Objective To study local and systemic immune response in an animal model treated with recombinant hIFN-α-2b-BCG instillation. Methods The MB49 orthotopic bladder cancer model in C57BL/6 mice was established and treated separately with rBCG, wild BCG, wild BCG combined with IFN-α-2b and PBS as the control. The changes of lymphocyte subgroups in peripheral blood were analyzed with FCM, and mTNF-α and mIL-12 in peripheral blood of mice were detected with ELISA.Immunohistochemistry was carried out to detect the local immune reaction, T cell subsets and FAS, in bladder cancer after being treated with rBCG or wBCG. Results The content of CD4+ T lymphocyte was up-regulated in the rBCG group. The CD4+/CD8+ ratio of 2. 63 was up-regulated than pretreatment, significantly different than that of wBCG group(P＜0.05). ELISA assay showed that BCG significantly up-regulated the level of mTNF-α and mIL-12 in serum of orthotopie murine bladder cancer mice. The mTNF-α 806 pg/ml, mIL-12 860 pg/ml in rBCG group, was not significantly higher than those in wBCG group and combination group. The immunocompetent cell numbers with CD3, CD4,CD8 phenotype increased significantly in the tumor tissue of BCG treated group than the control(P＜0.05). The results of CD4+ in rBCG group and the combination group, and CD8+ in rBCG group were significantly higher than that of the wBCG(P＜0.05). The expression of Fas in tumor tissues treated with intravesical BCG was increased(P＜0. 05). Conclusions The recombinant IFN-α-2b-BCG can retrieve the disproportion of systemic lymphocyte subgroups, and increases Th1-type factors and local Fas expression in orthotopic murine bladder cancer. The recombinant IFN-α-2b-BCG is effective in regulating local and systemic immune reaction in orthotopic murine bladder cancer model.

OBJECTIVETo investigate the antitumor effect of recombinant IFN-alpha-2b-BCG on mouse bladder cancer MB49 cells in vitro, and to explore its antitumor mechanisms.

METHODSMB49 cells were co-cultured with recombinant BCG or wild BCG, and than were examined by light and transmission electron microscopy. The cell growth was assessed by MTT assay, and apoptosis rate and MHC-I of the MB49 cells was detected by flow cytometry using AO and Hoechst33258 fluorescence immunostaining.

RESULTSThe hIFN-alpha-2b-BCG-treated tumor cells showed slow growth, detachment of some cells, and various degree of degeneration. Light microscopy revealed organelle disorganization, chromatin aggregation, nuclear pyknosis, and cytolysis in some cells. Cellular membrane bulged and some bubbles were seen under fluorescence microscope using AO staining. Hoechst33258 assay also depicted frequent apoptosis in the tumor cells. The MTT assay showed that rBCG more actively than the wild BCG inhibited the proliferation of MB49 cells. The apoptosis rate of the recombinant BCG group was 19.7% and 46.6% at the time point of 24 h and 48 h, respectively, significantly higher than 10.8% and 20.9%, respectively, in the wild BCG group. The results of flow cytometry indicated that both types of BCG enhanced the expression of MHC-I in the MB49 cells, but more effective in the recombinant BCG group.

CONCLUSIONThe recombinant hIFN-alpha-2b-BCG has more strong immuno-modulatory properties, anti-tumor effect on MB49 cells and induces apparent cytotoxicity in the bladder cancer cells in vitro.

Adjuvants, Immunologic ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; BCG Vaccine ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytotoxicity, Immunologic ; Histocompatibility Antigens Class I ; metabolism ; Interferon-alpha ; pharmacology ; Mice ; Recombinant Proteins ; pharmacology ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo provide an appropriate angle and depth of needle insertion in acupuncture at Zusanli (ST 36) and avoid injuring the nerve and blood vessel and exert the most effect.

METHODSEighty adult lower-limb samples were used to dissect and observe the relative layered structures and adjoining important nerves and blood vessels in needling Zusanli (ST 36) according to the national standard.

