Objective To constract and evaluate the cationic anticancer peptide Temporin-1CEa liposomes and evaluate anti-breast cancer activity in vitro.Methods The polyethylene glycol (PEG)-modified liposomes containing Temporin-1CEa, one recently discovered cationic anticancer peptide ( CAP) , were constructed by using reverse-phase evaporation method.The encapsulation efficiency, particle size and Zeta potential of the Temporin-1CEa-containing liposomes (Temporin-1CEa-LIP) were characterized.In addition, that had the furhter evaluated of the stability and specific toxicity against human breast cancer MCF-7 cells in vitro.Results The data suggested that the PEG-modified liposomes served a promising drug delivery system for CAPs, those indicated by the encapsulation efficiency was (55.57 ±1.56)%, the particle size was (105.3 ±1.37) nm and the Zeta potential was ( -16.17 ±0.964) mV.Moreover, the in vitro test also indicated that Temporin-1CEa-LIP exerted good stability in serum, and it could be efficiently uptaken by MCF-7 cells.Most importantly, after 24h exposure, Temporin-1CEa-LIP showed toxicity against MCF-7 cells, as potent as Temporin-1CEa. Conclusion The results demonstrates that the PEG-modified liposome is a good drug-delivery system and Temporin-1CEa-LIP could serve as potential anti-tumor candidate for cancer therapy.

AIMTo study the chemical composition of Tripterygirm wilfordii Hook. f.

METHODSColunm chromatography was used to separate the chemical constituents. UV, IR, MS, HRMS, 1HNMR, 13CNMR (COM and OFR), 1H-1H COSY, 1H-13C COSY, NOESY and COLOC spectra were used to determine the structures of the isolated constituents.

RESULTSTwo sesquiterpene alkaloids were isolated and their structures were elucidated as wilforgine and wilfordlongine on the basis of spectral evidence.

CONCLUSIONWilfordclonine is a new sesquiterpene alkaloid.

Lactones ; chemistry ; isolation & purification ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Pyridines ; chemistry ; isolation & purification ; Tripterygium ; chemistry

OBJECTIVETo construct the KbvR gene of LuxR family deletion mutant and complementation strains from Klebsiella pneumoniae NTUH-K2044 and analyze the effect of KbvR on bacterial growth, biofilm formation and capsular synthesis.

METHODSA KbvR gene deletion mutant strain was constructed using the suicide vector pKO3-Km, and the gene fragment including KbvR coding region, promoter area and transcription termination area were amplified and cloned into pGEM-T-easy plasmid to construct KbvR complementation strain. The growth curves of the wild-type strain, KbvR gene deletion mutant strain and complementation strain were observed to assess the effect of KbvR on bacterial growth. Crystal violet staining method was used to measure the effect of KbvR on biofilm formation; the effect of KbvR on capsular synthesis was detected using string test, centrifugal test and RT-PCR.

RESULTSThe KbvR deletion mutant and complementation strains were constructed successfully. KbvR gene did not affect the growth of the bacteria, but biofilm formation and capsular synthesis were attenuated in KbvR deletion mutant strain.

CONCLUSIONAs a transcription factor of the LuxR family orphans of the quorum sensing system, KbvR positively regulates bacterial biofilm formation by affecting capsular synthesis.

Bacterial Capsules ; Bacterial Proteins ; metabolism ; Biofilms ; Gene Deletion ; Klebsiella pneumoniae ; cytology ; Plasmids ; Promoter Regions, Genetic

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

OBJECTIVETo describe the distribution changes of the mortality rate for cervical cancer in China between the 1970's and 1990's and provide the scientific evidence for the prevention and control strategies for cervical cancer campaign in China between next century.

METHODSData from two National Surveys for the Causes of Death in 1970's and 1990's in China. The crude and adjusted mortality rates for the cervical cancer and the distributions based on age and area were calculated and described. The comparison of the differences of changes between two mortality rates periods and together with its trends were shown based on the age-standardized.

