Adenocarcinoma, Clear Cell ; pathology ; Female ; Humans ; Middle Aged ; Ovarian Neoplasms ; pathology

AIMTo investigate factors affecting the properties of antisense oligodeoxy nucleotides (ASON)-liposomes complex and their cellular uptake.

METHODSThree types of blank liposomes were prepared by reverse-phase evaporation vesicles, and the complex were obtained through physical absorption. The light microscope was used to observe morphology characteristics of the complex. Drug loading capacity was analyzed by agarose gel electrophoresis. The transfected cell percentage and means fluorescence intensity were determined by flow cytometric analysis using M3 myeloma cell as a model.

RESULTSThe neutral liposome showed no aggregation while the cationic liposomes appeared some different extent aggregation in different medium when associated antisense oligodeoxynucleotides. The drug loading capacity depended on the ratio of +/- and the cationic charge density on the lipid membrane. The two kinds of cationic liposomes appeared different principles of loading ASON. As far as cellular uptake, The neutral liposomes showed no improvement of cellular uptake of ASON. However, the cationic liposomes were shown to enhance the cellular uptake of ASON if the appropriate +/- charge ratio was used. The optimal cellular uptake was achieved when +/- charge ratio was at 0.5:1 and 1:1 for SA-I liposome and SA-II liposomes, respectively.

CONCLUSIONThe cationic liposomes improved the loading capacity and cell uptake of antisense oligodeoxynucleotides, which was determined by +/- charge ratio and charge density.

Amines ; metabolism ; Biological Transport ; Drug Carriers ; Drug Delivery Systems ; Humans ; Liposomes ; pharmacokinetics ; Multiple Myeloma ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; pharmacokinetics ; Random Allocation ; Tumor Cells, Cultured ; metabolism

OBJECTIVE: To investigate a nwe, simple technique for preparation of interferon-alpha-liposomes, which may be suitable for industrial use. METHODS The uniform design coupled with computerized optimization was utilized to screen the formulation and preparation procedure of interferon-alpha-liposomes. Pro-liposomes were prepared by the powder bed grinding method and combined with interferon-alpha-solution to form interferon-alpha-liposomes. Liposome size was determined by the particle size analyzer. Free interferon-alpha and interferon-alpha-liposome were separated by gel filtration. Then the recovered activity of interferon-alpha was analyzed by enzyme-linked immunosorbent assay. RESULTS The result demonstrated that the best interferon-alpha-liposome formulation was as follows: the protectant was sorbitol; weight ratio of protectant to lipid was 5:1; weight ratio of octadecytamin to lipid was 1:9; weight ratio of sobey phosphatidylcholine to cholesterol was 9:1 respectively. Interferon-alpha-liposome size determined by the particle size analyzer was 80.8+/-36 nm and the encapsulation efficiency was 59.0+/-3.3%. CONCLUSION The powder bed grinding method can be used to prepare pro-liposomes which can be easily combined with interferon-alpha-solution to form interferon-alpha-liposomes.

Objective: To investigate the incidence of CMV infection(CMV-I) and CMV related diseases (CMV-D) after allogeneic hematopoietic stem cells transplantation in 70 consecutive allogeneic hematopoietic stem cells transplantation(allo-HSCT) patients and to search for the optimal prophylactic strategy.Methods: Blood samples were monitored using the CMV pp65 antigenemia assay.Of the 70 patients observed,30 patients with chronic myeloid leukemia［CML:CP(27),AP(2),BC(1)］,12 with acute myeloblastic leukemia(AML),10 with acute lymphoblastic leukemia(ALL)and other cases were NHL(3), AA(5), MDS(7), SCLC with pancytopenia (1),CLL(1), and MF (1). Sixty six patients received HLA - identical siblings transplantation and four received tranplants from their HLA- haploidentical donors. Seventy cases included allo-PBPCT (64 cases) , allo-BMT (4 cases) and allo-PB+BMT (2). Before transplantation, all patients and donors received CMV serological examination except 4 pairs of donors/recepients. All 66 patients (3 cases were CMV IgM positive) and 64/66 donors were CMV IgG positive. Results：After transplantation, 64/70 patients developed CMV viremia during monitoring period. Forty three of 70 patients developed CMV-D.Thirty five of them suffered from CMV-associated interstitial pneumonia(CMV-IP). The high peak levels of CMV antigenemia were associated with development of CMV disease　. Close correlation was found between acute graft vs host disease(GVHD) and CMV disease. The patients were followed up for 2 to 24 months. The patients who received preemptive therapy(group A)had significantly better outcome than CMV disease group(group B, P=0.0001). Conclusions: The results suggest that CMV antigenemia has high predictive value for subsequent CMV disease and CMV pp65 antigenemia -guided early therapy has particular advantage for avoiding morbidity and mortality caused by CMV disease.

