Objective To prepare the sinomenine hydrochloride multivesicular liposomes with high entrapment efficiency and sustained release character.Methods Multiple emulsion method was used to prepare the sinomenine hydrochloride multivesicular liposomes.Uniform design was applied to optimize the formulation and pharmaceutical process.The shape,the particle size,and the release charcter of the liposome were evaluated.Results The sinomenine hydrochloride multivesicular liposomes prepared were spherical and the size of majority particles was in the range of 20—30 ?m and well distributed.The encapsulation efficiency was more than 80% and its in-vitro release profile accorded well with the Higuchi model with t1/2 up to 52.7 h.Conclusion The formulation and pharmaceutical process of the sinomenine hydrochloride multivesicular liposomes are stable and feasible with the high encapsulation efficiency and good sustained-release character.

Objective To reduce the incidence of the hypocapnia,the cutoff value of the end-tidal carbon dioxide partial pressure[Pet(CO2)] for predicting the hypocapnia so as to understand the suitable adjustment target and target range of the Pet(CO2) in preterm infants under mechanical ventilation.Methods From Jan.2012 to Oct.2013,96 cases of the preterm infants with respiratory distress syndrome(RDS) who needed mechanical support were selected from the Huaian Maternity and Child Health Care Hospital.Pet(CO2) value of each time point(1 h,24 h,48 h and 72 h after mechanical ventilation) were recorded,while radial artery blood was collected for blood gas analysis.The level of pa (CO2) ＜ 35 mmHg(1 mmHg =0.133 kPa) diagnosed hypocapnia;while the level of Pa (CO2) ＞ 60 mmHg was for diagnosing hypercapnia.The diagnostic cutoff and the suitable adjustment target and adjustment target range of the Pet(CO2) were confirmed by receiver operating characteristic (ROC) curve.Results The data from 381 arterial blood gas analysis results were gained,of which 151 times belonged to hypocapnia,and the rate was 39.6％,the other 230 cases were normal,and no case was of hypercapnia.The area under the ROC curve was 0.895,and the area of the standard error was 0.016.There was a statistical significance in Pet(CO2) value for the diagnosis of hypocapnia(P =0.000).The lower the value of Pet (CO2),the greater the likelihood of hypocapnia,and 95 ％ confidence interval area was 0.864-0.926.The Pet (CO2) optimal diagnostic cutoff value determined in accordance with Youden index was 30.5 mmHg.When Pet (CO2) among 41.5 mmHg,sensitivity was 100％.Conclusions Diagnostic cutoff value for forecasting hypocapnia is 30.5 mmHg.The suitable adjustment target of mechanical ventilation parameter adjustment is 41.5 mmHg for the Pet(CO2).The target range of mechanical ventilation parameter adjustment is 30.6-41.5 mmHg for the Pet(CO2).

Objective To prepare sinomenine hydrochloride transfersomes and evaluate their qualities. MethodsThree different preparation methods including film dispersion, reverse phase evaporation, and ethanol injection methods were compared according to the encapsulation efficiency of transfersomes. Uniform design was applied to optimize the formulation and pharmaceutical process of reverse phase evaporation. The particle size, the appearance, the Z-potential, and the stability were also evaluated. ResultsThe transfersomes prepared by reverse-phase evaporation method possessed the highest encapsulation efficiency. The ideal combinations of preparation and formulation were: soya lecithin/sodium cholate was 200/30 mg/mg, chloroform/PBS was 5 mL/mL, pH of PBS was 6.5, added sinomenine hydrochloride was 10 mg. The transfersomes obtained were milky white translucent suspension, with a mean encapsulation efficiency of 62.2%. The shape of their particles was spherical or similar to spherical under microscope, which was smooth and disconglutinated with an average diameter of 96.4 nm, and a Z-potential of-35.93 mV. Aggregation or deposition was not observed after exposure under the temperature of 4 ℃ for 30 d. ConclusionThe preparation process of sinomenine hydrochloride transfersomes is feasible, the quality of obtained transfersomes is stable.It is expected to provide a new preparation for clinical use of sinomenine hydrochloride.