RESULTSThe needling depth from the skin to the interosseous membrane and from the skin to posterior border of tibialis posterior is (2.22 +/- 0.31) cm and (4.42 +/- 0.53) cm, respectively. There are flabellate branches of anterior tibial arteries and deep peroneal nerves around the needle in the superficial layer of interosseous membrane. The vessel and nerve bundles containing tibial nerve and posterior tibial vessels can be touched when the needle body past through tibialis posterior.

CONCLUSIONIt is recommended that ideal average depth of acupuncture is 2.22 cm and the maximum depth is 4.42 cm for oblique needling Zusanli (ST 36). When it is injected, the needle should be perpendicularly inserted or deviated slightly to the direction of tibia and paralleled to medial surface of tibia. And the safe needling depth is generally less than 5 cm. The point of the body surface between tibialis anterior and extensor digitorum longus at 3 cun below Dubi (ST 35) is also an effectively stimulating point.

Acupuncture Points ; Female ; Humans ; Male ; Needles ; Tibia ; anatomy & histology

Objective According to the nursing characteristics of peacekeeping hospital of the United Nations in Lebanon, we applied the people oriented management and improved nursing quality.Methods With the guidance of people oriented management philosophy, we adopted various nursing management measures.Results The fulfillment rate of nursing work was 100%, and no nursing mistake or safety accident occurred. The satisfaction of patients was as high as 100%. All nursing staff obtained the peaceful merit badges awarded by Mr. Ban Ki-moon, the secretary-general of the United Nations.Conclusions Applying the people oriented management to nursing management effectively at peacekeeping hospital of the United Nations in Lebanon can promote the comprehensive quality of nurses and nursing quality.

OBJECTIVEThe purpose of this study is to explore the operation method and efficacy through retroperitoneal laparoscopy combined with urethral resection for treatment of renal pelvic carcinoma.

METHODSTotal nephroureterectomy with excision of bladder cuff by retroperitoneal laparoscopy plus urethral resection was performed in 18 patients with pathologically confirmed pelvic transitional cell carcinoma (II-III, T1N0M0-T2N0M0). The operation was performed using Olympus celioscope (30 degrees or 0 degree) under general anesthesia. First, a 10 mm incision was made at the intersection of midaxillary line and superior border 2 cm from crista iliaca, then a self-made hyponome filled with 250-300 ml water was put through the small incision in order to open the retroperitoneal space, followed by getting the hyponome out and perfusing CO2 into the retroperitoneal space to make a pneumoretroperitoneum. Finally, the celioscope was put into the retroperitoneal space to operate. During the operation, electric coagulation was used to stop bleeding and the bladder was not irrigated.

RESULTSThe operation was successfully performed in 18 patients without any complication. The operative time ranged from 150 to 190 min with a mean of 160 min. The hospital stay after operation was 7 to 10 days. There was no tumor recurrence or metastasis or implantation in all these patients after follow-up of 1-19 months.

CONCLUSIONCompared with regular operation mode, retroperitoneal laparoscopy plus urethral resection for treatment of renal pelvic carcinoma is a minimally invasive treatment with less bleeding and quick recovery.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Transitional Cell ; surgery ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; surgery ; Kidney Pelvis ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Nephrectomy ; methods ; Treatment Outcome ; Urethra ; surgery

OBJECTIVETo study the clinical outcomes of stage I testis teratoma, including pure teratoma, and to provide information on the treatment options for this disease.

METHODSWe retrospectively analyzed 27 cases of orchiectomy for stage I testis teratoma, excluding epidermoid cyst, and investigated its recurrence associated with treatment methods and clinicopathological factors.

RESULTSFour of the 27 cases relapsed, all in the orchiectomy group and confined to the retroperitoneal region, 3 with and the other 1 without risk factors, but with no death. No recurrence was found in those treated by orchiectomy followed by chemotherapy with bleomycin, etoposide and platinum (BEP). The total rate of recurrence was 15.8%. No severe side effects were observed in the 9 patients undergoing adjuvant BEP chemotherapy.