OBJECTIVETo investigate the expression of the Pim-1 gene in the LNCaP cells of the animal model of orthotopically implanted prostate cancer by surgical castration simulating androgen-deprivation therapy.

METHODSWe equally allocated 32 male BALBc-nu mice into 4 groups, androgen-dependent prostate cancer (ADPC), androgen-deprivation therapy (ADT) , castration-resistant prostate cancer (CRPC) and blank control, and established the models of orthotopically implanted tumor using human prostate cancer LNCaP cells. We detected and ,compared the expressions of Pim-1, PSA, and androgen receptor (AR) in the tumor tissues of different groups by RT-PCR. qRT-PCR, ELSIA and immunohistochemistry.

RESULTSThe relative gray scales in the ADPC and CRPC groups were 0.59 ± 0.01 and 1.14 ± 0.02, with statistically significant differences from 0.62 ± 0.03 in the ADT group (P < 0.05), and the Δ Ct values of Pim-1 were 6.15 ± 0.34 and 4.56 ± 0.23 in the former two groups, also with significant differences from 5.11 ± 0.21 in the latter (P < 0.05). The results of 2-ΔΔ Ct relative quantification analysis showed that the amplification products of Pim-1 in the ADT and CRPC groups increased 2.05 and 3.01 times respectively that of the ADPC group. The concentration of PSA was significantly higher in the ADPC ([480 ± 25] pg/ml) and CRPC ([870 ± 23] pg/ml) than in the ADT ([170 ± 32] pg/ml) and blank control groups (0 µg/L) (P < 0.01). The mean optical densities of Pim-1 and AR proteins were 0.017 ± 0.002 and 0.032 ± 0.009 in the ADPC group and 0.024 ± 0.002 and 0.040 ± 0.011 in the CRPC group, both with significant differences from those in the ADT group (0.018 ± 0.001 and 0.019 ± 0.006) (P < 0.01).

CONCLUSIONPim-1 is highly expressed in nude mice with prostate cancer receiving androgen-deprivation therapy and plays an important role in the progression and metastasis of prostate cancer.

Androgen Antagonists ; therapeutic use ; Animals ; Disease Progression ; Gene Expression ; Heterografts ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasms, Hormone-Dependent ; metabolism ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; genetics ; metabolism ; therapy ; Proto-Oncogene Proteins c-pim-1 ; metabolism ; Receptors, Androgen ; metabolism

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

OBJECTIVETo investigate the functional reconstruction technique for partial lower lip defects.

METHODS7 patients with lower lip cancer (3 cases of basal cell carcinoma, 2 cases squamous cell carcinoma and 2 cases papillary carcinoma) underwent excision. The full-thickness lower lip defects were one-third to two-third of the total lower lip length. All the defects were reconstructed with V-Y island advancement flaps based on the mental neurovascular bundle.

RESULTSThere was no flap loss. No recurrence was observed during the follow-up period of 3 months to one year. Both the aesthetic appearance, muscle function and sensation of the reconstructed lower lip were satisfactory.

CONCLUSIONSMental neurovascular V-Y island advancement flap is an ideal method for functional repair of partial lower lip defect.

Aged ; Carcinoma, Basal Cell ; surgery ; Chin ; innervation ; Female ; Humans ; Lip ; injuries ; Lip Neoplasms ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply

OBJECTIVETo describe the temporal trends in the mortality rate of gastric cancer during the period of 1988 and 2010, and to predict the gastric cancer mortality between 2016 - 2020.

METHODSThe data of gastric cancer mortality in Linzhou city between 1988 and 2010 was extracted from the cancer registry, including a total of 11 714 cases, covering 22 447 073 person-years. The mortality rate of gastric cancer of each 5-year period was calculated by sub-site and gender. Age-standardized rate (ASR) was calculated using the Chinese standard population in 1982. Intrinsic estimator (IE) model was used to fit the mortality trend by sub-site and gender, and to predict the mortality of gastric cancer in Linzhou city between 2016 and 2020.

