The low recovery of pertussis toxin (PT) and the low resolving efficiency of chromatography, due to the instability of PT in low salt condition, are the main challenges for its purification. We aplied 2 mol/L urea to prevent the aggregation and disassociation of PT during the purification by ion-exchange chromatography (IEC) and gel filtration chromatography (GFC). The effect of urea on the purification of PT was studied by ELISA assay and non-reduced SDS-PAGE. The activity recoveries of PT and filamentous hemagglutinin (FHA) in IEC and GFC, the resolution efficiency in GFC and the purities of PT and FHA were improved obviously by adding 2 mol/L urea in the mobile phase. The results highlight the potential application of urea in the acellular pertussis vaccine (APV) manufacture procedure.
Adhesins, Bacterial ; isolation & purification ; Chromatography, Ion Exchange ; methods ; Humans ; Pertussis Toxin ; isolation & purification ; Pertussis Vaccine ; chemistry ; isolation & purification ; Solutions ; Urea ; chemistry ; Vaccines, Acellular ; chemistry ; isolation & purification ; Virulence Factors, Bordetella ; isolation & purification

Objective To discuss the effect of rehabilitation exercise program on surgical patients with prolapse of lumbar intervertebral disc.Methods Rehabilitation exercise program was formulated.70 patients with prolapse of lumbar intervertebral disc who had enestration operation or hemilaminectomy discectomy were randomly divided into the observation group and the control group,each with 35 cases.The observation group had rehabilitation exercise program for intervention,while the control group only received materials including written rehabilitation exercise methods and postoperative announcements to guide self-exercise.Chinese Medical Association of Orthopaedics was used to evaluate the excellent rate of clinical effect,and SF-36 was used to compare patients' health related quality of life before and one year after the surgery.Results 61 out of 70 patients finished the follow-up for a year,in which 31 were the observation group and 30 were the control group.9 cases lost contact,with the rate of 12.8％.The excellent rate of curative effect was 93.5％ in the observation group and 73.3％ in the control group,and the difference was statistically significant (x2 =4.063,P ＜0.05).There was no statistically significant difference in SF-36 scores before surgery (P ＞ 0.05).But one year after the surgery,the score of physiological function,psychological function,body pain,overall health,vitality,social function,emotional function and mental health in SF-36 was respectively (55.4 ± 11.2),(49.6 ± 7.5),(78.1±10.5),(67.5±14.5),(72.3±12.2),(77.0±18.1),(68.6±23.2) and (78.9±10.3) in the observation group,all of which were higher than those in the control group,and the differences were statistically significant (t =2.643,4.321,3.012,2.102,2.412,2.210,2.120,2.321,respectively; P ＜ 0.05).Conclusions Rehabilitation exercise program for surgical patients with prolapse of lumbar intervertebral disc can help improve their clinical effect and postoperative quality of life obviously,which is worthy of clinic application.

This paper summarized nursing measures of a patient with secondary hemochromatosis complicated with multiple organic lesions. As the result of defetoxamine treatment and nursing measures included postural care, movement guidance, observation of vital signs, low iron diet, skin care, liver treatment care,psychological care and discharged guidance, patient's blood serum iron returned to normal level and discharged from hospital.

Objective To explore the adverse reactions of Infliximab and its relevant nursing countermeasures. Methods The clinical data from 13 patients were collected. Condition of adverse reaction was observed and nursing countermeasures were conducted. Results A total of 65 times of Infliximab-infusion were conducted in the 13 patients. Serum sickness reaction appeared 6 times, infection occurred 7 times and liver dysfunction occurred 2 times. According with those adverse reactions, nursing interventions including standard medication,infection prevention and observation were conducted. Conclusions The highest incidence of all adverse reactions is infection followed by serum sickness reaction. The key of nursing care is prevention and care of infection, observation of serum sickness reaction and liver function in order to insure effectiveness and safetyduring treatment.

