Two lipase active fractions Lip1 and Lip2 were purified from the cell extract of Rhizopus chinensis CCTCCM201021.Both gave a single band on SDS-PAGE after using ammonium sulfate precipitation、Phenyl-Sepharose FF、DEAE-Sepharose FF and Sephadex G100 gel filtration chromatographies.The molecular masses of two lipases were 59.2kD and 39.4kD respectively.Lip1 and Lip2 showed optimal pH at 8.0 and 8.5 and their optimal temperatures were 40℃ and 35℃ respectively.The substrate specificity of the two lipases was obviously different.Lip1 was more specific to long chain fatty acid of p-nitrophenyl esters while Lip2 had a preference for the hydrolysis of short chain fatty acid of p-nitrophenyl esters.Lip1 had 1,3-position specificity for triacylglycerols hydrolysis while Lip2 had nonspecific position.Both lipases were stimulated by Ca 2+、Mg 2+ while SDS had strong inhibition on their activities.Lip1 and Lip2 had good stability in cyclohexane、hexane、heptane and isooctane(30% V/V).

To evaluate the clinical efficacy and safety of Qinghouyan lozenge in the treatment of acute pharyngitis due to Lung-heat and Yin-deficiency, and compare with Qinghouyan oral Liquid. Totally 144 subjects were enrolled and randomly divided into two groups (72 in the test group and 72 in the control group). The participants in the test group were given Qinghouyan lozenge for 5 days, and those in the control group were given Qinghouyan oral Liquid for 5 days. The effectiveness evaluation indexes were pharyngalgia/odynophagia disappearance rate, overall efficacy of TCM syndromes, TCM syndrome scores, and single syndrome and sign disappearance rate. During the test, the safety was evaluated by vital sign, lab examination indexes and adverse events. The results for the full analysis set showed that the couth disappearance rate, the incidence rate of TCM syndromes, and the throat/uvula congestion disappearance rate of the test group were higher than that of the control group (P < 0.05), with significant differences in the changes in syndrome scores between the two groups (P < 0.05). Altogether 3 adverse events were observed in the test group while 6 adverse events in the control group, without significant differences in the adverse event rate between the two groups (P < 0.05), serious abnormal laboratory examinations and vital signs. In conclusion, Qinghouyan lozenge has better efficacy in treatment of acute pharyngitis due to Lung-heat and Yin-deficiency than Qinghouyan oral liquid, with good safety.
Acute Disease ; Double-Blind Method ; Humans ; Medicine, Chinese Traditional ; Pharyngitis ; drug therapy

Objective To investigate the clinical outcomes of patients undergoing microsurgery for cerebellar medulloblastoma. Methods This retrospective analysis of the clinical and follow-up data involves 57 patients who received microsurgery for pathologically confirmed cerebellar medulloblastorna, and the microsurgical techniques for medulloblastoma were discussed. Results Among the 57 patients, 42 had total tumor resection, 13 had subtotal and 2 had partial resection of the tumors. Patency of the midbrain aqueduct was achieved in all the cases after the surgery. Hydrocephalus was found in 43 patients before the operation and only in 16 patients after the operation. Tumor relapse occurred in 19 patients 2 years after the operation, including 8 with implantation metastasis compromising the central nervous system and 1 patient with frontal lobe metastasis who received reoperations. The earliest tumor relapse occurred 20 days after the surgery. The 2- and 5-year postoperative survival rates in these patients were 68.4% and 49.1%, respectively. Conclusion Good therapeutic effects can be achieved with total resection of the tumors and postoperative whole brain and spinal cord radiotherapy in patients with medulloblastomas.

BACKGROUNDMany epidemiological and experimental evidences prove that cervical cancers are strongly associated with genital high-risk types of human papillomavirus (HPV). HPV16 is present in 50% of the tumor specimens. Thus, it is important to develop vaccines against HPV16 and cervical cancer. The authors studied the expression of the HPV16 L1DeltaCE7N fusion protein in E. coli and observed its immunogenicity.

METHODSThe fragment of HPV16 L1DeltaC gene and the E7N gene were amplified by PCR separately; the fusion gene named L1DeltaCE7N was generated by fusing E7N to the C terminal of L1DeltaC then the chimeric gene was cloned into prokaryotic expression vector pGEX-2T and expressed in E. coli strain JM109. The L1DeltaCE7N protein expressed were detected by Western blot. Finally its immunogenicity was characterized in immunized mice.

