Objective:To investigate the effects of vitrification and transplantation on mouse ovarian tissues.Methods:Female ICR mice were divided into three groups: fresh-control group ( in vitro oocyte maturation and fertilization, n=9), fresh-transplanted group (ovarian tissue transplantation, in vitro oocyte maturation and fertilization, n=9), frozen-transplanted group (ovarian tissue vitrification-thawed-transplantation, in vitro oocyte maturation and fertilization, n=9). Additionally, frozen-thawed group (ovarian tissue vitrification-thawed, n=6) and ovariectomy group (ovariectomy, n=6) were also set up, to more directly explain the effects of vitrification and freezing on the number of follicles and the changes of endocrine function after ovarian transplantation. We determined the number of follicles by hematoxylin-eosin (HE) staining, neovascularization by CD31 immunohistochemical staining, tissue fibrosis by Masson staining, and serum sex hormone levels by enzyme-linked immunosorbent assay (ELISA) three weeks following ovarian tissue transplantation. In addition, we counted the number of oocytes obtained, oocytes in vitro fertilized, as well as blastocysts. Results:The number of total follicles in both the fresh-transplanted group and the frozen-transplanted group significantly decreased compared with the fresh-control group (all P<0.001) and the frozen-thawed group (all P<0.001). The CD31-positive rate of ovarian tissues in the fresh-transplanted group was significantly higher than that in the fresh-control group ( P=0.044). Although the CD31-positive rate was higher in the frozen-transplanted group than in the fresh-control group, there was no statistical distinction ( P=0.162). The fibrosis area percentage of ovarian tissues in both the fresh-transplanted group and the frozen-transplanted group increased significantly compared with the fresh-control group ( P=0.004; P=0.005). Serum estradiol level in the fresh-transplanted group and the frozen-transplanted group was significantly lower than that in fresh-control group ( P=0.005; P=0.001), significantly higher than that in the ovariectomy group ( P=0.011; P=0.035). Serum follicle-stimulating hormone (FSH) level in the fresh-transplanted group and the frozen-transplanted group was significantly higher than that in the fresh-control group ( P=0.040; P=0.012), significantly lower than that in the ovariectomy group ( P=0.001; P=0.004). In comparison to the fresh-control group, the number of oocytes retrieved in the fresh-transplanted group and the frozen-transplanted group decreased significantly (all P<0.001). Furthermore, the number of oocytes retrieved in the fresh-transplanted group was higher than that in the frozen-transplanted group, and yet there was no statistical difference ( P=0.272). And the number of oocytes in vitro fertilized and blastocysts in the fresh-transplanted group and the frozen-transplanted group were significantly lower than those in the fresh-control group (all P<0.001). The number of total follicles, CD31-positive rate, fibrosis area rate, serum estradiol, and FSH levels, the number of oocytes in vitro fertilized and blastocysts were no substantially distinction between the fresh-transplanted group and the frozen-transplanted group (all P>0.05). Conclusion:?After ovarian tissue vitrification-thawed and transplantation, follicular growth, endocrine function, and fertility were restored in the mouse model, confirming that ovarian tissue vitrification is an effective method for female fertility preservation. Both vitrification and transplantation could cause follicles to be lost and fertility to decrease. And post-transplantation stage is the primary stage of follicle loss during ovarian tissue frozen-thawed-transplantation, and transplantation is the predominant factor affecting the effectiveness of ovarian tissue frozen-thawed-transplantation.