CONCLUSIONRisk factors may increase the recurrence rate of stage I testis teratoma, while postoperative adjuvant chemotherapy can reduce it, including that of pure teratoma, though surveillance policy remains the most popular option after orchiectomy.

Adolescent ; Adult ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Neoplasm Staging ; Retrospective Studies ; Teratoma ; pathology ; therapy ; Testicular Neoplasms ; pathology ; therapy ; Young Adult

BACKGROUNDThere are few reports of a biological role for glycosyltransferases in the infiltration of osteoarthritic synovitis. The aim of this research was to investigate the expression and cellular location of β-1,4-galactosyltransferase I (β-1,4-GalT-I) in a surgically-induced rat model of knee osteoarthritis (OA), and explore the role of β-1,4-GalT-I in the pathogenesis of OA.

METHODSMale Sprague-Dawley rats were randomly divided into three groups: OA group, sham group and normal group. The model of OA was established in the right knees of rats by anterior cruciate ligament transaction (ACLT) with partial medial meniscectomy. Fibroblast-like synoviocytes (FLSs) obtained from normal rat synovial tissue were cultured. The expression of β-1,4-GalT-I mRNA in the synovial tissue, articular cartilage and FLSs treated with tumor necrosis factor-α (TNF-α) were assayed by real-time PCR. Western-blotting and immunohistochemisty were used to observe the expression of β-1,4-GalT-I at the protein level. Double immunofluorescent staining was used to define the location of the β-1,4-GalT-I with macrophage-like synoviocytes, FLSs, neutrophils, and TNF-α in the OA synovium. The alteration of TNF-α in FLSs which were treated with lipopolysaccharide (LPS) and β-1,4-GalT-I-Ab were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe mRNA and protein expression of β-1,4-GalT-I increased in synovial tissue of the OA group compared with the normal and sham groups at two and four weeks after the surgery, however, no significant difference appeared in the articular cartilage. Immunohistochemistry also indicated that the β-1,4-GalT-I expression in OA synovium at four weeks after surgery increased sharply compared with the control group. β-1,4-GalT-I co-localized with macrophage-like synoviocytes, FLSs, neutrophils and TNF-α in rat OA synovitis. Moreover, in vitro β-1,4-GalT-I mRNA in FLSs was affected in a dose- and time-dependent manner in response to TNF-α stimulation. ELISA revealed that the expression of TNF-α was attenuated in FLSs in vitro when treated with anti β-1,4-GalT-I antibody.

CONCLUSIONβ-1,4-GalT-I may play an important role in the inflammation process of rat OA synovial tissue which would provide the foundation for further researching into the concrete mechanism of β-1,4-GalT-I in OA synovitis.

Animals ; Blotting, Western ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Galactosyltransferases ; genetics ; metabolism ; Immunohistochemistry ; Knee Joint ; enzymology ; pathology ; surgery ; Male ; Osteoarthritis, Knee ; enzymology ; genetics ; pathology ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; enzymology ; Synovitis ; enzymology ; etiology

AIMTo investigate the effect of androgen on the proliferation, differentiation and regression of canine prostatic stromal cells in vivo and human stromal cells in vitro.

METHODSTwenty-two dogs, including 15 normal prostate dogs and 7 prostatic hyperplasia dogs, had their serum concentration of testosterone and estrodiol determined by radioimmunoassay before and after castration. The expression of androgen receptor (AR) and estrogen receptor (ER) in the prostate were analysed by immunohistochemistry and semi-quantitative RT-PCR before and after castration. Light microscopy, transmission electron microscopy and TUNEL assay were carried out successively before and after castration to evaluate the prostatic histomorphology. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT). The proliferation of the cell culture was detected by MTT assay. The expression of TGFbgr, bFGF, AR, and smooth muscle cell (SMC) specific proteins (myosin and/or smoothelin) were detected using immunohistochemistry and RT-PCR. The differentiation from fibroblasts to smooth muscle cells was deduced by measuring the expression of SMC specific proteins.