RESULTSIt was proved that the sequence and open reading frame of fusion gene L1DeltaE7N was correct by sequencing; SDA-PAGE gel analysis showed that HPV16 L1/E7 fusion protein was highly expressed in E. coli; the protein was expressed as soluble form and the molecular weight was about 85 x 10(3). The fusion protein could be purified by affinity chromatography and gel filtration. The ELISA result indicated that L1/E7 could elicit specific antibodies against L1 and E7 in immunized mice. In vivo tumor protection test indicated that tumor formation was retarded or prevented in the mice after vaccination with L1/E7, when C57 BL/6 mice were challenged by syngeneic HVP16E6 and E7 transformed tumor cells.

CONCLUSIONHPV16L1/E7 fusion protein was expressed in E. coli, it can be a candidate for prophylactic and therapeutic vaccine for HPV16-associated infection and tumors.

Animals ; Blotting, Western ; Cell Line, Tumor ; Escherichia coli ; genetics ; Female ; Humans ; Immunization ; methods ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplasms, Experimental ; immunology ; pathology ; prevention & control ; Oncogene Proteins, Fusion ; genetics ; immunology ; metabolism ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Papillomaviridae ; genetics ; immunology ; metabolism ; Papillomavirus Infections ; immunology ; pathology ; prevention & control ; Papillomavirus Vaccines ; administration & dosage ; immunology ; Recombinant Fusion Proteins ; immunology ; metabolism ; ultrastructure

OBJECTIVETo evaluate clinical outcomes after laparoscopic total mesorectal excision (TME) combined with intersphincteric resection (ISR) for ultra-low rectal tumors.

METHODSClinical data of 36 patients with ultra-low rectal tumor undergoing laparoscopic TME combined with ISR were analyzed retrospectively.

RESULTSThe median distance from the inferior margin of the tumor to the anal verge was 3.4 (2.0-5.0) cm. There were 33 cases of well/moderately differentiated adenocarcinoma and 3 rectal malignant villous adenoma. There were 16 patients with stage I disease, 15 with stage II A, 3 with stage III A, and 1 with III B. Postoperatively, one patient developed stenosis at the end ileostomy and 3 anastomotic leakage. After a median follow-up of 16(4-49) months, one patient developed local recurrence at the anastomosis and one case died of liver metastasis. In the 19 patients who had a minimum follow-up of one year, the bowel movements frequency ranged from 1-4 times per day, and these patients were able to withhold defecation for more than 5 minutes.

CONCLUSIONSLaparoscopic TME combined with ISR can achieve oncologic clearance, sphincter preservation, and minimal invasiveness for ultra-lower rectal cancer. However, patients selection should be cautious.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Mesentery ; surgery ; Middle Aged ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Retrospective Studies ; Treatment Outcome

Actins ; metabolism ; Animals ; Brain ; cytology ; metabolism ; Cell Culture Techniques ; methods ; Cells, Cultured ; Immunohistochemistry ; Pericytes ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; Staining and Labeling

OBJECTIVETo investigate the anti-tumor immunity of the non-replicating recombinant vaccinia virus expressing HPV16 E6 and E7 proteins.

METHODSC57BL/6 mice were immunized by non-replicating recombinant vaccinia virus (NTVJmE6E7), and then specific CTLs were determined. Immune protection effects were evaluated by challenges of different doses of TC-1 tumor cells. Immunotherapeutic effects in form of recurrence were evaluated on the tumor-removed mice.

RESULTSMice immunized by NTVJmE6E7 could generate TC-1 cell specific cytotoxic T lymphocyte (CTL). Mice boosted with NTVJmE6E7 could tolerate the challenge of 1 x 10(4) TC-1 cells. NTVJmE6E7 could effectively prevent the tumor recurrence in the tumor-removed mice.

CONCLUSIONNTVJmE6E7 can be taken as a candidate of therapeutic vaccine for HPV-associated tumors and their precursor lesions.

Animals ; Cancer Vaccines ; Cells, Cultured ; Female ; Genetic Vectors ; Immunotherapy ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplasms, Experimental ; immunology ; therapy ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccinia virus ; genetics ; Viral Vaccines

OBJECTIVETo study the incidence and risk factors of HIV and syphilis seroconversion among men who have sex with men (MSM) in Beijing.

METHODSA total of 550 MSM were recruited on the basis of community and followed up after 6 and 12 months in Beijing. Each subject was investigated by only one investigator at one time to collect information on demographics and behaviors. Blood samples were collected to test HIV and syphilis seroconversion. ELISA was used for screening test, west blotting (WB) and Particle agglutination were used for confirmatory test.