RESULTSDuring two decades, the mortality rate for cervical cancer was 10.7 per 100,000 in 1970's which declined to 3.89 per 100,000 in 1990's, and from the 3rd ranking among all female malignant tumors to the 6th in 1990's (decreased about 63.64%). But the declination was not evenly. There have still been some high-risk areas, most located in rural countries in the mid-west of China, with rates remain unchanged and even at the highest level in the world, such as Wudu in Gansu and Yangcheng in Shanxi. A big difference was showed between rural country and city, but in both of them, the mortality rates in 1990's were significantly much lower than in 1970's (P = 0.001) at each five-year age group. And in the city, there was a much sharper increased trend in young women in 1990's.

CONCLUSIONSThe mortality rate for cervical cancer campaign in China has been substantially declined during past twenty years, but it's still a major health problem for women, especially in rural China. The focus of the prevention and control for the cervical cancer in the next century should put on rural areas, especially in mid-west of China and young women in the city.

Adult ; Age Factors ; Aged ; Cause of Death ; trends ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Retrospective Studies ; Rural Health ; Uterine Cervical Neoplasms ; mortality ; prevention & control

·AMI:To evaluate the clinical efficacy of the segmented bifocal intraocular lens (MIOL) SBL-3. ·METHODS:Totally 26 cases (26 eyes) of age-related cataract who received phacoemulsification and implantation of MIOL SBL-3 were enrolled from February 2016 to June 2017 in our hospital as the SBL-3 group. And 28 cases (28 eyes) of age-related cataract who received phacoemulsification and implantation of single focus intraocular lens intraocular lens TecnisZA9003 were enrolled as control group. At postoperative 3mo, the uncorrected visual acuity, corrected visual acuity, contrast sensitivity and patient satisfaction were compared between two groups. ·RESULTS: At preoperative, there was no statistical difference in uncorrected distance, intermediate and near visual acuity between two groups (P > 0. 05). At postoperatively 3mo, the uncorrected distance, intermediate and near visual acuity in patients of both groups were significantly improved (P < 0. 05). At postoperative 3mo,the uncorrected intermediate and near visual acuity, distance-corrected intermediate and near visual acuity, contrast sensitivity at different spatial frequencies (3, 6, 12, 18c/d) and patient satisfaction in SBL- 3 group were significantly better than those of control group (P<0.05). ·CONCLUSION:The MIOL SBL-3 not only could provide preferable distance visual acuity, but also could provide better intermediate visual acuity, near visual acuity and contrast sensitivity,and greatly increase the visual quality and satisfaction at postoperative in cataract patients.

·AIM: To compare the visual quality after different multifocal lens (MIOL) implantation in cataract patients. ·METHODS: Totally 78 cases (78 eyes) of cataract patients who receiving phacoemulsification and implantation of different MIOL were enrolled from February 2016 to June 2017 in our hospital. According to the different type of implanted MIOL, the 78 cases (78 eyes) were divided Group A,B and C,each group was 26 cases (26 eyes). Patients in the Group A were implanted regional refraction MIOL SBL-3; patients in the Group B implanted step progressive diffraction MIOL SN6AD1;patients in the Group C implanted all - optical plane diffraction MIOL Tecnis ZMB00. The visual acuity, contrast sensitivity,defocus testing,contrast visual acuity and satisfaction of visual quality were compared in 3 groups at postoperative 3mo. · RESULTS: There was no statistically significant difference in postoperative adverse reactions between the three groups (P>0.05). There was statistical difference in uncorrected intermediate visual acuity, uncorrected near visual acuity, distance - corrected intermediate visual acuity and distance- corrected near visual acuity in 3 groups (P < 0. 05), and those in the Group C were significantly worse than those of Group A and B (P<0.05). There was statistical difference in contrast sensitivity at four spatial frequencies (3,6,12,18c/d) in light and light glare and three spatial frequencies (3, 6, 12c/d) in dark and dark glare in 3 groups (P<0.05), and those in the Group A were significantly better than those of Group B and C (P<0.05). According to the defocus testing of 3 groups,the visual acuity at -1.5D,-2.0D and-2. 5D in Group A and B was significantly higher than Group C (P<0. 05). There was statistical difference in contrast visual acuity at 20% and 9% of contrast ratio in 3 groups (P<0.05),and those in Group A were significantly better than Group B and C(P<0.05). There was statistical difference in visual quality satisfaction in 3 groups (P<0.05), and that in the Group A was significantly better than that of Group B and C (P<0.05). ·CONCLUSION:The region refraction MIOL SBL-3 not only could provide better distant, intermediate and near visual acuity, but also could provide better contrast sensitivity and contrast visual acuity, thereby greatly increase visual quality satisfaction.

