OBJECTIVETo establish the method of promoting human peripheral blood mononuclear cell proliferation by polysaccharide of snakegourd root and identify the effects of polysaccharide of snakegourd root on lymphocyte proliferation, T lymphocyte subsets and the different levels of TNF-alpha and IL-6.

METHODThe polysaccharide of snakegourd root preparations were purified with dialysis and ethanol precipitation. The healthy human PBMC were used as the target cells for screening potency of the drugs. MTT colorimetry was established to examine the levels of lymphocyte proliferation on human PBMC by polysaccharide of snakegourd root in vitro. The percents of lymphocyte subsets (CD3+, CD4+, CD8+ T lymphocyte) and the different levels of TNF-a and IL-6 in PBMC were analysed by FCM and ELISA, respectively.

RESULT1.0-50.0 mmol x L(-1) of polysaccharides of snakegourd root showed the significant effects of promoting proliferation of human PBMC (P < 0.05). The percents of CD3+, CD4+, CD8+ T lymphocytes in PBMC treated with 5.0 and 10.0 mmol x L(-1) of polysaccharides of snakegourd root were significantly higher than those of the control group (P < 0.05). The levels of TNF-alpha and IL-6 were significantly higher than those of the control group after 1.0, 5.0, 10.0 mmol x L(-1) of polysaccharides of snakegourd root stimulation on the human PBMC at 8 hours (P < 0.05).

CONCLUSIONThe significant effects on promoting lymphocyte proliferation and activation of the polysaccharide of snakegourd root are confirmed in this study. The percents of lymphocyte subsets are increased in different degrees by the polysaccharide of snakegourd root. The high levels of TNF-alpha and IL-6 are secreted after the polysaccharides of snakegourd root stimulation on the human PBMC, which lays a foundation for further elucidating the immunocompetence effects and mechanism of the polysaccharide of snakegourd root.

Dietary Carbohydrates ; Humans ; Immunocompetence ; drug effects ; Polysaccharides

Fungi immunoregulatory proteins family is effective in immunological regulation and anti-tumor. We used Pichia pastoris expression system for recombinant expression of Lz-8, an immunomodulatory protein isolated from fruiting body of Ganoderma lucidum. The Gs115 (mut+) strains of P. pastoris was used as host cells. PCR and sequencing of DNA showed that Lz-8 cDNA was successfully integrated into the P. pastoris genome. Electrophoresis (SDS-PAGE), matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) and immunological techniques were used to identify recombinant Lz-8 (rLz-8). Lz-8 expressed in Escherichia coli, the Pichia system requires further optimization to obtain more active fungi immunomodulatory protein. Lz-8 was expressed in P. pastoris successfully, and polyacrylamide gel electrophoresis in the presence of SDS-PAGE gave a single band with an apparent Mr=14,000 D. MALDI-TOF-MS also showed that molecular weight of rLz-8 was 12,722 D. Aggregation was observed from sheep red blood cells in the presence of purified rLz-8 within the concentration range of 12.5-50 microg/mL. However, no aggregation was seen at concentration greater than 50 microg/mL for any type of human red blood cell. The dose at 0.5 mg/kg of rLz-8 induced macrophage cytophagocytesis, and set interferon as control at 0.5 mg/kg. These results suggested that active and stable rLz-8 was obtained in P. pastoris expression system.
Fungal Proteins ; biosynthesis ; genetics ; immunology ; Immunocompetence ; immunology ; Macrophages ; immunology ; Phagocytosis ; drug effects ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