RESULTSA total of 550 MSM investigated, among which 4.5% (25/550) were HIV-positive and 29.3% (161/550) were syphilis-positive. For 525 HIV-negative MSM, 87.0% (457/525) retained during the 12-month investigation. Seroincidence for HIV and syphilis were 3.37/100 person-years (95%CI = 1.66 - 5.08) and 9.32/100 person-years (95%CI = 5.87 - 12.77) respectively. HIV seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 7.11/100 and 0.76/100 person-years respectively. Multivariate Cox regression analysis revealed that rectal douching after homosexual anal intercourse in the past 3 months (HR = 9.23, 95%CI = 2.08 - 40.88) was significantly associated with HIV seroconversion. Syphilis seroconversions for those who met male sex partners in parks, public washrooms or bathhouses in the past 3 months were 41.77/100 and 7.97/100 person-years respectively. Syphilis seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 16.17/100 and 4.92/100 person-years respectively. In the past 3 months, meeting male sex partners in parks, public washrooms or bathhouses (HR = 4.67, 95%CI = 1.77 - 12.34) and performing rectal douching after homosexual anal intercourse (HR = 3.09, 95%CI = 1.40 - 6.83) were significantly associated with syphilis seroconversion.

CONCLUSIONThe seroconversions of HIV and syphilis during the follow-up visits in this MSM cohort study in Beijing were very serious, and that the associated factors for seroconversions were rectal douching after homosexual anal intercourse and meeting male sex partners in parks, public washrooms or bathhouses.

Adolescent ; Adult ; Antibodies, Bacterial ; blood ; China ; epidemiology ; HIV ; immunology ; HIV Antibodies ; blood ; HIV Infections ; blood ; epidemiology ; HIV Seropositivity ; blood ; epidemiology ; Homosexuality, Male ; Humans ; Incidence ; Male ; Risk Factors ; Sexual Behavior ; Syphilis ; blood ; epidemiology ; Treponema pallidum ; immunology ; Young Adult

OBJECTIVETo generate a human papillomavirus (HPV16) prophylactic and therapeutic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identified.

RESULTSDNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaccinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were co-expressed in CEF cells infected with the recombinant virus.

CONCLUSIONNTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations.

Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cells, Cultured ; Chick Embryo ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomaviridae ; genetics ; immunology ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; immunology ; prevention & control ; virology ; Papillomavirus Vaccines ; genetics ; immunology ; therapeutic use ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Transfection ; Tumor Virus Infections ; immunology ; prevention & control ; virology ; Uterine Cervical Neoplasms ; immunology ; prevention & control ; virology ; Vaccinia virus ; genetics ; Virus Replication

Objective:To investigate the effect of vasoactive intestinal peptide on IL-1β expression after corneal alkali burn.Methods:27 healthy adult female Wistar rats were randomly divided into group A(normal blank control group),group B(alkali burn treated with saline group),and group C (alkali burn treated with vasoactive intestinal peptide group).Group A were not processed.Group B,C were made right eyes′moderate corneal alkali burn models.After inducing burn,group B and group C were followed respectively by 0.9% saline and 0.25 g/L vasoactive intestinal peptide eye drops,4 times per day.The corneal epithelial healing rates were calculated by measuring the corneal staining area by methylene blue staining at 3 d,7 d and 14 d after inducing burn.At 3 d,7 d and 21 d,the corneal tissues were removed and divided into two equal parts.The corneal tissues were observed by staining,and the expression of IL-1β were detected by immunohistochemistry.Results:The healing rates of corneal epithelium in group C were greater than those in group B at 7 d and 14 d(P<0.05).The mean optical density of IL-1β was measured by immunohistochem-istry.The mean optical density in group B and group C at 3 d were(1.62 ± 0.96) ×10-2,(0.98 ± 0.45) ×10-2,respectively,which at 7 d were (1.42 ± 0.62) ×10-2,(0.71 ± 0.22) ×10-2,respectively,which at 21 d were(0.37 ± 0.24) ×10-2,(0.13 ± 0.16) × 10-2,respectively.The mean optical density of corneal IL-1β in group C were lower than those in group B(P<0.05).Conclusion:Va-soactive intestinal peptide can decrease the expression of IL-1β after corneal alkali burn,down-regulate the level of immune stress, reduce the inflammatory reaction and promote the recovery of cornea after alkali burn.