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2018 Vol. 48, No. 2 Published: 2018-02-25   79 Preparation and in vitro evaluation of nano-scal polymer micelles simultaneously encapsulating disulfiram and doxorubicin Xu Xiafang, Tong Mengqi, Jiang Qianyao, Zhao Yingzheng, Xu Helin. Objective: To prepare the nano-scal polymer micelles simultaneously encapsulating disulfiram (DSF) and doxorubicin (DOX) and to evaluate its physical and chemical properties in vitro and the ability to reverse the multi-drugs resistance of breast cancer cells against doxorubicin. Methods: Using monomethoxy (polyethylene glycol)-b-P (D, L-lactic-co-glycolic acid)-b-P (L-glutamic acid) (mPEG-PLGA-PGA) as materials, the dual-drugs loaded polymer micelles (DSFDOX-NPs) was prepared by dialysis method and its physical and chemical properties were studied by dynamic light scattering (DLS), transmission electron microscope (TEM). Drug loading content (DLC) and drug loading efficiency (DLE) were measured by HPLC. Furthermore, in vitro drug release profiles from DSFDOX-NPs were explored by dynamic dialysis method. Finally, the cellular toxicity against MCF-7/ADR and the cellular uptake of DSFDOX-NPs were evaluated. Results: DSFDOX-NPs had a mean diameter of (107.5±0.2) nm, and Zeta potential of (-13.7±0.1) mV, determined by DLS. TEM image showed that DSFDOX-NPs had spherical shape, and dry mean diameter of 95 nm. DLC and DLE of disulfiram was 1.91% and 80.30% respectively, while DLC and DLE of doxorubicin was 2.28% and 96.2% respectively. The sustained-release of both of DSF and DOX from DSFDOX-NPs was observed, exhibiting a cascade release profiles with the faster release of DSF followed by a slower release of DOX. DSFDOX-NPs can enhance the cellular uptake of DOX by drug-resistant MCF-7/ADR cells through inhibition of P-gP, displaying the significantly higher cytotoxicity against MCF-7/ADR than DOX solution. Conclusion: DSF and DOX is efficiently encapsulated in DSFDOX-NPs with a particle size of less than 100 nm. The sustained-release of DSF and DOX from DSFDOX-NPs is also observed. More importantly, DSFDOX-NPs displayed multidrugresistance reversal effect on MCF-7/ADR cell. 2018 Vol. 48 (2): 79-85 [Abstract] ( 723 ) HTML (1 KB)  PDF (1610 KB)  ( 1211 ) 86 Effects of neuromuscular electrical stimulation on lower muscle of hypoxia-hypercapnia rats via regulation of miR1-related signaling pathway PAN Lulu, ZHI Yinghao, WANG Xiaotong Objective: To explore the effects of neuromuscular electrical stimulation on gastrocnemius muscle of chronic intermittent hypoxia-hypercapnia (CIHH) rats and try to find out how much the miR-1 related signaling pathway is involved in. Methods: Twenty four male SD rats were randomly divided into three groups: the normal control group (NC), hypoxia-hypercapnia group (HH), hypoxia-hypercapnia+electrical stimulation group (HE). The HH and HE were placed in a chamber with hypoxia-hypercapnia for 8 h/d, 4 weeks in all. After 2 weeks, 30 min, 2 Hz and 100 Hz, 1:1 cyclic alternating pattern of electrical stimulation was applied in the HE, which lasted 2 weeks. The NC was only tied up and placed with electrode plates. The endurance running capacity was assessed by the treadmill. The muscle phenotype was observed by immunohistochemical staining. The expression of miR-1 by qRT-PCR was detected. The expression of HDAC4 (histone deacetylase 4), MEF2 (myocyte enhancer factor 2) and PGC-1α were detected by Western blot. Results: After 4 weeks, the HH group exhibited a decreased MHCI-to-MHCII shift (P<0.05). Nuclear HDAC4, MEF2 and PGC-1α was also significantly decreased (P<0.05). In contrast, miR-1 was significantly increased (P<0.05). Electrical stimulation can partly reversed those changes, and improved endurance running capacity. Conclusion: Our study indicate the miR-1 related signaling pathway may play a role in the response of CIHH-impaired muscle to changes during electrical stimulation. 