 days, and the formation and morphological changes of the EBs were observed microscopically; the expressions of the markers genes of the 3 germ layers in the EBs were detected. The EBs were then inoculated into petri dishes for further culture in normoxic and hypoxic conditions for another 2 days, after which the adhesion and peripheral expansion rate of the adherent EBs were observed; the changes in the expressions of HIF-1α, β-catenin and VEGFA were detected in response to hypoxic culture and echinomycin treatment.@*RESULTS@#The EBs cultured in normoxic and hypoxic conditions were all capable of differentiation into the 3 germ layers. The EBs cultured in hypoxic conditions showed reduced apoptotic debris around them with earlier appearance of cystic EBs and more uniform sizes as compared with those in normoxic culture. Hypoxic culture induced more adherent EBs than normoxic culture (P < 0.05) with also a greater outgrowth rate of the adherent EBs (P < 0.05). The EBs in hypoxic culture showed significantly up-regulated mRNA expressions of β-catenin and VEGFA (P < 0.05) and protein expressions of HIF-1 α, β-catenin and VEGFA (P < 0.05), and their protein expresisons levels were significantly lowered after treatment with echinomycin (P < 0.05).@*CONCLUSION@#Hypoxia can promote the formation and maturation of suspended EBs and enhance their adherence and post-adherent proliferation without affecting their pluripotency for differentiation into all the 3 germ layers. Our results provide preliminary evidence that activation of HIF-1α/β-catenin/VEGFA signaling pathway can enhance the differentiation potential of hiPSCs.


Subject(s)
Humans , Echinomycin/metabolism , Embryoid Bodies/metabolism , Hypoxia/metabolism , Induced Pluripotent Stem Cells/metabolism , beta Catenin/metabolism
2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 11-17, 2022.
Article in Chinese | WPRIM | ID: wpr-940414

ABSTRACT

ObjectiveTo investigate the effect of Huangqisan pellets (HQS) on the phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway and autophagy in the kidney of diabetic nephropathy (DN) rats. MethodDN rat model was established through high-fat diet combined with intraperitoneal injection of streptozotocin (35 mg·kg-1). DN rats were randomly assigned into model group, irbesartan (0.027 g·kg-1) group, low-dose HQS (0.54 g·kg-1) group and high-dose HQS (1.08 g·kg-1) group. The levels of 24 h urinary total protein (UTP), serum albumin (Alb), serum creatinine (SCr), urea nitrogen (BUN), triglyceride (TG) and total cholesterol (TC) were measured after 12 weeks of continuous administration. The pathological changes of renal tissue were observed via hematoxylin-eosin (HE) staining. The expression of podocyte split diaphragm proteins nephrin and podocin in the renal tissue were detected by immunohistochemistry. The protein levels and phosphorylation of key proteins in PI3K/Akt/mTOR signaling pathway, as well as the expression of yeast Atg6 homolog (Beclin1) and microtubule-associated protein 1 light chain 3 (LC3) in the renal tissue were analyzed by Western blot. ResultCompared with the control group, the model group showcased increased 24 h UTP, SCr, BUN, TG, and TC levels and decreased Alb level (P<0.01). After modeling, the rats showed granulosity of epithelial cells of renal tubules, thickening of capillary basement membrane, proliferation of mesangial cells, and sclerosis of glomerulus. Furthermore, modeling down-regulated the expression of nephrin and podocin in the podocyte hiatus of glomerulus (P<0.01) as well as the protein levels of p-PI3K, p-Akt, and p-mTOR and the autophagy markers LC3 and Beclin1 in renal tissue (P<0.01). Compared with model group, irbesartan and HQS decreased the 24 h UTP, Cr, BUN, TG, and TC levels, increased the Alb level, and alleviated the pathological damage of kidney. Moreover, they up-regulated the expression of Nephrin and Podocin in the podocyte hiatus of glomerulus, as well as the protein levels of p-PI3K, p-Akt, p-mTOR, LC3, and Beclin1 in renal tissue (P<0.05, P<0.01). ConclusionHQS may inhibit the PI3K/Akt/mTOR signaling pathway to enhance podocyte autophagy and protect the glomerulus, thus slowing down the development of DN.

