Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.

To construct knockout vectors containing ampicillin resistant gene and partial sequence of hyaluronidase gene(Hyl)so that Hyl can be knock out by transforming the plasmid into Streptococcus zoopidemics mutans.First,partial sequence of Hyl(Hyl-1)was cloned into the vector of pMD19-T by using DNA of Streptococcus zoopidemics as template,and then a knockout vector pMD19T-SA was constructed,in which Hyl-1 gene was disrupted by inserting ampicillin resistant gene(Amp)from reverse PCR.As expected,the vector was proved to be consisted of Hyl-1-Amp-Hyl-1-pMD19-T.Thereafter,DNA fragment of Hyl-1-Amp-Hyl-1 was subcloned into pBR322 vector,the resulting construct was then checked by PCR and restriction analysis for the proper configuration of the knockout vector pBR322-SA.Both of the knockout vectors were used to transform Streptococcus zoopidemics and one recombinant was obtained in result.From results of PCR and Hyl activity assay,it was indicated that in the recombinant the Hyl gene was disrupted completely.

Amyloidosis is less involved in the respiratory tract. The tracheobronchial amyloidosis is rare, and the amyloid changes involved in the larynx, trachea and bronchus are more rare. The study introduces a case of the primary laryngeal and tracheobronchial amyloidosis. The patient was diagnosed as primary laryngeal amyloidosis, and had tracheobronchial amyloidosis after 15 years. Due to the rare and non-specific clinical manifestations of the disease, the patient was misdiagnosed as upper respiratory tract infection and bronchial asthma.After the treatment of the clamp of the bronchoscope and argon knife,the symptoms were significantly relieved in the patient.At present,the disease is stable and has not relapsed,but it still needs to track the long-term prognosis.

OBJECTIVEIn order to improve the irrigation for Panax notginseng growing seedlings, different mulching ways were carried out to investigate the effects of double mulching.

METHODField experiment was applied to study soil moisture, soil temperature and bulk density of different mulching ways while the germination rate and seedlings growth also were investigated.

RESULTCompared with the traditional single mulching with pine leaves or straw, double mulching using plastic film combined with pine leaves or straw could reduce 2/3 volumes of irrigation at the early seedling time Double mulching treatments didn't need to irrigate for 40 days from seeding to germination, and kept soil moisture and temperature steady at whole seedling time about 30% and 9.0-16.6 degrees C, respectively. The steady soil moisture and temperature benefited to resist late spring cold and germinate earlier while kept germination regularly, higher rate and seedlings quality. In contrast, single mulching using pine leaves or straw had poor soil moisture and temperature preserving, needed to irrigate every 12-day, meanwhile dropped the germination and booming time 14 days and 24-26 days, respectively, reduced germination rate about 11.3%-8.7%. However, single pine leaves mulching was better than straw mulching. In addition, though better effects of soil moisture and temperature preserving as well as earlier and higher rate of germination with single plastic films mulching had, some disadvantages had also been observed, such as daily soil temperature changed greatly, seedling bed soil hardened easily, more moss and weeds resulted difficulty in later management.

CONCLUSIONTo the purpose of saving water and labor as well as getting higher germination rate and seedlings quality, double mulching using plastic films combined pine leaves at the early time and single mulching removing plastic films at the later time is suggested to apply in the growing seedlings of P. notoginseng.

Agriculture ; methods ; China ; Drugs, Chinese Herbal ; analysis ; Fertilizers ; analysis ; Panax notoginseng ; chemistry ; growth & development ; Quality Control ; Seedlings ; chemistry ; growth & development ; Soil ; chemistry

BACKGROUNDTotal hip arthroplasty (THA) in developmental dysplasia of the hip (DDH) is more complex than the normal hip, with large replacement risks and many complications. Although nonosteotomy THA is convenient to perform, femoral osteotomy shortening can avoid blood vessel and nerve traction injuries. This study aimed to compare osteotomy THA with nonosteotomy to determine reasonable options for operative management of DDH.

METHODSData on 48 DDH patients who underwent THA were analyzed retrospectively. The patients were divided into two groups: Group A 29 cases (nonosteotomy), and group B 19 cases (osteotomy). Harris and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, limb length discrepancy (LLD), radiological data on the hip, and claudication were evaluated. Data were analyzed by using paired-sample Student's t-test, independent-sample Student's t-test, and Pearson's Chi-square test; the test level was α =0.05.

RESULTSPostoperative Harris (90.7 ± 5.1) and WOMAC scores (88.0 ± 10.6) were significantly improved compared with preoperative Harris (44.8 ± 5.7) and WOMAC scores (42.0 ± 5.3) in group A (P < 0.05). Postoperative Harris (90.4 ± 2.8) and WOMAC scores (88.2 ± 5.9) were significantly improved compared with preoperative Harris (44.4 ± 4.2) and WOMAC scores (43.2 ± 4.3) in group B (P < 0.05). One case of dislocation occurred in group A; after closed reduction, dislocation did not recur. In group A, 2 patients developed cutaneous branch injury of the femoral nerve, which spontaneously recovered without treatment. Postoperative LLD >2 cm was seen in one case in group A and five cases in group B. Postoperative claudication showed no significant difference between the two groups (P > 0.05). No patients developed infection; postoperative X-rays showed that the location of the prosthesis was satisfactory, and the surrounding bone was not dissolved.

CONCLUSIONSTHA is effective and safe for DDH. For unilateral high dislocation DDH patients with limb lengthening ≤4 cm and good tissue conditions, THA without femoral osteotomy may be considered.

Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Hip Dislocation, Congenital ; surgery ; Humans ; Male ; Middle Aged ; Osteotomy ; methods ; Postoperative Period ; Range of Motion, Articular ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the pulmonary function after treatment in neonates with respiratory distress syndrome (RDS) at varying disease severity levels and different gestational ages.

METHODSA total of 107 neonates with RDS were divided into <34 weeks group (65 neonates), late preterm group (21 neonates), full-term group (21 neonates). Another 121 non-RDS children were enrolled as the control group. According to the severity of RDS, the RDS neonates were divided into mild RDS group (grades 1 and 2; 76 neonates), and severe RDS (grades 3 and 4; 21 neonates). The tidal breathing pulmonary function was measured at a corrected gestational age of 44weeks.

RESULTSThe pulmonary function parameters showed no significant differences across the groups of RDS neonates of different gestational ages; the tidal volume per kilogram of body weight (VT/kg) showed no significant difference between the RDS and non-RDS groups, while the RDS group had significantly higher ratio of time to peak tidal expiratory flow to total expiratory time (tPTEF/tE) and ratio of volume to peak tidal expiratory flow to total expiratory volume (vPTEF/vE) than the non-RDS group of the same gestational age (P<0.05). At a corrected gestational age of 44 weeks, the two groups of neonates with varying severity levels of RDS had significantly lower tPTEF/tE and vPTEF/vE than the control group (P<0.05), and tPTEF/tE and vPTEF/vE tended to decrease with the increasing severity level of RDS.

CONCLUSIONSNeonates with RDS have significantly decreased pulmonary function than those without RDS. At a corrected gestational age of 44 weeks, the tidal breathing pulmonary function in neonates with RDS is not associated with gestational age, but is associated with the severity of RDS.

Female ; Gestational Age ; Humans ; Infant, Newborn ; Lung ; physiopathology ; Male ; Respiratory Distress Syndrome, Newborn ; physiopathology

The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism

Adolescent ; Brain Injuries ; blood ; complications ; Cerebral Infarction ; blood ; etiology ; Child ; Child, Preschool ; Female ; Fibrin Fibrinogen Degradation Products ; analysis ; Humans ; Male

Objective To evaluate the clinical efficacy and safety of anti-platelet-derived growth factor combined with ranibizumab in the treatment of patients with age-rclated macular degeneration.Methods A total of 58 cases of age-related macular degeneration were randomlv divided into control group (29 cases) and treatment group (29 cases).The two groups were treated with intravitreal injection with 10 mg · mL-1 ranibizumab,each time 0.05 mL,1 time a month for 3 months.The treatment group was also orally given levamlodipine besylate 5 mg,1 times a day for 3 months.The patients were followed up for 6 months after treatment.The best corrected visual acuity,central macular thickness,fundus hemorrhage area,fluorescein leakage area,and complication rate were measured and analysed.Results After treatment,the total effective rates in control group and treatment group were 72.41％ (21 cases/29 cases),92.59％ (25 cases/27 cases),with significant difference (P ＜0.05).The total improvement rates in control group and treatment group were 75.86％ (22 cases/29 cases),96.30％ (26 cases/27 cases),with significant difference(P ＜ 0.05).After treatment,the corrected visual acuity of control group and treatment group were 0.27 ± 0.08,0.34 ± 0.06,the thickness of the macular fovea were (419.90 ± 30.91),(395.67 ± 6.49) μm,the area of fundus hemorrhage were (0.31 ± 0.13),(0.22 ± 0.06) mm2,and fluorescein leakage area were (0.95 ± 0.17),(0.86 ± 0.13)mm2,all with significant difference (all P ＜ 0.05).The adverse drug reactions in treatment group and control group were high intraocular pressure,conjunctival bleeding and vitreous floating.The incidence of adverse drug reactions in control group and treatment group were 27.59％ (8 cases/29 cases),22.22％ (6 cases/27 cases),with no significant difference (P ＞ 0.05).Conclusion Anti-platelet-derived growth factor can improve the clinical efficacy of ranibizumab in the treatment of age-related macular degeneration.

To confirm the mechanism of exosomes as tumor vaccines inducing immunity response, dendritic cells (DCs) were induced from human peripheral blood mononuclear cells, while exosomes were isolated from DC loaded tumor antigen. The effect of exosomes on priming T cell proliferation was analysed under conditions with or without DCs, or DCs at different mature stages. The function of exosomes in immunity was detected through block test after blocking some molecules (CD11a, CD11b, CD11c, CD54, MFG-E8 and CD83). The effect of DCs on embedded exosomes was observed by confocal microscopy, the effect of blocking surface molecules on exosomes on DC-embedding exosomes was assayed by flow cytometry. The results indicated that both exosomes derived from imDC (imDex) and exosomes derived from mDC (mDex) could not prime T cells without DC or with imDC. The exosomes derived from mDC induced with different cytokines (LPS, TNF-alpha, CpG, CD40L) were no significant difference in concentrations but were different in effect. The immunity function of exosomes depended on CD11a, CD11b, CD11c, CD54, MFG-E8 and CD83 molecules, the effect of priming T cells is reduced when these molecules were blocked. Confocal microscopy and FACS assay showed that blocking CD11a and CD54 could inhibit exosome-targeted DC and DC-embedded exosomes. It is concluded that the exosomes target DCs through their surface molecules, therefore results in immune response of T cells.
Antigens, Neoplasm ; immunology ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; secretion ; Exosomes ; immunology ; Humans ; K562 Cells ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; cytology ; immunology