Objective:To investigate the effects of vitrification and transplantation on mouse ovarian tissues.Methods:Female ICR mice were divided into three groups: fresh-control group ( in vitro oocyte maturation and fertilization, n=9), fresh-transplanted group (ovarian tissue transplantation, in vitro oocyte maturation and fertilization, n=9), frozen-transplanted group (ovarian tissue vitrification-thawed-transplantation, in vitro oocyte maturation and fertilization, n=9). Additionally, frozen-thawed group (ovarian tissue vitrification-thawed, n=6) and ovariectomy group (ovariectomy, n=6) were also set up, to more directly explain the effects of vitrification and freezing on the number of follicles and the changes of endocrine function after ovarian transplantation. We determined the number of follicles by hematoxylin-eosin (HE) staining, neovascularization by CD31 immunohistochemical staining, tissue fibrosis by Masson staining, and serum sex hormone levels by enzyme-linked immunosorbent assay (ELISA) three weeks following ovarian tissue transplantation. In addition, we counted the number of oocytes obtained, oocytes in vitro fertilized, as well as blastocysts. Results:The number of total follicles in both the fresh-transplanted group and the frozen-transplanted group significantly decreased compared with the fresh-control group (all P<0.001) and the frozen-thawed group (all P<0.001). The CD31-positive rate of ovarian tissues in the fresh-transplanted group was significantly higher than that in the fresh-control group ( P=0.044). Although the CD31-positive rate was higher in the frozen-transplanted group than in the fresh-control group, there was no statistical distinction ( P=0.162). The fibrosis area percentage of ovarian tissues in both the fresh-transplanted group and the frozen-transplanted group increased significantly compared with the fresh-control group ( P=0.004; P=0.005). Serum estradiol level in the fresh-transplanted group and the frozen-transplanted group was significantly lower than that in fresh-control group ( P=0.005; P=0.001), significantly higher than that in the ovariectomy group ( P=0.011; P=0.035). Serum follicle-stimulating hormone (FSH) level in the fresh-transplanted group and the frozen-transplanted group was significantly higher than that in the fresh-control group ( P=0.040; P=0.012), significantly lower than that in the ovariectomy group ( P=0.001; P=0.004). In comparison to the fresh-control group, the number of oocytes retrieved in the fresh-transplanted group and the frozen-transplanted group decreased significantly (all P<0.001). Furthermore, the number of oocytes retrieved in the fresh-transplanted group was higher than that in the frozen-transplanted group, and yet there was no statistical difference ( P=0.272). And the number of oocytes in vitro fertilized and blastocysts in the fresh-transplanted group and the frozen-transplanted group were significantly lower than those in the fresh-control group (all P<0.001). The number of total follicles, CD31-positive rate, fibrosis area rate, serum estradiol, and FSH levels, the number of oocytes in vitro fertilized and blastocysts were no substantially distinction between the fresh-transplanted group and the frozen-transplanted group (all P>0.05). Conclusion:?After ovarian tissue vitrification-thawed and transplantation, follicular growth, endocrine function, and fertility were restored in the mouse model, confirming that ovarian tissue vitrification is an effective method for female fertility preservation. Both vitrification and transplantation could cause follicles to be lost and fertility to decrease. And post-transplantation stage is the primary stage of follicle loss during ovarian tissue frozen-thawed-transplantation, and transplantation is the predominant factor affecting the effectiveness of ovarian tissue frozen-thawed-transplantation.

Two patients (patient 1, a 63-year-old female; patient 2, a 22-year-old male) were treated with anlotinib for lung adenocarcinoma and synovial sarcoma, respectively. Patient 1 developed cough, expectoration, and dyspnea after 8 cycles of anlotinib treatment (12 mg orally once daily, 2 weeks of treatment and 1 week of withdrawal as a treatment cycle). Spontaneous pneumothorax was diagnosed by chest CT. The spontaneous pneumothorax was considered to be related to anlotinib. Then the drug was stopped. After thoracic cavity closed drainage therapy, the patient′s dyspnea was relieved. Due to the progress of the disease, the patient received reduced-dose anlotinib treatment (10 mg/d, with the same usage as before). Spontaneous pneumothorax recurred in the first cycle. Anlotinib treatment was terminated and the symptomatic treatments including thoracic cavity closed drainage, expectorant, anti-infection, and nutritional support were given. Ten days later, chest X-ray examination showed that pneumothorax was cured. Patient 2 developed spontaneous pneumothorax (found by chest CT examination) after 9 cycles of treatment with anlotinib (12 mg/d, with the same usage as that in patient 1). But the patient had no obvious discomfort and continued to use anlotinib under close monitoring of respiratory status.