3.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 528-532, 2021.
Article in Chinese | WPRIM | ID: wpr-883776

ABSTRACT

Objective:To investigate the effects of ultrasound-guided intervention with methotrexate combined with lauromacrogol on uterine wall thickness and menstrual recovery time in patients with uterine scar pregnancy.Methods:Sixty patients with uterine scar pregnancy who received treatment in Jinhua Wenrong Hospital, Jinhua Central Hospital, and Yiwu Maternity and Children Hospital from February 2019 to February 2020 were included in this study. They were randomly assigned to receive either uterine artery embolization treatment ( n = 30, control group) or ultrasound-guided interventions with methotrexate combined with lauromacrogol ( n = 30, study group). The success rate of treatment, uterine wall thickness at the scar, menstrual recovery time, hospitalization time, hospitalization expenses, incidence of adverse reactions, serum levels of beta-human chorionic gonadotropin (β-HCG) and high sensitivity C-reactive protein (hs-CRP) after 7 days of treatment and uterine wall thickness before and after treatment were compared between the study and control groups. Results:After treatment, the success rate of treatment in the study group was significantly higher than that in the control group [96.67% (29/30) vs. 70.00% (21/30), χ 2 = 7.680, P < 0.05]. Before treatment, there were no significant differences in β-HCG and hs-CRP levels between the control and study groups ( t = -0.197, 0.197, both P > 0.05). After treatment, serum level of β-HCG in the control and study groups was significantly decreased ( t = 22.803, 10.233, both P < 0.01), and serum level of hs-CRP in the control and study groups was significantly increased ( t = -16.357, 7.854, both P < 0.001) compared with before treatment. After treatment, there were no significant differences in serum β-HCG and hs-CRP levels between the control and study groups ( t = 20.413, -20.306, both P < 0.05). After treatment, the incidence of adverse reactions in the study group was significantly lower than that in the control group [6.67% (2/30) vs. 30.00% (9/30), χ 2 = 5.455, P < 0.05). After treatment, menstrual recovery time, hospitalization time and hospitalization expenses in the study group were shorter or less than those in the control group ( t = 5.080, 5.398, 7.599, all P < 0.05). There was no significant difference in the thickness of uterine wall at the scar site between the two groups ( t = -1.045, P > 0.05). Conclusion:Ultrasound-guided intervention with methotrexate combined with lauromacrogol for treatment of uterine scar pregnancy can effectively increase the success rate of treatment, promote the restoration of menstruation, shorten the length of hospital stay, and is highly safe.

4.
Chinese Journal of Endocrine Surgery ; (6): 67-71, 2019.
Article in Chinese | WPRIM | ID: wpr-743400

ABSTRACT

Objective To investigate the diagnostic value of ultrasound ADNEX model in ovarian tumors.Methods 226 patients with pathological-confirmed ovarian tumor from Jun.2014 to Jun.2018 were selected,and the preoperative ultrasound imaging,clinical and pathological data were analyzed.The ultrasound imaging was recorded,and ovarian tumor was diagnosed through ADNEX model.The diagnostic efficiency of ADNEX model for ovarian benign and malignant tumors was analyzed with pathological findings as the gold standard.Results The differences of onset age,CA125 level,tumor solid components maximum diameter and ratio,>10 locules and ascites were all statistically significant between each ovarian tumor group (P<0.05).The overall accuracy rate of ADNEX model in diagnosing different ovarian tumors was 83.2%,which has a good consistency with histopathological findings (Kappa=0.782,P<0.05).The sensitivity and specificity of the prediction model in predicting benign,borderline,stage Ⅰ,stage Ⅱ-ⅣV and metastatic ovarian tumor were respectively 85.5% and 95.7%,69.2% and 96.0%,81.6% and 93.8%,89.1% and 94.4%,80.0% and 98.5%.Conclusion Ultrasound ADNEX model is helpful in differential diagnosis of benign and malignant ovarian tumor,and it has important significance for clinical treatment and prognosis evaluation.