Objective To explore the clinical significance of Downes score in predicting respiratory failure (RF) in term neonates.Methods From June 1, 2011 to December 31, 2013, 243 hospitalized term neonates with dyspnea, who were born at 37 to 42 gestational age and admitted to the hospital within 24 h after birth, were selected from Neonatal Medical Center of Huai'an Maternity and Child Healthcare.Downes scores were evaluated based on five indicators including respiratory rate, skin color, three depressions sign, moaning and lung breath sounds at the time of admission.RF was diagnosed based on clinical menifestations and radial arterial blood gas analysis.Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive value of Downes score on RF.Results The average Downes score of the 243 cases was 6.57±0.86 and the RF rate was 69.6％ (169/243).The RF rate was 93.4％ (169/181) when Downes score ≥ 4.5, with the sensitivity, specificity, misdiagnostic rate and missed diagnositic rate of 100.0％, 83.8％, 16.2％ and 0.0％, respectively.While, the RF rate was 80.7％ (130/161) when Downes score ≥ 5.5, with the sensitivity, specificity, misdiagnostic rate and missed diagnositic rate of 76.9％, 58.1％, 41.9％ and 23.1％, respectively.When Downes score ≥ 6.5, the incidence of RF was 97.6％ (123/126), with the sensitivity, specificity, misdiagnositic rate and missed diagnositic rate of 72.8％, 95.9％, 4.1％ and 27.2％, respectively.The cutoff value was 6.5 according to the Youden Index, which with lower rate of misdiagnosis and missed diagnosis.When Downes score ≥ 7.5, the incidence of RF was 100.0％(22/22), with the sensitivity, specificity, misdiagnositic rate and missed diagnosis rate of 13.1％, 100.0％, 0.0％ and 87.0％, respectively.The area under the ROC curve was 0.906 (95％CI: 0.866-0.955).The higher the Downes score, the greater the possibility of RF (P=0.000).Conclusions Dynamic Downes scores in term neonates with dyspnea could predict the risk of RF.The higher the Downes score, the greater the possibility of RF.

Objective To observe the clinical effects of therapy to neonatal jaundice by Yinzhihuang particles.Methods72 cases from July 2015 to December 2016, were randomly divided into two groups and made all 36 cases in the control group, children in the control group given blue light therapy observation group were treated in the control group on the basis of Yinzhihuang particle therapy, follow-up and record two serum C-reactive protein (CRP), total bilirubin (TBIL), γ-alanine amino acyltransferase (γ-GT), alkaline phosphatase (AKP) and free fatty acid (FFA) levels and incidence of adverse reactions, the use ofstatistical methods for data analysis.Results① observation group after treatment CRP,TBIL, γ-GT values were better than the control group, and the difference was significant (P<0.05).② observation group in the AKP, FFA values after treatment than the control group, and the difference was significant (P<0.05).③ observation group rash during treatment, the incidence of diarrhea were (0.00%, 2.78%), were lower than the control group (11.11%, 16.67%), and the difference was significant (P<0.05).ConclusionYinzhihuang particle treatment of neonatal jaundice effect is good, worthy of further research and application.

To investigate the reliability and feasibility of human papillomavirus (HPV) DNA test in cervical scraping smears with polymerase chain reaction (PCR), 131 cases of cervical scraping specimens were collected, and the positive rates and accuracy of HPV infection were determined in normal subjects and cervical cancer patients. GP5+/GP6+ and E7 primer pairs designed for detecting HPV L1 and HPV type 16 E7 were tested in this study. Our results showed that positive rates of HPV DNA in normal population and cervical cancer patients were 32.99 % and 73.53 % respectively and there was significant difference between them (P＜0. 001). In normal subjects, detection rates of HPV DNA with GP5+/GP6+ and E7 primer pairs were 27.84 % and 16.49 % respectively, with statistically significant difference between them (P＞0.05). However the detection rates in cervical cancer patients were 38.24 % and 67.65 % for the two markers, with a significant difference found between them (P＜0.05). It is concluded that HPV DNA test with PCR for cervical scraping smears was feasible. GP5+/GP6+ primer pairs may be a useful probe to screen HPV infection in normal population, but they are not sensitive enough in cervical cancer patients. It is suggested that high risk type HPV DNA test was very useful in population with high risk of cervical cancer.