2018 Vol. 48 (2): 86-90 [Abstract] ( 810 ) HTML (1 KB)  PDF (1538 KB)  ( 914 ) 91 Effect of long noncoding RNA CasC7 on proliferation of cervical cancer cells AN Dewen, HU Yingying, LIANG Zongwen, XIANG Junmiao, YU Danyang, DUAN Ping Objective: To investigate the expression level of lncRNA CasC7 in cervical cancer and its role in tumorigenesis and development of cervical cancer. Methods: Bioinformatics was utilized to analyse the expression level of lncRNA CasC7 in GSE9750, a gene expression microarrays for cervical cancer. Quantitative real-time PCR (qRT-PCR) was used to detect the expression level of CasC7 in cervical cancer tissues, cervical intraepithelial neoplasia tissues, normal cervical tissues and normal cervical epithelial cells (H8 cells), cervical cancer cells (SiHa cells, CaSki cells and HeLa cells). Association between the expression of CasC7 and clinicopathological features of cervical cancer was analyzed. Meanwhile, the CCK-8 assays was used to measured cell proliferation when CasC7 expression was inhibited. Results: LncRNA expression in cervical cancer and cervical intraepithelial neoplasia was higher than that in normal cervical tissues (P<0.01). Compared with normal cervical epithelial cells, CasC7 was up-regulated in SiHa cells, CaSki cells and HeLa cells (P<0.05). High expression of CasC7 was correlated with advanced FIGO stage (P=0.001) and poor differentiation (P=0.015). However, neither patients age nor tumor size was associated with the expression of CasC7 (P>0.05). CCK-8 assays showed that cell proliferation of SiHa cells, CaSki cells and HeLa cells was suppressed by CasC7 knockdown (P<0.05). Conclusion: CasC7 may act as an oncogene during the development of cervical cancer. 2018 Vol. 48 (2): 91-95 [Abstract] ( 793 ) HTML (1 KB)  PDF (1505 KB)  ( 825 ) 96 Analysis of risk factors for coronary artery lesions in Kawasaki disease with delayed immunoglobulin treatment QIU Huixian, SHI Hongying, HE Yuee, RONG Xing, WU Rongzhou, CHU Maoping. Objective: To identify the risk factors for coronary artery lesions with intravenous immunoglobulin (IVIG) treatment of Kawasaki disease (KD) >10 days after onset at different status. Methods: Nine hundred and thirty patients diagnosed as KD were collected from January 2009 to December 2014 at the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University. Patients were divided into the delayed IVIG therapy group (who received IVIG treatment after 10 days) and the conventional therapy group (who received IVIG treatment within 10 days). Coronary artery lesions between the two groups were compared. Risk factors for coronary artery lesions were investigated. Results: In the delayed IVIG therapy group, the frequency of coronary artery lesions within 1 month was higher (42.4% vs. 17.7%, P<0.001). The interaction analysis indicated that delayed IVIG therapy children with higher level of C-reaction protein (CRP, >79 mg/L, OR=5.68, 95%CI: 1.17-27.59, P<0.05), erythrocyte sedimentation rate (ESR, >34 mm/h, OR=4.11, 95%CI: 1.62-10.46, P<0.05), neutrophils and aspartate transaminase had higher risk to develop coronary artery lesions. Delayed IVIG therapy with lower level of albumine, red blood cell also had higher risk to develop coronary artery lesions. Compared with controventional therapy group without changing of departement, the delayed therapy with changing of department had higher risk to develop coronary artery lesions (OR=7.29, 95%CI: 1.85-45.17, P=0.0327). Conclusion: Delayed IVIG treatment leads to increased risk of coronary artery lesions in KD children. Delayd IVIG treatment combined with higher level of CRP, ESR, neutrophils, aspartate transaminase, lower level of red blood cell and not admitted to cardiology department at initial time has higher risk to develop coronary artery lesions. 2018 Vol. 48 (2): 96-100 [Abstract] ( 722 ) HTML (1 KB)  PDF (1383 KB)  ( 802 ) 101 Effect of ZAG on lipid metabolism of colon adenocarcinoma cachexia mice Zhang Peichen, Shi Lingyan, Zhou Feng, Wu Wenyi, Yu Zhen Objective: To investigate the effect of zinc-alpha2-glycoprotein (ZAG) on lipid metabolism of mice with cancer cachexia (CC) using a mouse model of colon adenocarcinoma cachexia. Methods: A mouse model of colon adenocarcinoma cachexia was well established by inoculating mouse colon cancer cell line, colon 26 into BALB/c mice. A total of 18 BALB/c mice were evenly divided into 3 groups randomly: healthy control (HC) group, tumor control (TC) group