Objective To detect the neurofibromin expression and observe the biological features of the osteoblasts of patients with type 1 neurofibromatosis and scoliosis. Methods 10 cases of congenital scoliosis and 8 cases of NF1 scoliosis were chosen. The two groups were with the similar age(with the average of 11.7 years and 12.5 years) and Cobb angle(with the average of 85? and 94?). Cancellous bone was harvested from the ilia and the bone explants culture system was used. Proliferation of the osteoblasts, and also the specific differentiation index including alkaline phosphatase, type Ⅰ collagen and osteocalcin was assayed in sewnd generation osteoblasts. Neurofibromin expression in the two kinds of osteoblasts was detected with immunoprecipitation followed by Western blot. Results Compared to the osteoblasts of patients with congenital scoliosis, lower level of neurofibromin was expressed in osteoblasts of patients with NF1 scoliosis, (the OD value was 1.05?0.06 and 2.59?1.40 respectively, P=0.002). The level of alkaline phosphatase, type Ⅰ collagen and osteocalcin were significant lower in osteoblasts of patients with NF1 scoliosis than those of CS patients (44.69 IU/mg vs 51.38 IU/mg, P=0.019; 226.34 ng/mg vs 249.93 ng/mg, P=0.014; 7.41 ng/mg vs 8.87 ng/mg, P=0.049). But the proliferation rate of the NF1 osteoblasts was significant higher than that of the CS osteoblasts (3.34 and 2.70 respectively, P=0.049). Conclusion With the decrease of neurofibromin expression, the NF1 osteoblasts show some defects in their function, these function defects may contribute to varieties of skeletal abnormalities as dystrophic change and decreased bone density.

Abdominal Neoplasms ; diagnosis ; diagnostic imaging ; Adolescent ; Autoantibodies ; blood ; Diagnosis, Differential ; Female ; Humans ; Mouth Mucosa ; pathology ; Paraneoplastic Syndromes ; diagnosis ; immunology ; pathology ; Pemphigus ; diagnosis ; immunology ; pathology ; Skin ; pathology ; Tomography, X-Ray Computed

In order to reaction the quality present situation, problems on the current quality of animal sources of drugs are summed up by using test data analysis, literature search and marketing research. This paper can also help the improvement of the quality management, the revision of the relevant department policy system and the improvement of standards.
Animals ; China ; Medicine, Chinese Traditional ; standards ; Pharmaceutical Preparations ; analysis ; standards ; Quality Control

HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.

Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.

The pol gene of HIV-1 encodes mainly three enzymes: reverse transcriptase (RT), protease (PR) and integrase (IN). Currently, FDA approved drugs targeting RT and PR are available and administered in various combinations, while no anti-IN drug was approved. HIV-1 integrase is an essential enzyme for the viral replication and an interesting target for the design of new pharmaceuticals for multi-drug therapy of AIDS. The 288 amino acids of IN (32kDa) recognizes specific sequences in the long terminal repeats (LTRs) of the retroviral DNA. The IN protein catalyzes the 3′-processing step and the 5′-strand transfer step reaction in vivo, which was called integration and this reaction could be analysed by ELISA Assay in vitro. It has been reported that F185K and C280S mutations of HIV-1 integrase would improve the enzyme solubility, and the catalytical activity of the enzyme was the same as that of the wild-type enzyme in vitro. In order to build the platform of screening inhibitor against integrase of HIV-1 virus, the IN enzyme was expressed and the function of integrase protein was assayed. The cDNA of clade B HIV-1 genome was used as a template, overlapped PCR was used to construct site mutagenesis of F185K/C280S and NdeI/Xho I restriction sites were brought in. The PCR product was cloned into the prokaryotic vector pET-28a(+) to form a recombined plasmid, transferred into the host cell E.coli(BL21 DE3). The recombined clones were identified by PCR and Nde I/Xho I digestion .The positive plasmid was sequenced, and the successfully recombined plasmid in the host cell was induced by IPTG. The expressed IN protein was puriied sy the Co+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombined IN protein. The recombinant protein was soluble, and expressed highly and stably in E.coli. The molecular weight of the expression product was identical to the expectation. The IN protein was proved to be functional in 3′ processing and 5′strand transfer by ELISA. It will be helpful to build the platform of screening inhibitors against HIV-1 integrase.

Objective To discuss the clinical characteristics and treatments for chondrosarcoma of paranasal sinus and the skull base. Methods The clinical characteristics of chondrosarcoma of paranasal sinus and skull base in 7 patients tmderwent endoscopic surgeries between 2001 and 2008 were analyzed. Of the patients,4 men and 3 women. The patients' age ranged from 18 to 47 years,with a median of 31 years. Clinical symptoms: stuffy, nose bleeding, runny, headache, diplopia, eye outreach limited, blurred vision and even blindness. Surgery methods:under nasal endoscopy,after the attachment sites of the tumors to normal tissues were confirmed,the tumors were peeled off along the clear boundary between the tumors and normal tissues,and the potential residual tumor tissues on bones were cleared by a drill. Results The patients were followed up postoperatively for 24 to 108 months,with a median of 36 months. Five of 7 patients were no recurrence,2 were alive with tumor. Conclusions Chondrosarcoma of paranasal sinus and skull base can be treated by nasal endoscopic surgery, with good clinical outcome.

OBJECTIVETo explore the expressions of HO-2 and CO in the corpus cavernosum of castrated rats in order to further study the pathogenesis of erectile dysfunction (ED).