The relationship of connexin43 (Cx43) and bystander effect in ovarian tumor cells in herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapy in vitro was explored and the effect of all-trans retinoic acid (RA) on the expression of Cx43 and bystander effect investigated. The Cx43 expression was detected by flowcytometry, Western blot, and immunofluorescence in two ovarian tumor cell lines OVCAR3, CaOV3 before and after RA treatment. Bystander effect was determined by the cells growth inhibitory rate with methyl thiazolyl tetrazolium. Following exposure to ganciclovir, there was much greater bystander killing in OVCAR3 than that in CaOV3 (P<0.05). The expression of Cx43 was detected in OVCAR3 by flowcytometry and Western blot, but it could not be detected in CaOV3. The expression of Cx43 in both cell lines could be induced by RA. Immunofluoresence staining showed that Cx43 protein of OVCAR3 was located on membrane surface, whereas CaOV3 in cytoplasm. RA could not change the location of Cx43 protein in both cell lines. There is relationship between Cx43 expression and HSV-TK/GCV bystander effect. HSV-TK/GCV bystander effect can be enhanced by RA in ovarian cancer.
Antiviral Agents ; pharmacology ; Bystander Effect ; Cell Line, Tumor ; Connexin 43 ; biosynthesis ; genetics ; Female ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; Genetic Therapy ; Humans ; Ovarian Neoplasms ; metabolism ; therapy ; Pregnancy ; Simplexvirus ; genetics ; Thymidine Kinase ; genetics ; Tretinoin ; pharmacology

Objective:To screen the differentially expressed exosomal miRNAs derived from liver cancer stem cells (LCSCs) and its effect on the malignant biological characteristics of liver cancer cells.Methods:miRNA expression profile chip was used to analyze the differentially expressed exosomal miRNA derived from LCSCs. The effects of miRNA on malignant phenotypes of LCSCs were identified. The cells were further treated with doxorubicin at different concentrations (0, 150, 300 μmol/L), and the expression level of miR-196a was detected by quantitative real-time PCR (qRT-PCR). The apoptosis of liver cancer cells cultured by exosomes derived from LCSCs (Exo-NC group) and exosomes derived from miR-196a inhibited LCSCs (Exo-Inhibitor group) and the activity of caspase3/7 under the action of exosomes from LCSCs were detected. Nude mice were randomly divided into Do-PBS group, Do-Exo-Inhibitor group and Do-Exo-NC group using random number table method, with 5 mice in each group, and the effect of miR-196a on nude mice xenograft tumor model with liver cancer cells was analyzed.Results:In this study, exosomes were isolated and purified from CD133 + Huh7 stem cell culture supernatant. miR-7162-3p, miR-1910-5, miR-3613-3p, miR-196a and miR-155-5p were up-regulated, while miR-1246 and miR-3613-5p were down-regulated. miR-7162-3p, miR-196a and miR-155-5p in exosomes had important effects on the self-renewal ability of LCSCs. miR-1910-5p, miR-196a and miR-155-5p had important effects on the invasion ability of liver cancer stem cells, among which miR-196a had the most significant inhibitory effect. Treatment for 24 h, the miR-196a expression level of the 0, 150 and 300 μmol/L doxorubicin was 0.96±0.05, 1.23±0.05 and 2.33±0.03 respectively, with a statistically significant difference ( F=996.90, P<0.001). Treatment for 48 h, the miR-196a expression level of the 0, 150 and 300 μmol/L doxorubicin were 1.02±0.07, 2.35±0.05 and 2.89±0.55 respectively, with a statistically significant difference ( F=303.00, P<0.001). When the concentration of doxorubicin was 0 and 300 μmol/L, the apoptosis rates of the Exo-NC group were 9.37%±0.19% and 11.64%±0.27%, and those of the Exo-Inhibitor group were were 18.80%±1.91% and 22.79%±1.57%, with statistically significant differences ( t=4.41, P=0.048; t=4.96, P=0.038). When doxorubicin was not used, the ratios of caspase3/7 in the Exo-NC group at 24 h and 48 h were 0.94±0.08 and 0.97±0.09, and those in the Exo-Inhibitor group were 1.56±0.01 and 1.58±0.01, with statistically significant differences ( t=11.41, P=0.008; t=6.07, P=0.026). Under 300 μmol/L doxorubicin, the ratios of caspase3/7 in the Exo-NC group at 24 h and 48 h were 0.95±0.07 and 1.36±0.08, and those in the Exo-Inhibitor group were 2.84±0.08 and 3.20±0.14, with statistically significant differences ( t=24.20, P=0.002; t=15.78, P=0.004). The results of xenograft tumor in nude mice showed that the tumor volumes of Do-PBS, Do-Exo-Inhibitor and Do-Exo-NC groups increased successively, which were (1 051.86±89.90) mm 3, (1 310.91±86.66) mm 3 and (2 185.14± 352.34) mm 3 respectively, with a statistically significant difference ( F=30.28, P<0.001). The weights of the transplanted tumors in the 3 groups increased successively, which were (0.36±0.10) g, (0.39±0.12) g and (0.76±0.16) g respectively, with a statistically significant difference ( F=11.81, P=0.002). The expression of miR-196a in tumors was significantly decreased after miR-196a inhibitor transfection. The expression levels of the 3 groups were 1.05±0.16, 0.38±0.08 and 2.17±0.26, with a statistically significant difference ( F=48.93, P<0.001). Conclusion:The exosomal secreted by LCSCs can enhance the resistance of liver cancer cells to doxorubicin by miR-196a.