and cancer cachexia (CC) group (n=6). Adjacent non-tumor tissues and adenocarcinoma of colon (AC) tissues isolated from TC group and CC group were collected to determine ZAG mRNA and protein level. ZAG protein concentration and mRNA level in subcutaneous adipose tissues of each group were determined, while the correlation between ZAG mRNA level and weight loss was confirmed. mRNA expression level of ZAG and adiponectin in serum were measured, and the correlation between them in TC group and CC group was analyzed. Finally, weight was recorded in cancer cachexitic model of mice with colon adenocarcinoma transplantation followed by adenovirus-constructed pcDNA or pcDNA-ZAG injection. Results: ZAG mRNA and protein level of AC tissues and adipose tissues in CC group was significantly higher than those in TC group (P<0.05). ZAG mRNA level was correlated positively with weight loss in adipose tissue. Besides, the serum mRNA levels of ZAG and adiponectin in CC group were significantly higher than those in TC group and were positively correlated. In addition, mice weight was observably reduced after treatment with pcDNA-ZAG (P<0.05). Conclusion: Enhanced ZAG can significantly promote the steatolysis, and affect the weight in colon adenocarcinoma cachexia mice. 2018 Vol. 48 (2): 101-105 [Abstract] ( 695 ) HTML (1 KB)  PDF (1624 KB)  ( 1049 ) 106 GC-MS method for determination of 16 kinds of PAES in drugs and its packaging materials LI Zhou, CAI Xiaojun, XIANG Zheng Objective: To investigate a GC-MS method for simultaneous determination of 16 kinds of PAEs in drugs and its packaging materials. Methods: PAEs was extracted from drugs and its packaging materials by solvents, such as hexane, using ultrasonic method. The samples were separated by gas chromatography. Meanwhile, the mass spectrometry was used for qualitative analysis and internal standard method was taken for quantitative analysis. Results: The standard curves for 16 kinds of PAEs were in good linear relationship in the range of 50-500 ng•mL-1, with average recovery being 99.85%-100.10% and RSD < 8.95%. Conclusion: This method is effective, accurate and highly sensitive for the rapid detection of PAEs in drugs and its packaging materials. 2018 Vol. 48 (2): 106-109 [Abstract] ( 684 ) HTML (1 KB)  PDF (1485 KB)  ( 1022 ) 110 The self-renewal ability of ganlioside GD2+ breast cancer stem cells via selectively suppressing STAT3 activation ZHU Lihe, HU Lufeng, JI Jianle, ZHU Wei Objective: To investigate the effect of selectively inhibiting STAT3 on the self-renewal ability of GD2+ breast cancer stem cells by stattic. Methods: The GD2+ breast cancer stem cells were sorted out from human breast cancer cell lines MDA-MB-231 via flow Cytometry. The obtained GD2+ stem cells were identified. Western blot was applied to evaluate the expression of STAT3, pSTAT3 protein. Stattic inhibition test was applied to explore the effect of inhibiting STAT3 on the self-renewal ability of GD2+ breast cancer stem cells. Results: GD2+ breast cancer stem cells grew as spheres cultured in serum-free suspension. The mammpsphere formation number of GD2+ breast cancer stem cells was (22.38±3.44) per 1 000 cells, significantly higher than that of GD2-cells (7.14±1.14) (P<0.05). The ammpsphere size of GD2+ cells was (148.47±41.34) μm, significantly larger than that of GD2- cells (57.10±13.58) μm (P<0.05). Beginning from the second day, GD2+ breast cancer stem cells were less proliferating than GD2- cells (P<0.05). The expression of pSTAT3 protein in GD2+ breast cancer cells was significantly higher than that in GD2- breast cancer cells (P<0.05), but there was no significant difference in the expression of STAT3 protein between the two groups (P>0.05). After the intervention of selective STAT3 inhibitor Stattic on GD2+ breast cancer stem cells, the proliferation and mammpsphere forming ability of breast cancer stem cells were decreased (P<0.05). Conclusion: The selective STAT3 inhibitor Stattic can inhibit the self-renewal ability of GD2+ breast cancer stem cells. 