5.
Chinese Journal of Endocrine Surgery ; (6): 150-153, 2018.
Article in Chinese | WPRIM | ID: wpr-695532

ABSTRACT

Objective To investigate the correlation between the blood flow characteristics of contrastenhanced ultrasound (CEUS) and malignancy of patients with ovarian tumor.Methods 60 cases with ovarian tumors treated by surgery and confirmed by pathology were selected,including 28 cases of ovarian cancer and 32 cases of benign ovarian tumor.The time of arrival (AT),peak time (TTP) and intensity of enhancement (EI) were determined by CEUS,and microvessel density (MVD) of tumor tissue was detected by immunohistochemistry.The correlation of the above parameters with MVD and tumor staging was analyzed.Results AT,TTP of the malignant tumor group (10.8±2.1,25.3±10.5) was significantly lower than those of the benign tumor group (15.4±2.7,59.4±11.2).EI of the malignant tumor group (28.6±2.9) was significantly higher than that of the benign tumor group (14.9±2.6),with statistical significance (P<0.05).MVD of the malignant tumor group (62.8± 13.6) was significantly higher than that of the benign tumor group (21.5±11.4),with significant difference (P<0.05).AT and TTP were negatively correlated with MVD and FIGO staging (r=-0.562,r=-0.504,r=-0.636,r=-0.623),while EI was positively correlated with MVD and FIGO staging (r=0.839 and r=0.785),with statistical difference (P<0.05).Conclusions Blood flow characteristics of CEUS are closely related to the malignancy of ovarian tumors.CEUS can accurately evaluate angiogenesis,blood perfusion and malignancy of ovarian tumors.

6.
Frontiers of Medicine ; (4): 497-508, 2018.
Article in English | WPRIM | ID: wpr-772710

ABSTRACT

Viral infections cause at least 10%-15% of all human carcinomas. Over the last century, the elucidation of viral oncogenic roles in many cancer types has provided fundamental knowledge on carcinogenetic mechanisms and established a basis for the early intervention of virus-related cancers. Meanwhile, rapidly evolving genome-editing techniques targeting viral DNA/RNA have emerged as novel therapeutic strategies for treating virus-related carcinogenesis and have begun showing promising results. This review discusses the recent advances of genome-editing tools for treating tumorigenic viruses and their corresponding cancers, the challenges that must be overcome before clinically applying such genome-editing technologies, and more importantly, the potential solutions to these challenges.


Subject(s)
Humans , Antiviral Agents , Therapeutic Uses , CRISPR-Cas Systems , Carcinoma , Genetics , Therapeutics , Virology , Gene Editing , Genetic Predisposition to Disease , Genetic Therapy , Methods , Tumor Virus Infections
7.
China Journal of Endoscopy ; (12): 6-10, 2018.
Article in Chinese | WPRIM | ID: wpr-702896

ABSTRACT

Objective To explore the application of vaginal ultrasound in laparoscopic myomectomy for the reduction of fibroid residue and reduce the recurrence of fibroids value. Methods 120 patients were randomly divided into observation group and control group according to the patient's admission time, all the patients received laparoscopic surgery, while the observation group operation was cooperated with vaginal ultrasound continued to eliminate the remaining uterine fibroids. After operation, all the patients were followed up for 12 months. The operation of the two groups was observed, the remnants of postoperative myoma and the recurrence of myoma at 12 months after the operation were recorded. Results The observation group's treatment time, blood loss, postoperative exhaust time and hospital stay compared with the control group had no significant difference (P > 0.05). One case of hematoma, one case of ureteral injury, two cases of bladder injury and one case of rectal injury occurred in the observation group. One case of hematoma, one case of bladder injury, two cases incision infection and two cases of rectal injury occurred in the control group. Comparing the incidence of intraoperative complications between the two groups show no significant difference (P > 0.05). The average diameter of fibroids in the observation group was significantly smaller than that in the control group (P < 0.05). In the observation group, only 4 patients showed fibroid remnants that were significantly smaller than those in the control group (P < 0.05). One case of recurrence occurred in the observation group at 9 months and 12 months after operation, while in the control group, at 3, 6, 9, and 12 months after the operation 1, 2, 5, and 7 cases recurred, respectively. The patients in the observation group had a significantly lower incidence of recurrence than those in the control group (P < 0.05). Conclusion The use of colposcopy in patients underwent laparoscopic myomectomy to guide the removal of residual fibroids can significantly reduce the residual fibroids, it is worth to clinically promote.

8.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 896-900, 2017.
Article in Chinese | WPRIM | ID: wpr-611731