Two patients (patient 1, a 63-year-old female; patient 2, a 22-year-old male) were treated with anlotinib for lung adenocarcinoma and synovial sarcoma, respectively. Patient 1 developed cough, expectoration, and dyspnea after 8 cycles of anlotinib treatment (12 mg orally once daily, 2 weeks of treatment and 1 week of withdrawal as a treatment cycle). Spontaneous pneumothorax was diagnosed by chest CT. The spontaneous pneumothorax was considered to be related to anlotinib. Then the drug was stopped. After thoracic cavity closed drainage therapy, the patient′s dyspnea was relieved. Due to the progress of the disease, the patient received reduced-dose anlotinib treatment (10 mg/d, with the same usage as before). Spontaneous pneumothorax recurred in the first cycle. Anlotinib treatment was terminated and the symptomatic treatments including thoracic cavity closed drainage, expectorant, anti-infection, and nutritional support were given. Ten days later, chest X-ray examination showed that pneumothorax was cured. Patient 2 developed spontaneous pneumothorax (found by chest CT examination) after 9 cycles of treatment with anlotinib (12 mg/d, with the same usage as that in patient 1). But the patient had no obvious discomfort and continued to use anlotinib under close monitoring of respiratory status.

Objective To study the impact of interposed abdominal pulling-pressing cardiopulmonary resuscitation (IAPP-CPR) for patients with cardiac arrest (CA). Methods A prospective study was conducted. A total of 122 CA patients admitted to Department of Emergency of Shandong Provincial Mining Industry Group Company Central Hospital from July 2013 to December 2017 were enrolled. They were divided into standard cardiopulmonary resuscitation (S-CPR) group (n = 62) and IAPP-CPR group (n = 60) according to order of admission. The patients in S-CPR group received external cardiac compression, open airway, endotracheal intubation, mechanical ventilation, routine drug rescue, and defibrillation when ventricular fibrillation was found. And the patients in IAPP-CPR group received the IAPP-CPR on the basis of the routine chest compression. During the relaxation period, the patients were subjected to abdominal lifting and compressing with amplitude of 4-5 cm, frequency of 100 times/min, and the time ratio of lifting to compressing was 1:1. The data of demographics and clinical signs of patients were collected. The markers of respiratory and circulatory performance of all patients after CPR were determined. The rates of restoration of spontaneous circulation (ROSC), successful resuscitation, and the prognosis were recorded. With the success of CRP as the dependent variable, the factors with statistical significance showed by univariate analysis were used as the independent variable to carry out two classification Logistic regression analysis for screening the influence factors of CPR success. Receiver operating characteristic (ROC) curve was plotted to analyze the predictive value of various factors on the success of CPR. Results 122 patients were enrolled in the analysis. Compared with the S-CPR group, heart rate (HR), mean arterial pressure (MAP), arterial partial pressure of oxygen (PaO2), and end-tidal carbon dioxide partial pressure (PETCO2) were significantly increased at 30 minutes after CPR in IAPP-CPR group [HR (bpm): 66.3±11.5 vs. 53.1±12.6, MAP (mmHg, 1 mmHg = 0.133 kPa): 65.4±6.5 vs. 53.2±5.4, PaO2(mmHg): 77.7±11.8 vs. 61.8±14.3, PETCO2(mmHg):45.5±9.6 vs. 31.8±8.2, all P < 0.05], and arterial partial pressure of carbon dioxide (PaCO2) and lactic acid (Lac) were significantly lowered [PaCO2(mmHg): 46.7±6.2 vs. 57.9±9.5, Lac (mmol/L): 2.1±1.5 vs. 4.4±2.2, both P < 0.05]. The time of CA to ROSC in IAPP-CPR group was significantly shorter than that in S-CPR group (minutes: 6.3±1.8 vs. 11.2±1.4, P < 0.05), the ROSC rate and CPR success rate were significantly higher than those in S-CPR group [ROSC rate: 61.7% (37/60) vs. 43.5% (27/62), CPR success rate: 40.0% (24/60) vs. 21.0% (13/62), both P < 0.05], and 24-hour survival rate and survival and discharge rate of patients were significantly higher than those in the S-CPR group [24-hour survival rate: 46.7% (28/60) vs. 29.0% (18/62), survival and discharge rate: 20.0% (12/60) vs. 11.3% (7/62), both P < 0.05]. Logistic regression analysis showed that PaO2, PaCO2 and PETCO2 were the factors that affect the success of CPR [PaO2: β= -3.76, odds ratio (OR) = 0.23, 95% confidence interval (95%CI) = 0.12-0.86, P = 0.031; PaCO2:β=1.41,OR=4.09,95%CI=1.70-9.82,P=0.002,PETCO2:β=0.78,OR=2.18,95%CI=1.42-3.35,P=0.000]. ROC curve analysis showed that the above three factors had good predictive value for the success of CPR. The predictive value of PaCO2 and PETCO2 were better, the area under ROC curve (AUC) was 0.93 and 0.92, respectively, when the cut-off values was 46.7 mmHg and 48.8 mmHg, the sensitivity was 92.0%, 88.0%, respectively, and the specificity was both 94.3%. Conclusions PaO2, PaCO2 and PETCO2 are the factors that influence the success of CPR. PaCO2 and PETCO2 have great value in predicting the success of CPR. Compared with the S-CPR group, IAPP-CPR group results in better hemodynamic and pulmonary ventilation effects, and remarkably improve ROSC and successful resuscitation. IAPP-CPR has obvious clinical value for CA patients.