This study was aimed to investigate the cytotoxic effect of the Naja Naja Actra Venom Component (NNAVC) combined with activated immune cells on human acute myeloblastic leukemia line KG1a cells. The cytotoxic effects of NNAVC at different concentrations on KG1a cells were measured by CCK-8 method. LDH releasing assay was used to detect the cytotoxic effects of activated immune cells, NNAVC combined with activated immune cells on KG1a cells and the sensitivity of KG1a treated with NNAVC to activated immune cells. The results showed that the inhibitory rate of NNAVC on KG1a cells increased with the concentration enhancement, the cytotoxicity of activated immune cells at the different effector to target (E:T) ratios(6.25:1, 12.5:1, 25:1) on KG1a cells were 12.30%, 24.85% and 52.26%. The cytotoxicity of NNAVC combined with activated immune cells at the different E:T cell ratios (10:1, 20: 1) on KG1a cells were 56.21% and 85.59%, which were higher than that of NNAVC or activated immune cells alone. The cytotoxicity of activated immune cells at the E: T cell ratio of 10:1 on KG1a cells treated with NNAVC at different concentrations were 25.65%, 31.33%, 28.63% and 16.78%, respectively, and that at the E:T cell ratio of 20: 1 were 40.62%, 44.70%, 44.62% and 40.72%. It is concluded that:both of NNAVC and activated immune cells have lethal effect on KG1a cells, and the combination of NNAVC and activated immune cells can strengthen their effect on KG1a.
Animals ; Cell Line, Tumor ; drug effects ; Cytotoxicity, Immunologic ; Elapidae ; Humans ; Immunocompetence ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Venoms ; pharmacology

PURPOSE: Bone metastasis invariably increases morbidity and mortality. This study compares the effects of ibandronate and paclitaxel on bone structure and its mechanical properties and biochemical turnover in resorption markers using an immunocompetent Walker 256-Sprague-Dawley model, which was subjected to tumor-induced osteolysis. MATERIALS AND METHODS: Seventy rats were divided equally into 4 groups: 1) sham group (SHAM), 2) tumor group (CANC), 3) ibandronate treated group (IBAN), and 4) paclitaxel treated group (PAC). Morphological indices [bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp)] and mechanical properties (failure load, stiffness) were evaluated after thirty days of treatment period. Bone resorption rate was analysed using serum deoxypyridinoline (Dpd) concentrations. RESULTS: Morphological indices showed that ibandronate (anti-resorptive drug) had a better effect in treating tumor-induced architectural changes in bone than paclitaxel (chemotherapeutic drug). The deterioration in bone architecture was reflected in the biomechanical properties of bone as studied with decreased failure load (F(x)) and stiffness (S) of the bone on the 30th day postsurgery. Dpd concentrations were significantly lower in the IBAN group, indicating successful inhibition of bone resorption and destruction. CONCLUSION: Ibandronate was found to be as effective as higher doses of paclitaxel in maintaining stiffness of bone. Paclitaxel treatment did not appear to inhibit osteoclast resorption, which is contrary to earlier in-vitro literature. Emphasis should be placed on the use of immunocompetent models for examining drug efficacy since it adequately reflects bone metastasis in clinical scenarios.
Amino Acids ; Animals ; Biomechanical Phenomena/*drug effects/physiology ; Bone Density/drug effects/physiology ; Bone Neoplasms/*drug therapy ; Bone Resorption/*chemically induced ; Diphosphonates/*pharmacology ; Immunocompetence ; Male ; *Neoplasm Metastasis ; *Osteolysis ; Paclitaxel/*pharmacology ; Rats ; Rats, Sprague-Dawley

We report a case of cytomegalovirus (CMV) retinitis after intravitreal bevacizumab injection. A 61-year-old woman with diabetic macular edema developed dense vitritis and necrotizing retinitis 3 weeks after intravitreal bevacizumab injection. A diagnostic vitrectomy was performed. The undiluted vitreous sample acquired by vitrectomy was analyzed by polymerase chain reaction and culture. Polymerase chain reaction of the vitreous was positive for CMV DNA. Other laboratory results did not show evidence of other infectious retinitis and systemic immune dysfunction. Human immunodeficiency virus antibodies were also negative. After systemic administration of ganciclovir, retinitis has resolved and there has been no recurrence of retinitis during the follow-up period of 12 months. Ophthalmologists should be aware of potential risk for CMV retinitis after intravitreal bevacizumab injection.
Angiogenesis Inhibitors/administration & dosage/adverse effects ; Antibodies, Monoclonal, Humanized/administration & dosage/*adverse effects ; Cytomegalovirus/genetics ; Cytomegalovirus Retinitis/diagnosis/*etiology/immunology ; DNA, Viral/analysis ; Diagnosis, Differential ; Female ; Humans ; Immunocompetence/*drug effects ; Intravitreal Injections ; Macular Edema/diagnosis/*drug therapy ; Middle Aged ; Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A/antagonists & inhibitors

OBJECTIVETo investigate the efficacy and mechanism of Bushen Huoxue Recipe (, BHR) in the treatment of murine autoimmune premature ovarian failure (POF).