ABSTRACT

Objective·To construct C-shaped cartilage rings by rabbit auricular cartilage-derived chondrocytes combing with both electrospun gelatin/ polycaprolactone(GT/PCL) nanofibrous membranes and 3D printed supporters for repairing tracheal cartilage defects.Methods·Primary chondrocytes were isolated from rabbit auricular cartilage with methods of trypsin enzyme digestion and collagenase enzyme digestion.After proliferation in vitro,the chondrocytes of passage 2 were harvested for further experiments.Ultrafine composite fibers of GT/PCL were fabricated via electrospinning.The electrospun GT/PCL membranes were tailored into rectangle shape,the length of which is 12 cm and the width is 2.5 cm.Chondrocytes were seeded on membrane at a density of 1 × 108 cells/mL.Then the membrane were rolled onto a 3D printed supporter of poly(DL-lactide-ε-caprolactone) (PLCL) material to construct a C-shaped cartilage-like complex.After 8 weeks of subcutaneous incubation in vivo,gross inspection and paraffin section staining were applied for evaluation.Results·After 8 weeks of culture in vivo,mature cartilage-like tissue were formed with open-cylindrical bellow appearance and pecific mechanical property.C-shaped rings arranged at regular intervals on the inner surface of tissue,which were similar to the normal structure of tracheal cartilages.Histological and immunohistological staining showed a large number of typical lacunar structures and extracellular matrix secretions.Conclusion·It is feasible to construct tissue engineered C-shaped cartilage tissue by combing chondrocytes with GT/PCL membrane and 3D printed PLCL supporter for tracheal cartilage repair.

9.
Chinese Journal of Burns ; (6): 411-415, 2010.
Article in Chinese | WPRIM | ID: wpr-305580

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the mechanisms of pressure intervention, and to explore the most effective regime for pressure therapy.</p><p><b>METHODS</b>Several trials were carried out to study the efficacy and mechanism of pressure therapy, and the development and application efficacy of a smart pressure monitored suit (SPMS) for scar management. (1) Effectiveness of pressure therapy. Forty-five patients suffered burn on extremities were divided into pressure treatment group (n = 36) and control group (n = 9) according to the random number table. Patients in pressure treatment group were prescribed with a regime of wearing custom pressure garment (10% strain rate of pressure + 9 mm thick local pressure padding) more than 23 hours per day, while no active intervention was conducted on patients in control group. Scar conditions were assessed using the Vancouver Scar Scale (VSS), spectrocolorimeter, and tissue palpation ultrasound system. Data were processed with t test or paired t test. (2) Changes in fibroblasts growth rate under pressure. Fibroblasts extracted from scar tissue excised during surgery were loaded with 0, 1.1, 2.8, 5.6 mm Hg (1 mm Hg = 0.133 kPa) pressure respectively to observe the growth rate of fibroblasts. Data were processed with Fisher LSD post-hoc analysis. (3) Scar thickness upon pressure. The changes in scar thickness upon 0, 5, 15, 25, 35 mm Hg pressure were measured at early stage (1 - 6 months), mid-stage (7 - 12 months), and late stage (more than 12 months) using the high frequency ultrasound imaging system. Data were processed with correlation analysis and regression analysis. (4) Study on application of SPMS. Thirty-six patients with hypertrophic scars once treated with the conventional garment were recruited and they were prescribed with the regime of wearing SPMS for one month. Feedback from all participants in rating conventional garment and SPMS was obtained using self-reported questionnaire. The interface pressure of pressure garment was measured using the Pliance X system. Data were processed with Wilcoxon Sign-Ranks test.</p><p><b>RESULTS</b>(1) Scar thickness, color, and VSS score were significantly improved in pressure treatment group after two-month of pressure intervention. VSS score of the scars in pressure treatment group was lower than that in control group two months after treatment. (2) The growth rate of scar fibroblasts under 5.6 mm Hg pressure was obviously lower than that under 0 mm Hg pressure 2 days after pressure loading (mean deviation = 0.086, P = 0.001). Growth rates of fibroblasts under 2.8 and 5.6 mm Hg pressure were obviously lower than that under 0 mm Hg pressure 3 days after pressure loading (with mean deviation respectively 0.060 and 0.118, P = 0.003, P < 0.001). (3) Scar thickness was significantly reduced upon pressure, and a negative relationship between scar thickness and pressure level was observed (r = -0.96, P < 0.01). (4) The results of SPMS study showed a reduction in both static pressure (19.5%) and dynamic pressure (11.9%) after one month of usage; while there was nearly 50.0% reduction in pressure in conventional garment. SPMS was rated significantly higher than conventional garment in terms of comfort, permeability and clinical efficacy (P ≤ 0.001).</p><p><b>CONCLUSIONS</b>Pressure therapy can effectively inhibit the growth of hypertrophic scar, while its exact mechanism needs further study for verification. SPMS is convenient to apply for patients. It takes less time to fabricate and adjust when compared to the conventional garment. Its clinical effect is positive and it may expand its application to other medical conditions.</p>


Subject(s)
Humans , Burns , Cicatrix, Hypertrophic , Therapeutics , Compression Bandages , Pressure
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