BACKGROUND: Multilevel thoracolumbar fractures are mainly treated with percutaneous pedicle screw and open pedicle screw system, but the treatment effect of different systems and the accuracy rate of screw placement are controversial, resulting in the lack of uniform standards for choosing the treatment method. OBJECTIVE: To evaluate the effect of percutaneous pedicle screw and open pedicle screw system in the treatment of multilevel thoracolumbar fractures and to evaluate the accuracy of the screw placement. METHODS: Totally 90 patients with multilevel thoracolumbar fractures were divided into open pedicle screw group (n=43 cases) and percutaneous pedicle screw group (n=47) according to different surgical methods. Open pedicle screw group was treated with open pedicle screw treatment, and percutaneous pedicle screw group was treated with percutaneous pedicle screw. Comprehensive effects were analyzed by comparing perioperative indicators (operation time, postoperative drainage volume, and incision length) imaging index (anterior vertebral height percentage, posterior vertebral height percentage, sagittal Cobb angle), postoperative complications, and pedicle screw accuracy. RESULTS AND CONCLUSION: (1) The amount of bleeding, postoperative drainage volume, and incision length were less (shorter) in the percutaneous pedicle screw group compared with the open pedicle screw group (P < 0.05). However, operation time and the number of undergoing fluoroscopy were longer (more) in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). (2) Anterior vertebral height percentage and posterior vertebral height percentage were higher in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). Sagittal Cobb angle was smaller in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). (3) At 2 months after surgery, the complication rate was significantly lower in the percutaneous pedicle screw group (4%) than in the open pedicle screw group (14%) (P < 0.05). (4) The accuracy rate of pedicle screw was significantly higher in the percutaneous pedicle screw group (92.1%; 279 screws) than in the open pedicle screw group (77.0%; 257 screws) (P < 0.05). (5) Results indicated that percutaneous pedicle screw fixation is characterized by less trauma and rapid recovery in the treatment of multilevel thoracolumbar fractures. It is helpful for the reduction of the injured vertebra, the maintenance of vertebral height; the safety and the accuracy of screw placement are high.

In order to differently diagnose avian influenza virus (AIV) subtypes, the HA gene of AIV H9 subtype was cloned, expressed and utilized in an enzyme-linked immunoad sorbent assay (ELISA). HA gene (1683bp) of H9N2 AIV was amplified by RT-PCR from a strain of field isolated H9N2 AIV, and its identity was confirmed by sequencing. The HA gene was subcloned into prokaryotic expression vector pGEX-KG with its secretion signal sequence removed. The expressed HA-GST fusion protein in E. coli BL21 was characterized by SDS-PAGE and western blotting analysis as a 90kD protein with immunogenicity. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation. The HA-GST fusion protein was used to establish an indirect ELISA for the detection of antibodies to H9 subtypes of AIV. The assay has 91.57% specificity to H9 AIV, 92.31% sensitivity and excellent reduplication. It could be used to differently detect antibodies to H9 AIV.
Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; Humans ; Influenza A Virus, H9N2 Subtype ; genetics ; Influenza, Human ; diagnosis ; virology ; Recombinant Proteins ; biosynthesis ; genetics

AIM:To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS:The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. Af-ter treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS:AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-Ⅰ were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION:AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.