RESULTSBefore castration, the serum concentrations of testosterone and estrodiol were not statistically different between normal and hyperplasia groups. Following castration, the serum concentration of testosterone decreased rapidly in 2 days, and the concentration of estrodiol had no significant change compared with the pre-castration data. In the prostate, AR was presented in both the epithelial and stromal cells and the AR mRNA level was higher in hyperplasia than in normal prostate tissues (P<0.05). While ER predominantly existed in the prostate stromal cells and the ER mRNA had no difference between the hyperplasia and the normal group. Within the early phase of castration (

CONCLUSIONThe whole prostate gland is an androgen-sensitive organ with both the epithelium and stroma under the control of androgen. Androgen may direct the proliferation, differentiation and regression of stromal cells by regulating the expression of TGFbgr, bFGF, AR and smooth muscle cell specific proteins.

Animals ; Biomarkers ; Cell Differentiation ; drug effects ; physiology ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Dihydrotestosterone ; pharmacology ; Dogs ; Estradiol ; blood ; Fibroblast Growth Factor 2 ; genetics ; pharmacology ; Gene Expression ; Humans ; Male ; Muscle, Smooth ; cytology ; physiology ; Orchiectomy ; Prostate ; cytology ; physiology ; Prostatic Hyperplasia ; physiopathology ; RNA, Messenger ; analysis ; Receptors, Androgen ; genetics ; Receptors, Estrogen ; genetics ; Stromal Cells ; cytology ; physiology ; Testosterone ; blood ; Transforming Growth Factor beta ; genetics ; pharmacology

Objective To construct recombinant human butyrylcholinesterase(rhBChE)knock-in goat fetal fibroblast cell lines(GFFs)by using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-mediated homology-directed repair mechanism for subsequent production of goat expressing rhBChE.Methods The efficient sgRNA sites targeting goat[3-casein(CSN2)gene were designed and screened,and the targeting efficiency of the sgRNA in goat mammary epithelial cells(GMECs)was confirmed by electro-transfection,flow sorting,and sequencing of PCR products.The red fluorescent reporter gene homology repair vector(P2A-mCherry)targeting the sgRNA was constructed,and then the integration and expression efficiency was detected by flow cytometry.The rhBChE homology repair vector(P2A-rhBChE)targeting the sgRNA of CSN2 gene was constructed in GFFs,the rhBChE positive cell clones were obtained via electro-transfection and flow sorting,and the rhBChE knock-in cell lines was identified by sequencing of PCR products.Results The sgRNA4 was identified as an efficient target of goat CSN2 gene,which could be also used for targeted integration of other genes.Three rhBChE knock-in cell lines were successfully constructed.Conclusion The rhBChE knock-in GFFs targeting goat CSN2 gene lays the foundation for the production of mammary bioreactors expressing rhBChE.

OBJECTIVETo study the basic therapeutic function of Tiaokou (ST 38).

METHODSAccording to clinically multi-central randomized controlled and single-blind test principle, 257 cases of periarthritis of shoulder were divided into two groups, a test group (n = 124) treated with oral anti-inflammatory analgesic medicine combined with acupuncture at Tiaokou (ST 38), and a control group (n = 133) treated with oral anti-inflammatory analgesic medicine. Their therapeutic effects were compared.

RESULTSThe total effective rate for stopping pain was 96.0% in the test group and 91.7% in the control group with a very significant difference between the two groups (P< 0.01). And the total effective rate for improvement of shoulder activity was 86.3% in the test group and 59.4% in the control group with a very significant difference between the two groups (P<0.01).

CONCLUSIONOral anti-inflammatory analgesic medicine combined with acupuncture has obvious therapeutic effect on periarthritis of shoulder, which is better than that of simple oral anti-inflammatory analgesic medicine.

Humans ; Periarthritis ; therapy ; Shoulder ; Shoulder Pain ; therapy ; Single-Blind Method