Objective To evaluate the effects of the anticipated health education plan in patients with prolapse of lumbar intervertebral disc.Methods The anticipated health education plan was established.The patients with window treatment of lumbar disc herniation or lumbar disc herniation and a half laminectomy discectomy were randomly divided into two groups, patients in treatment group were treated with the anticipated health education plan and patients in control group were treated with the conventional health education.The standard-reaching rate of health education and the satisfaction of the patients were investigated at the day before their discharge.And the results were proposed as the evaluation index for estimate of education effect.To compare the health-related quality of life, the SF-36 was completed by the patients before and after surgery at least 6 months.Results In the observed group,the standard-reaching rate of health education was 100.00%,the control group was 74.19%, there were significant differences between the two groups (x2 = 7.032, P ＜0.01); the patient' s satisfaction was 100.00% in the observed group,80.65% in the control group,there were significant differences between the two groups (x2 = 4.613, P ＜ 0.05).The scores of SF-36 were no significant difference between the two groups before surgery (P ＞ 0.05); The total effective rate of observed group was significantly higher than that on control groups with statistical significance (P ＜ 0.05).Conclusions The anticipated health education plan can satisfy the patients, continually improve nursing quality and the life quality of the patients obviously, which is worthy of clinic application.

Objective To indentify differentially expressed genes in recurrent nasopharyngeal carcinoma(rNPC) by DNA microarrays,and analyze chromosomal lecalizations and molecular function by bioinformatics. Methods The primary nasopharyngeal carcinoma (pNPC) tissue samples and rNPC tissue samples were selected, and Affymetrix Gene1.0 ST gene chips were used to indentify differential expressed genes in rNPC, and the bioinformatics was used to analyze their chromosomal localizations as well as molecular functions. Results A total of 44 genes were identified to be differential expressed in rNPC.Thirty-six genes were down regulated, 8 genes were up regulated. Functional classification of down-regulation genes showed that most genes (10 genes,27.8%) belonged to the enzyme activity genes, followed by calcium ion binding genes (7 genes, 19.4% ), protein binding genes (5 genes, 13.9% ), receptor activity genes (4 genes, 11.1% ), ATP binding genes (2 genes, 5.6%), transcription factor genes(2 genes,5.6%), extracellular matrix binding and growth factor binding have 1 gene respectively (each accounted for 2.8%). In addition, the functions of 4 genes (11.1%) were unknown.Functional classification of up-regulation genes showed most genes (3 genes, 37.5%) were unknown, followed enzyme activity genes (2 genes, 25.0%) ,receptor activity, calcium ion binding and voltage-gated ion channel activity genes have 1 genes respectively (each accounted for 12.5% ). These genes were localized randomly on the most the chromosomes, with a majority of them localized on chromosomes 1,17. Chromosome 1 contained the most differentially expressed genes (10, 22.7% ), followed by chromosomes 17(5,11.3%). Conclusions The differential expressed genes in rNPC were supposed to be randomly distributed on most chromosomes, but the majorities were found on chromosomes 1, 17. Abnormality in three groups of genes, including in enzyme activity, calcium ion binding and protein binding associate genes, might play important roles in rNPC. Those genes need to be further studied.

OBJECTIVES: Chronic rhinosinusitis (CRS) is common disease in otorhinolaryngology and will lead to lower airway abnormality. However, the only lung function in CRS patients and associated factors have not been much studied. METHODS: One hundred patients with CRS with nasal polyps (CRSwNP group), 40 patients with CRS without nasal polyps (CRSsNP group), and 100 patients without CRS were enrolled. The difference in lung function was compared. Meanwhile, CRSwNP and CRSsNP group were required to undergo a bronchial provocation or dilation test. Additionally, subjective and objective outcomes were measured by the visual analogue scale (VAS), 20-item Sino-Nasal Outcome Test (SNOT-20), Lund-Mackay score, Lund-Kennedy endoscopic score. The correlation and regression methods were used to analyze the relationship between their lung function and the above parameters. RESULTS: The forced expiratory volume in 1 second (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75) of CRSwNP group were significantly lower than other groups (P<0.05). On peak expiratory flow, there was no difference between three groups. In CRSwNP group, FEV1 was negatively correlated with peripheral blood eosinophil count (PBEC) and duration of disease (r=–0.348, P=0.013 and r=–0.344, P=0.014, respectively), FEF25-75 negatively with VAS, SNOT-20 (r=–0.490, P=0.028 and r=–0.478, P=0.033, respectively) in CRSsNP group. The incidence of positive bronchial provocation and dilation test was lower in CRSwNP group (10% and 0%, respectively), with both 0% in CRSsNP group. The multiple linear regression analysis indicated that change ratio of FEV1 before and after bronchial provocation or dilation test were correlated with PBEC in CRSwNP group (β=0.403, P=0.006). CONCLUSION: CRS leading to impaired maximum ventilation and small airway is associated with the existence of nasal polyp. Lung function impairments can be reflected by PBEC, duration, VAS, and SNOT-20. In CRSwNP patients, PBEC is independent predictor of FEV₁ change ratio.
Bronchial Hyperreactivity ; Bronchial Provocation Tests ; Eosinophils ; Forced Expiratory Volume ; Humans ; Incidence ; Linear Models ; Lung* ; Nasal Polyps ; Otolaryngology ; Ventilation ; Vital Capacity

OBJECTIVETo observe the clinical efficacy of Penyanqing Capsule (PYQC) in treating pelvic inflammation of Qi-stagnation with blood stasis syndrome.