Objective To study the expression of early growth response gene-1(Egr-1)in esophageal carcinoma tissue,and to explore the relationship between Egr-1 and the survival time of the patients with esophageal carcinoma.Methods RT-PCR was performed to detect the expression of Egr-1 mRNA in68 cases of esophageal carcinoma.The relationship between survival time and prognosis was analyzed.Results The expression of Egr-1 mRNA was the lowest(0.567±0.404),(0.945±0.336)and(1.201±0.347)in esophageal carcinoma tissue,para-cancerous tissue and normal esophageal mucosa tissue,respectively(F=12.709,P＜0.05,P＜0.00).21 cases showed the positive expression of Egr-1 mRNA of both the esophageal carcinoma tissue and the para-cancerous tissue.21 cases showed the positive expression of Egr-1 mRNA in a single esophageal carcinoma tissue or the para-cancerous tissue.26 cases showed the negative expression of Egr-1 mRNA both in the esophageal carcinoma tissue and the paracancerous tissue.The positive rate of Egr-1mRNA expression was 65.71%and 30.00%in the groups of the survival time for three years and the groups of the survival time for one year(P＜0.05).The survival rates in the two groups with positive expression of Egr-1 mRNA were 94.44%and 86.96%,respectively(P＞0.05).Conclusion Decreased level of EGR1 expression may be related to esophageal carcinogenesis.The expression level of Egr-1 mRNA might be associated with the survival time of the patients with esophageal carcinoma.Egr-1 expression in esophageal carcinoma tissue may be of great value for determining prognosis of esophageal carcinoma.

Objective To explore the relationship between the apoptosis of esophageal carcinoma tissue and its adjacent tissues with the survival time of patients with esophageal carcinoma by examining the apoptosis in esophageal carcinoma tissue and its adjacent tissues.Methods FCM Wag performed to detect the rates of apoptosis in 68 cases of esophageal carcinoma and their adjacent tissues and 28 cases of normal esophageal mucosae tissue.Sixty-three patients with esophageal carcinoma had been followed up and noted the survival time of every patient from the operated day to the deadline.Results The rate of apoptosis was the highest in the normal esophageal mucosae tissue(12.78±1.32)%,(7.79±1.48)% in adjacent tissue,and(4.16±2.06)% in esophageal carcinoma tissue,respectively.To follow the survival time after operation of 63 cases with esophageal carcinoma showed 60 cases in one year survival time and 35 cases in three years survival time.The rate of apoptosis in the esophageal carcinoma and its adjacent tissues wag(3.45±1.51)% and (3.96±0.94)% in the patients of one year survival time,(3.90±2.53)% and (7.89±2.27)% in the patients of three years survival time,respectively,P<0.05.Conclusions There is the phenomenon of apoptosis-escape in the esophageal carcinogenesis.There is close relationship between the apoptosis of esophageal carcinoma tissue and the survival time after operation.The apoptosis in the adjacent tissue is more important than that in the esophageal carcinoma tissue for evaluating the survival time after operation of patients with esophageal carcinoma.