Objective To investigate the value of a risk assessment model in predicting venous thromboembolism (VTE) in patients with liver failure after artificial liver support therapy. Methods A retrospective analysis was performed for the clinical data of 124 patients with liver failure who received artificial liver support therapy in Affiliated Drum Tower Hospital of Nanjing University Medical School from March 2019 to December 2021, among whom there were 41 patients with VTE (observation group) and 143 patients without VTE (control group). Related clinical data were compared between the two groups, and the Caprini risk assessment model was used for scoring and risk classification of the patients in both groups. The t -test was used for comparison of continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups; the Mann-Whitney U rank sum test was used for comparison of ranked data between two groups. The logistic regression analysis was used to investigate the independent risk factors for VTE in patients with liver failure after artificial liver support therapy. The receiver operating characteristic (ROC) curve was used to investigate the value of Caprini score and the multivariate predictive model used alone or in combination in predicting VTE. Results The observation group had a significantly higher Caprini score than the control group (4.39±1.10 vs 3.12±1.04, t =6.805, P < 0.001). There was a significant difference between the two groups in risk classification based on Caprini scale ( P < 0.05), and the patients with high risk or extremely high risk accounted for a higher proportion among the patients with VTE. The univariate analysis showed that there were significant differences between the two groups in age ( t =6.400, P < 0.001), catheterization method ( χ 2 =14.413, P < 0.001), number of times of artificial liver support therapy ( Z =-4.720, P < 0.001), activity ( Z =-6.282, P < 0.001), infection ( χ 2 =33.071, P < 0.001), D-dimer ( t =8.746, P < 0.001), 28-day mortality rate ( χ 2 =5.524, P =0.022). The multivariate analysis showed that number of times of artificial liver support therapy (X 1 ) (odds ratio [ OR ]=0.251, 95% confidence interval [ CI ]: 0.111-0.566, P =0.001), activity (X 2 ) ( OR =0.122, 95% CI : 0.056-0.264, P < 0.001), D-dimer (X 3 ) ( OR =2.921, 95% CI : 1.114-7.662, P =0.029) were independent risk factors for VTE in patients with liver failure after artificial liver support therapy. The equation for individual predicted probability was P =1/[1+e -(7.425-1.384X 1 -2.103X 2 +1.072X 3 ) ]. The ROC curve analysis showed that Caprini score had an area under the ROC curve of 0.802 (95% CI : 0.721-0.882, P < 0.001), and the multivariate model had an area under the ROC curve of 0.768 (95% CI : 0.685-0.851, P < 0.001), while the combination of Caprini score and the multivariate model had an area under the ROC curve of 0.957 (95% CI : 0.930-0.984, P < 0.001). Conclusion The Caprini risk assessment model has a high predictive efficiency for the risk of VTE in patients with liver failure after artificial liver support therapy, and its combination with the multivariate predictive model can significantly improve the prediction of VTE.