2018 Vol. 48 (2): 110-114 [Abstract] ( 721 ) HTML (1 KB)  PDF (1556 KB)  ( 903 ) 115 Correlation between preeclampsia and second-trimester maternal serum levels of AFP and free β-HCG#br# CHEN Yiming, LU Sha, HE Pei, GU Linyuan, ZHANG Yanzhen, CHU Xuelian, LIAN Jiejing, ZHANG Min. Objective: To determine if second-trimester maternal serum alpha-fetoprotein (AFP) level and free β-HCG level can predict preeclampsia. Methods: Second-trimester maternal serum levels of AFP and β-HCG in pregnancies who delivered in Hangzhou Women’s Hospital from January 2015 to August 2016 were analyzed. A prospective case-control study included 55 mild preeclampsia patients, 36 severe preeclampsia patients and 100 control pregnant women. AFP and free β-HCG among groups were compared. The correlation of age, weight, gestational age, AFP, free β-HCG and preeclampsia, severe preeclampsia were analyzed by unconditional logistic regression analysis. The diagnostic efficacy of AFP and free β-HCG were evaluated by ROC curves. Results: AFP of mild preeclampsia group and severe preeclampsia group was significantly higher than control group (P<0.01). Logistic regression analysis showed that AFP was independent factor of mild and severe preeclampsia (P<0.05), but it has lower sensitivity. Conclusion: Higher second trimester maternal serum AFP level is a risk factor for preeclampsia. It can serve as a screening test for preeclampsia. However, single detection of AFP test is limited as the lower sensitivity. Combining other markers will make it more sufficient and reliable. 2018 Vol. 48 (2): 115-118 [Abstract] ( 677 ) HTML (1 KB)  PDF (1420 KB)  ( 966 ) 119 Network meta-analysis of the efficacy of traditional Chinese medicine injection combined with mesalazine in the treatment of active ulcerative colitis Feng Jinshan, Li Jinyu, Yang Yuee, Chen Xiuyan, Yang Zheng Objective: To compare the efficacy of 6 kinds of traditional Chinese medicine injections (TCMi) combined with mesalazine in the treatment of active ulcerative colitis. Methods: Randomized controlled trials (RCTs) of TCMi were searched in domestic and foreign databases in accordance with the predefined inclusion and exclusion criteria. Data process and analysis were performed with WinBUGS and Stata softwares. Results: A total of 561 patients were enrolled in 6 RCTs in the present study. Seven treatment groups included 6 TCMi-combined-with-mesalazines and 1 group used mesalazine alone. Differences in efficacy among the 7 treatment groups did not have statistical significance. Conclusion: There is no statistical difference in the efficacy of 6 TCMi combined with mesalazine in the treatment of active ulcerative colitis. 2018 Vol. 48 (2): 119-122,127 [Abstract] ( 600 ) HTML (1 KB)  PDF (2588 KB)  ( 934 ) 123 Study on lumbar disc annulus fibrosus by MR diffusion tensor imaging YAN Chunli, ZHOU Min, LI Mingying, CHEN Xingcan Objective: To evaluate the manifested ability of MR diffusion tensor imaging for lumbar disc annulus fibrosus. Methods: 15 healthy volunteers (control group) and 20 patients (patients group) with lumbar intervertebral disc herniation (LDH) underwent sagittal T1WI, T2WI and axial T2WI, DTI scan. Fiber tracking (FT) maps of these lumbar intervertebral discs were reconstructed based on the DTI data. The classification criterion of FT map was set and all FT maps  were classified as regulars [continuous (A1) and discontinuous (A2)], disordes [continuous (B1) and discontinuous (B2)] and masses (C) by the morphological characteristics and continuity of annulus fibrosus in FT maps. All lumbar intervertebral discs were evaluated through Pfirrmann’s classification system. Spearman’s correlation coefficient of data was analyzed by SPSS software between the types of FT maps and Pfirrmann’s classification system. Results: There were 74 lumbar intervertebral discs examined in control group, in which type A1 was 22 (grade I 10, II 12), type A2 was 32 (grade I 14, II 18), type B1 were 15 (grade I 1, II 6, III 8), type B2 were 3 (grade II 1, III 2), type C were 2 (grade III 2); there were 86 lumbar intervertebral discs examined in patients group, in which, type A1 was 8 (grade II 5, III 3), type A2 was 7 (grade II 4, III 3), type B1 were 32 (grade II 2, III 27, IV 3), type B2 were 4 (grade III 4), type C were 35 (grade III 16, IV 17, V 2). 