METHODSThe randomized, single blinded, parallel positive drug controlled method was adopted, with 82 patients assigned into two groups by envelop method. The 42 patients in the treated group received PYQC 3 times a day, 4 capsules each time taken orally; the 40 patients in the control group were given orally Fuyankang tablets (FYKT) 3 times a day, 6 tablets each time. The therapeutic course for both groups was 2 months, and 2 courses of treatment were given successively to observe the comprehensive effect, changes of symptoms and signs before and after treatment. The effects of PYQC on hemorrheological character in part of the patients and on the pathogenetic chlamydia and mycoplasma were also observed.

RESULTSThe total effective rate in the treated group was 83.3%, which was insignificantly different from that in the control group (77.5%, P > 0.05). However, PYQC could significantly lower the hemorrheologic indexes in patients and showed definite influence on the pathogenetic chlamydia and mycoplasma.

CONCLUSIONPYQC has good therapeutic effect in treating chronic pelvic inflammation of Qi-stagnation with blood stasis syndrome, and showed definite effect on chlamydia and mycoplasma.

Adolescent ; Adult ; Blood Circulation ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Pelvic Inflammatory Disease ; drug therapy ; Qi ; Single-Blind Method

A new hydrophobic absorbent based on homemade highly cross-linked agarose beads was synthesized by immobilizing butyl derivative onto the matrix linkage. The density of ligand was controlled by adjusting the concentration of butanethiol and the synthesis route was optimized by evaluating the purification efficiency of recombinant Hepatitis B surface antigen (HBsAg) expressed by Chinese hamster ovary (CHO) cell line. A high performance absorbent was finally screened out with up to 80% of HBsAg recovery and purification-fold (PF) about 20. Furthermore, the column pressure was about 0.06 MPa under the flow rate of 500cm/h, and no leaked butyl were detected after exposing the gel in common buffers, chaotropic agents, high concentrations of denaturing agents such as guanidine hydrochloride, urea and polar organic solvents. These results demonstrated that the absorbent have high physico-chemical stability, so it was available for the downstream process. Finally, after scaled up to 2L wet gel/batch, the absorbent was applied to the integration of three-step chromatography and obtained the purified CHO-HBsAg with 95% purity by SDS-PAGE and HPLC, which meet the requirements of SFDA. The purification efficiency and the reproducible ability of the absorbents were also evaluated from batch-to-batch. The results demonstrated that the absorbent met the requirement of scalable, reproducible, economic effect as well. This absorbent is a promising alternative exported HIC gel for wildly being used in Chinese pharmaceutical industries.
Animals ; CHO Cells ; Chromatography, Agarose ; methods ; Cricetinae ; Cricetulus ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; isolation & purification ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

The dissociation of virus-like particles of Hepatitis B surface antigen (HBsAg) during the adsorption-desorption on the solid-phase of chromatography is a main challenge for its purification. Herein, poly (ethylene glycol) (PEG) was applied as an additive during the purification of HBsAg from recombinant Chinese hamster ovary (CHO) cell culture to improve the HBsAg recovery and protect its structural assembly. The presence of 1% of PEG10000 in the mobile phase of ion-exchange chromatography (IEC) of DEAE-Sepharose FF column could increase the recovery of HBsAg from about 55% to 80%, with a similar purification (-fold) (about 12) compared with the absence of PEG. Importantly, glycosylated protein forms of HBsAg were reserved well by PEG-accompanied chromatography. Furthermore, size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) analysis was performed on line to monitor the aggregates, particle size and molecular weight distribution of HBsAg. The results demonstrated that the HBsAg particle size and assembly are more homogenous after adding PEG in the mobile phase of IEC than no PEG added in the mobile phase.
Animals ; CHO Cells ; Chromatography, Ion Exchange ; methods ; Cricetinae ; Cricetulus ; Hepatitis B Surface Antigens ; genetics ; isolation & purification ; Humans ; Polyethylene Glycols ; chemistry ; Recombinant Proteins ; genetics ; isolation & purification