METHODSWe randomly divided 72 male SD rats into four groups: normal control, sham operation, castration, and castration + ZnPP. We detected intracavernous pressure (ICP) and penile erection in the basic condition and after apomorphine (APO) induction, determined the expression of the HO-2 protein in the corpus cavernosum by laser scanning confocal microscopy, and measured the level of CO by spectrophotometry during different periods of penile erection.

RESULTSThe ICP in the basic condition and that after APO induction and the rate of penile erection were decreased significantly in the castration group ([11.68 ± 0.69] mmHg, [54.81 ± 3.86] mmHg, and 33.3%) and the castration + ZnPP group ([11.20 ± 0.71] mmHg, [41.17 ± 5.41] mmHg, and 22.2%) as compared with the normal control ([22.83 ± 2.66] mmHg, [66.92 ± 7.77] mm-Hg, and 100%) and the sham operation group ([23.35 ±2.22] mmHg, [70.43 ?7. 22] mmHg, and 100%) (all P <0. 01). After APO induction, ICP in the castration + ZnPP group was remarkably reduced in comparison with that in the castration group (P < 0.01), and so was the expression of the HO-2 protein before and during penile erection in the castration (445.4 ± 23.7 and 847.4 ± 35.0) and the castration + ZnPP group (390.1 ± 29.7 and 526.0 ± 52.5) compared with the normal control (512.7 ±57.4 and 1145.2 ± 89.8) and the sham operation group (583.7 ± 8.0 and 1016.3 ± 79.8), the expression of the HO-2 protein significantly decreased in the castration group (445.4 ± 23.7 and 847.4 ± 35.0) (P < 0.05 or 0.01), markedly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01) but with no significant differences among the four groups after it. Before, during and after penile erection, the levels of CO were remarkably decreased in the castration ([20.59 ± 1.01], [32.53 ± 1.26], and [18.71 ± 1.22] x 10(-7) nmol/L) and the castration +ZnPP group ([12.52 ± 1.05], [21.90 ± 1.02], and [16.56 ± 0.55] x 10(-7) nmol/L) as compared with the normal control ([26.76 ± 1.41], [48.25 ± 1.01], and [27.10 ± 1.58 ] x 10(-7) nmol/L) and the sham operation group ([25.41 ± 2.09], [ 47.90 ± 1.22], and [25.67 ± 1.20] x 10(-7) nmol/L) (P < 0.05 or 0.01), significantly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01).

CONCLUSIONDecreased expressions of HO-2 and CO may correlate with erectile dysfunction in castrated rats.

Animals ; Apomorphine ; pharmacology ; Carbon Monoxide ; metabolism ; Dopamine Agonists ; pharmacology ; Erectile Dysfunction ; etiology ; Humans ; Male ; Molecular Chaperones ; metabolism ; Orchiectomy ; Penile Erection ; drug effects ; Penis ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of Qingre Liyan Decoction (QRLYD) in the prevention and treatment of acute radiative oral mucositis (AROM), and to explore the mechanism of QRLYD by detecting epidermal growth factor (EGF) and T lymphocytes (CD3, CD4, and CD8).

METHODSSixty patients conforming with the standard were randomly assigned to two groups, 30 patients in each group. Patients in the trial group were treated with QRLYD, and those in the control group were treated with Dobell's solution, both groups receiving conventional radiation treatment. The treatment course for both groups was 6 weeks on average. Blood routine test, CD3, CD4, and CD8 in the peripheral blood and EGF in the saliva were detected one day before and on the 14th and 28th day of radio-therapy.

RESULTSPatients in the trial group were in good condition with normal spirits and intake of food and drinks. The incidence of AROM is lower and the effect in preventing AROM is higher in the trial group than those in the control group (P<0.05). The EGF in saliva, and CD4 and CD8 in the blood of patients in the trial group were higher than those in the control group (P<0.05).

CONCLUSIONQRLYD can cure and prevent AROM. The mechanism may be related with its effects in enhancing body immunity and promoting salivary EGF.

Acute Disease ; Adult ; Carcinoma ; radiotherapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Epidermal Growth Factor ; blood ; Female ; Head and Neck Neoplasms ; radiotherapy ; Humans ; Incidence ; Leukocyte Count ; Male ; Middle Aged ; Phytotherapy ; adverse effects ; Platelet Count ; Radiation Injuries ; drug therapy ; prevention & control ; Stomatitis ; drug therapy ; epidemiology ; etiology ; prevention & control ; T-Lymphocyte Subsets ; drug effects ; Treatment Outcome

OBJECTIVETo generalize the clinical use of transsphincteric operation (Mason operation) for rectal lesions.

METHODSClinical data of 120 patients with middle and lower rectal lesions who underwent Mason operation from Aug. 1990 to Aug. 2005 were analyzed retrospectively.

RESULTSThere were 61 cases with villous adenoma including 26 with cancerization, 25 cases with rectal cancer including 16 cases with early rectal cancer, and 17 with submucosal tumor. Of the 103 patients with rectal tumor, 98 underwent partial rectectomy, 5 segmental rectectomy. The postoperative complications included incision infection in two cases (1.6%), fistula in 4 cases (3.3%). Three patients (3.0%) had postoperative local recurrence. 90.2% of the rectal cancer patients (46/51) survived more than five years after Mason operation.

CONCLUSIONMason operation is satisfactory because of good exposure and simple access to the rectum, which is suitable for those lesions that could be locally resected on the mid and low rectum.

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