22 LDH in 20 patients underwent surgical operation. And there were 9 contained LDH (B1 2, B2 3, C 4) and 13 noncontained LDH (B1 2, C 11) were confirmed by operation. There is strong correlation between the type of FT maps and Pfirrmann’s classification system (r=0.741, P<0.001). Conclusion: DTI can manifest degeneration of lumbar intervertebral disc noninvasively. The pattern of FT map for lumbar disc annulus fibrosus plays a guiding role in clinical treatment. 2018 Vol. 48 (2): 123-127 [Abstract] ( 682 ) HTML (1 KB)  PDF (1616 KB)  ( 902 ) 128 Analysis of related factors of deep venous thrombosis in lower extremity in perioperative period of rectal cancer ZHENG Xiangjian, XU Jianmin. Objective: To analyze the related factors of deep venous thrombosis in lower extremity in perioperative period of rectal cancer. Methods: Three hundred and ninety-three patients treated by rectal-cancer surgery in our hospital from October 2013 to October 2016 were reviewed retrospectively. The clinical data including age, gender, hospital stay, operation time, operation method, occupation, hypertension, coronary heart disease, diabetes, BMI, smoking status, drinking status, postoperative exercises and treatment by postoperative elastic stockings or intermittent pneumatic calf compression were collected and quantitative analysis was made. Results: In this study, 59 patients were included in the DVT group and 334 patients in the Non-DVT group. There was signifcant difference between the two groups (P<0.05) in age (>60 years old), hypertension, occupation type, operation time, smoking status, postoperative exercises, the treatment by postoperative elastic stockings or intermittent pneumatic calf compression. The multivariate analysis showed that age, smoking, hypertension and operation time were independent risk factors (P<0.05). Physical labour, treatment by postoperativeexercises or postoperative elastic stockings and intermittent pneumatic calf compression were protective factors for DVT (P<0.05). Conclusion: Rectal cancer patients with older age (>60 years old), hypertension and longer operation time should pay much attention to the prevention of DVT in perioperative period, and postoperative exercises can be a precautionary measures. 2018 Vol. 48 (2): 128-130,134 [Abstract] ( 689 ) HTML (1 KB)  PDF (2458 KB)  ( 856 ) 131 Emergence agitation in pediatric patients following desflurane or sevoflurane anesthesia YE Xuefei, HUANG Wenguang, QIN Peishun, LIN Han, LIAN Qingquan. Objective: To compare the emergence agitation in pediatric patients undergoing minor ENT surgery under sevoflurane or desflurane anesthesia. Methods: One hundred children of 1-7 years old undergoing minor ear-nose-throat (ENT) surgery were randomly divided into two groups: 50 cases in sevoflurane group (S group) and 50 cases in desflurane group (D group). After inhaled 8% sevoflurane combined 70% nitrous oxide with 10 L/min fresh gas, an IV was established and fentanyl 2 μg/kg followed by rocuronium 0.4 mg/kg was administered to facilitate endotracheal intubation. For maintenance of anesthesia, children were randomly assigned to receive either sevoflurane (1.0±0.2 ) MAC or desflurane (1.0±0.2) MAC, with a constant fresh gas flow of 2 L/minnute of 50% nitrous oxide in oxygen. The HR, MAP, SPO2 of admission to OR (T1), during the operation (T2), after extubation (T3) were recorded. Besides, degree of agitation using PAED scale, modified Aldrete scores and CHIPPS scores on admission to PACU were all recorded and followed every 10 minutes. Results: PAED scores in D group were significantly lower than S group at admission and 10 minnutes after PACU (P<0.01). Modified Aldrete scores in D group were also significantly higer than S group at admission and 10 minnutes in PACU (P<0.01, P<0.05). Besides, CHIPPS pain score in D group was significantly lower than S group when arriving in PACU (P<0.01). Conclusion: The emergence agitation associated with the use of desflurane is lower than sevoflurane anesthesia in children undergoing minor ENT surgery. 2018 Vol. 48 (2): 131-134 [Abstract] ( 689 ) HTML (1 KB)  PDF (1380 KB)  ( 995 ) 135 Treatment of varicose veins of lower extremity by endovenous laser combined with foam sclerotherapy in ultrasonic positioning and swelling anesthesia MEI Tong, XU Tingting, HUANG Weijin, CHEN Jinhui, TANG Tao, LU Yongming.  