RESULTSFrom 1988 to 2010, the gastric cancer mortality in Linzhou city was 52.18/100 000 (11 714/22 447 073) with the ASR at 49.23/100 000; the mortality in male was 67.02/100 000 (7678/11 455 512) with ASR at 68.68/100 000 while the mortality in female was 36.72/100 000 (4036/10 991 561) with ASR at 32.12/100 000. The mortality of cardia carcinoma was 27.87/100 000 (6257/22 447 073) with the ASR at 26.37/100 000; while the mortality of non-cardia carcinoma was 24.31/100 000 (5457/22 447 073) with the ASR at 22.86/100 000. The ASR of gastric cancer during 1988 - 1990 was 63.37/100 000 (1653 cases) and decreased by 28.34%, to 45.41/100 000 (2622 cases) during 2006 - 2010. The IE model showed that the birth cohort effect decreased greatly. The mortality risk of cardia carcinoma in population born after 1950s, decreased significantly; and the mortality risk of non-cardia carcinoma in population born in 20 century continually decreased. The death of gastric cancer among the population over 30 years old was predicted to be 3626 cases, increasing by 40.60% compared with the number between 2006 and 2010 (2579 cases). Among them, the mortality of cardia carcinoma increased by 51.89% (predicted number between 2016 and 2020 was 2456 cases, and 1617 cases between 2006 and 2010), and the mortality of non-cardia carcinoma increased by 21.62% (predicted number between 2016 and 2020 was 1170 cases, and 962 cases between 2006 and 2010).

CONCLUSIONThe mortality rate of gastric cancer in Linzhou city showed a decreasing trend during the period of 1988-2010, being mainly attributed to the cohort effect. However, the mortality will still increase in the future, between 2016 and 2020.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Forecasting ; Humans ; Male ; Middle Aged ; Registries ; Stomach Neoplasms ; epidemiology ; mortality

AIMTo prepare the liposomes which protect antisense oligodeoxynucleotides (ASON) against nuclease degradation and delivery ASON into cytoplasmic efficiently.

METHODSA cationic derivative of cholesterol, 3 beta-[N-(N',N'-dimethylaminoethan)-carbamoyl] cholesterol (DC-Chol) was synthesized and used to prepare cationic liposome. The characteristics of liposomes/ASON complexes including size, drug loaded efficiency and structure were investigated. Cellular uptake of fluorescence labled ASON (FAM-ASON) under different condition was determined by flow cytometric analysis. Denatured polyacryamide gel electrophoresis (DPGE) was used to analyze the role of liposomes in protecting ASON.

RESULTSThe mean values of preliposomes and liposomes/ASON complexes size were 185.7 and 228.2 nm, respectively. Cationic liposomes showed a high adsorption capacity for ASON. When the +/- charge ratio exceeded 2:1, more than 90% of the ASON was loaded into liposomes. Agarose gel electrophoresis showed three different existence of ASON in liposomes formulation: free, absorbed and encapsulated types. Concerning cellular uptake, DC-Chol liposomes indicated high efficient effect of increasing cellular uptake of ASON. Compared with free ASON, the total fluorescence intensity in cytoplasma was significantly enhanced. The level of increasing was largely depended on +/- charge ratio. The cellular uptake of FAM-ASON decreased in the presence of serum. The cellular total fluorescence intensity in 10% and 30% fetal bovine serum of cultured medium were only 22.3% and 15.5% as that of serum-free media, respectively. DPGE confirmed that free ASON was rapidly degraded by DNase I while ASON encapsulated into liposomes was efficiently protected.

CONCLUSIONThe cationic DC-Chol liposomes are shown to be promising carriers to deliver ASON into cytoplasma.

Cholesterol ; analogs & derivatives ; pharmacology ; Drug Carriers ; Drug Delivery Systems ; HeLa Cells ; Humans ; Liposomes ; pharmacology ; Multiple Myeloma ; pathology ; Oligonucleotides, Antisense ; administration & dosage ; blood ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo compare therapeutic effects of acupuncture and Chinese drug on oligospermia and asthenospermia in the male infertility.

METHODSTwo hundred and thirty-one cases were divided into 3 groups. The electroacupuncture group (n = 71) were treated with acupuncture at Qihai (CV 6), Guanyuan (CV 4), Zhongji (CV 3), etc., the Chinese drug group (n = 82) with oral administration of Chinese drug Wuzi Yanzong Pill and the acupuncture plus medication group (n = 78) with both electroacupuncture and oral administration of the Chinese drug. Changes of semen routine examination and the acrosome enzyme activity were observed before and after treatment in the 3 groups.

RESULTSThe effective rate was 67.6% in the electroacupuncture group, 68.3% in the Chinese drug group and 84.6% in the acupuncture plus medication group, the acupuncture plus medication group being significantly better than both the electroacupuncture and the Chinese drug group (P < 0.05); after treatment, the semen density, vitality and the acrosome enzyme activity were increased in the 3 groups, with more obvious increase in the acupuncture plus medication group.

CONCLUSIONBoth electroacupuncture and Chinese drug Wuzi Yanzong Pill can improve the semen quality, increase the pregnancy rate in the patient of male interfile with oligospermia and asthenospermia, and the effect of the combined treatment of acupuncture and Chinese drug is the best.

Acupuncture Therapy ; Adult ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Electroacupuncture ; Humans ; Infertility, Male ; drug therapy ; physiopathology ; therapy ; Male ; Sperm Count ; Sperm Motility