METHODSThe recombinant porcine zona pellucida 4 (pZP4) was expressed in E. coli BL21 (DE3) strain within prokaryotic plasmid pET28a (+), purified by Ni-affinity chromatography and verified by Western blot. Murine autoimmune POF model was established by immunization with pZP4 of female BALB/c mice. Fifty POF mice were randomly divided into 5 groups, which were respectively given low (3.75 mg/kg), moderate (7.5 mg/kg), and high dose (15.0 mg/kg) of BHR by gastrogavage once daily for 20 days, with 17-β-estradiol (0.13 mg/kg) and normal saline as positive and negative control. Estrous cycles were analyzed through vaginal smears, serum estradiol (E) levels, and anti-pZP4 antibody titers were detected by ELISA. The proliferative responses in vitro of spleen lymphocytes to pZP4 antigen restimulation were measured by [(3)H]-thymidine incorporation, and the histomorphology changes of ovary were evaluated by optical microscope.

RESULTSThe purified pZP4 was visible as a single lane with 14.4 kD in SDS-PAGE and Western blot. The murine POF model with lengthening estrous cycles, decreased levels of serum E2, high titers of serum anti-pZP4 antibody, and reduced ovarian follicles and corpus lutea were established by immunization with recombinant pZP4. Treatment with moderate and high dosage BHR significantly increased ovarian follicles and reduced the proliferation of spleen lymphocytes to the pZP4 antigen of POF mice (P <0.05). However, only the high dosage BHR administration significantly improved the estrous cycles, elevated the serum E levels (P <0.01), and decreased the serum anti-pZP4 antibody titers of model mice P<0.05).

CONCLUSIONSThe recombinant pZP4 could evoke the antigen-specific immune response in mice and induce the autoimmune ovarian injury. It has been demonstrated that BHR was able to increase the serum E levels and protect ovarian functions from the autoimmune injury in murine POF model.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Egg Proteins ; immunology ; isolation & purification ; Female ; Immunization ; Immunocompetence ; drug effects ; Membrane Glycoproteins ; immunology ; isolation & purification ; Mice ; Mice, Inbred BALB C ; Ovary ; drug effects ; immunology ; pathology ; Primary Ovarian Insufficiency ; drug therapy ; immunology ; pathology ; Receptors, Cell Surface ; immunology ; isolation & purification ; Recombinant Proteins ; immunology ; isolation & purification ; Sus scrofa ; Zona Pellucida Glycoproteins

Burkholderia cepacia (B. cepacia) infection is rarely reported in an immunocompetent host. It is a well known occurence in patients with cystic fibrosis and chronic granulomatous disease where it increases both morbidity and mortality. It has also been included in the list of organisms causing nosocomial infections in an immunocompetent host, most of them transmitted from the immunocompromised patient in which this organism harbors. We report a rare case of isolation of B. cepacia from the bronchoalveolar lavage fluid of an immunocompetent agriculturist who presented with productive cough and fever associated with a pyopneumothorax. This is the first case of community acquired infection reported in an immunocompetent person in India.
Adult ; Anti-Bacterial Agents ; therapeutic use ; Azithromycin ; therapeutic use ; Bronchoalveolar Lavage Fluid ; microbiology ; Burkholderia Infections ; diagnosis ; drug therapy ; transmission ; Burkholderia cepacia ; drug effects ; isolation & purification ; Ceftazidime ; therapeutic use ; Ceftizoxime ; therapeutic use ; Community-Acquired Infections ; diagnosis ; drug therapy ; Drug Resistance, Multiple, Bacterial ; Humans ; Immunocompetence ; India ; Male ; Pneumothorax ; complications