Objective: To evaluate the efficacy and safety of endovenous laser treatment combined with foam seleretherapy for varicose veins of the lower limbs. Methods: According to the standard of varicose veins of the lower limbs, 79 patients (93 limbs) were selected. The trunk of great saphenous vein was treated with EVLT in ultrasonic positioning and swelling anesthesia and then sclerosing foam was injected into the tributary of the GSV. Ultrasound venous reflux was evaluated and the related postoperative complications after one week was reviewed 1 month, 3-month and 6-month postoperative was followed-up. Results: There was different degree of decline in the CEAP classification and no obvious pain was found in all patients after one postoperative week. There were no lower extremity deep venous thrombos, pulmonary embolism and other serious complications. There were 4 cases of superficial thrombophlebitis, 3 cases of pigmentation, 2 cases of saphenous nerve injury.There were no residual and recurrence of varicose in postoperative 3-month and 6-month follow-up. Conclusion: The treatment of varicose veins of lower extremity by endovenous laser combined with foam sclerotherapy is a safe and effective minimally invasive surgical procedure in ultrasonic positioning and swelling anesthesia. 2018 Vol. 48 (2): 135-137 [Abstract] ( 719 ) HTML (1 KB)  PDF (1374 KB)  ( 863 ) 138 Analysis of the performance and influencing factors in preterm infants at first oral feeding stage LIU Jiaqi, ZHANG Kunhua, ZHANG Shuanghong, ZHANG Liping  Objective: To investigate the oral feeding performance in preterm infants at first oral feeding stage and analyze the influencing factors. Methods: One hundred and two premature infants at first oral feeding stages were evaluated, the basic data, the feeding milk transfer rates and the feeding proficiency were collected. Preparation behavior and behavior status in feeding, the volume of oral intaked milk within 5 minutes and the feeding time were recorded. Feeding velocity and feeding efficiency were calculated. Results: The feeding performance of preterm infants in the different gestational ages, birth weights and neonatal medical index showed significant differences (P<0.05). Multifactor analysis results showed that gestational age (OR= 3.650, 95%CI=1.123-4.531, P<0.001), birth weight (OR=3.104, 95%CI=1.207-4.301, P<0.001), sucking reflex (OR=2.031, 95%CI=1.150-3.541, P=0.011) and neonatal medical index (OR=0.612, 95%CI=0.371-0.819, P= 0.015) were influencing factors. Conclusion: Based on influencing factors, clinical nurses should strengthen the management of oral feeding in the preterm infants. 2018 Vol. 48 (2): 138-141 [Abstract] ( 946 ) HTML (1 KB)  PDF (1405 KB)  ( 1031 ) 142 Clinical application of diversified nursing care on self management in patients with type 2 diabetes mellitus#br# Hong Yirong, QU Jia, HUANG Fang. Objective: To investigate the clinical application of the diversified nursing in type 2 diabetes patients’ self health management. Methods: Two hundred patients with type 2 diabetes were randomly divided into two groups, the control group received routine hospital diabetes care, and the experimental group received diverse forms of nursing. The self-management behavior scale, fasting blood glucose, 2 hours postprandial blood glucose and glycated hemoglobin levels in 2 groups were compared. Results: After intervention, the score of self-management behavior in the experimental group was significantly higher than those before intervention, and also significantly higher than those of the control group. Fasting blood glucose, postprandial 2 hours blood glucose and glycosylated hemoglobin in the experimental group were significantly decreased after intervention, and significantly lower than that of the control group. Conclusion: Diversified nursing care can improve the self-management ability of patients with type 2 diabetes mellitus, and can maintain good blood glucose level. 2018 Vol. 48 (2): 142-144 [Abstract] ( 668 ) HTML (1 KB)  PDF (1398 KB)  ( 965 )

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