Objective: To investigate the effect of s urface modification of stearic acid solid lipid nanoparticles with Brij 78 on the body distribution. Me thods: T he stearic acid solid lipid nanoparticles were prepared by “emulsi on-evaperation-solidfied at low temperature" method with two kinds of surfacta nts: Brij 78 and taurocholic acid sodium salt. Groups of five randomly selected Kun Ming mice of both sexes were used for each suspension and for each time point. After injection of each suspension into the tail vein, one group of mice were killed a t e ach time point. Then the blood, liver, spleen, lung, heart, kidney, muscles and fat were removed and the dpm value of 100 μl blood and 50 mg of each tissue was determined. Results: Compared with the nanoparticles taurocholi c acid sodium salt, the AUC (tissue)/AUC(blood) value of to spleen, lung, heart and kidney with the nanoparticles modified with Brij 78 was increased to 1 12.5%, 143.6%, 146.7% and 184.8% respectively. Conclusion: The distribution of stearic acid solid l ipid nanoparticles into the non-RES organs can be increased and the circulating time may be prolonged by modifying the nanoparticle surface with Brij 78.

Objective To investigate the effects and the gene expression of mitochondria on arsenic trioxide-induced apoptosis of acute promyelocytic leukemia (APL) NB4 cells. Methods NB4 cells were treated with As2O3. Fluorescent microscopy and flow cytometry were used and mitochondria membrane potential detection and RT-PCR were performed to observe the NB4 cell apoptosis, growth inhibitory effect and mitochondrial transmembrane potentials. Mitochondria genome primers were designed, and the expression of mitochondria genome at gene and protein levels was studied. Results Induced by As2O3, the NB4 cells showed typical morphological changes of apoptosis with a significant growth inhibitory effect. The ratios of NB4 cells apoptosis were 5.02%, 6.40%, 28.40% and 33.34%, respectively, when treated with As2O3 in concentrations of 0.5?mol/L, 1?mol/L, 2?mol/L and 3?mol/L. When treated with As2O3 in 0.5?mol/L, 1?mol/L, 2?mol/L, 4?mol/L and 8?mol/L at 48h, the mitochondria potential of NB4 cells was decreased by 12.8%, 21.6%, 66.9%, 83.7% and 83.8%, respectively. After the NB4 cell apoptosis was induced by As2O3, RT-PCR assay was used to detect the expression of 13 genes of mitochondria genome. The expression of COX2 gene was down-regulated in this process, while no change was found in the expression of other 12 genes. Conclusions As2O3 has shown to exert a significant effect on promoting apoptosis and growth inhibition on APL cells. The apoptotic effect induced by As2O3 on NB4 cells is closely related to a decrease of mitochondrial membrane potential. The expression change in the mitochondria gene COX2 is involved in the As2O3-induced apoptosis of NB4 cells.

Objective Gene microarray technology was used to investigate the differential gene expression of apoptosis related genes in NB4 cells induced by arsenic trioxide. Methods The databases of Evntrez and Human IPI were searched with "apoptosis or apoptotic" as the key words, and 1384 apoptosis related genes were found after the redundant genes were eliminated by chromosomal localization. The probes of these genes were designed using OligoArray 2.0, and then analyzed by BLAST. All the probes were immobilized on the glass slide, which were used as oligonucleotide microarray. After NB4 cells were treated with 2umol/L As2O3 for 48h, the total RNA were extracted. cDNAs of control group and test group were fluorescently labeled with Cy3 and Cy5, respectively, by RT-PCR. The fluorescent samples were hybridized with an oligonucleotide microarray containing 1384 apoptosis related genes to search for the differentially expressed genes in the cells with or without As2O3 treatment. The hybridization signals were scanned by oligonucleotide microarray, and then the fluorescent intensity of Cy3 and Cy5 and the ratio of two fluoresceins were analyzed using certain software. Then the differential expressed genes were analyzed after As2O3 treatment, in which the most distinctly differential expressed genes were chosen as targets, and through PCR amplification and gel electrophoresis the above genes were verified. Results There are 4 genes up-regulated and 12 genes down-regulated in expression in NB4 cells after 48h treatment with 2umol/L As2O3, which were in accordance with the results of RT-PCR and oligonucleotide microarray. Conclusion Differential gene expression in NB4 cells was induced by As2O3 treatment. These differentially expressed genes, with relation to signal transduction, transcription regulation, cell cycles, oxidation response, protein translation and cell differentiation, may play an important role in NB4 cell apoptosis.