Objective To compare the osseo-induction and biodegradation performances of three types of injectable and degradable calcium phosphate cement (CPC) so as to find out a better bone substitute. Methods Three types of injectable and degradable CPC were respectively implanted into the bilateral tibias of 24 New Zealand rabbits: pure CPC (Group A), CPC added with Zinc and Strontiumions (Group B), and CPC with composite rhBMP-2 (Group C) . Their systematic and local reactions in implanted region were closely observed. The degra- dation and osseo-induction performances were compared macroscopically, microscopically and by CT scan to find out the one that could best meet clinical needs. Tissue slices were sampled and photographed four, eight and 16 weeks after operation. Five photographs were selected in each group and at each time points for computer software (Image Pro Plus 5.1) processing to calculate the percentages of bone in the images of postoperative slices. Results In Groups A and B, new bone was found to form slowly and little by little, and the ossification was not synchronous with the material degradation. In Group C, however, new bone was observed to form early and massively, and the os- sification was almost synchronous with the material degradation. In Groups A, B and C, the percentage of bone in the images of postoperative slices was (41.7?16.6)%, (31. 2?12.2)% and (71.7?21.0)% respectively. The bone percentage in CPC with composite rhBMP-2 was significantly higher than that in the other two types of CPC (P＜0.01 ). Conclusion The injectable and degradable CPC with composite rhBMP-2 is more suitable for clinical use, because it can induce early new bone formation and synchronous biodegradation.

Objective:To provide anatomic image data for ossointegrated implant for patients with nasal defect. Methods: 100 cases of (50 males, 50 females) 3D spiral CT images of normal bone of skull were measured with mimics 10.01 software. Results: The minimum distance between the root tips of central incisor and the anterior nasal floor was 13.03 in males and 11.06 in females. The minimum distance between the root tips of lateral incisor and the anterior nasal floor was 15.95 mm in males and 13.04 mm in females, the minimum thickness of palatine process 6mm inferior to piriform aperture was 9.96 mm, the minimum thickness of alveolus 6mm inferior to piriform aperture was 11.57 mm in males and 10.61 mm in females, the thickness of glabella adjoin to frontal sinus was 3.57mm. Conclusion: The lateral incisor region is an optimal one for nasal implants 4-8 mm in length. The inclined direction can either be forward or backward. When the vertical bone depth is not enough, horizontal placement of the implants in the inferior region of piriform should be considered. 4-8 mm implants is suitable in this region. The glabella region is not optimal but an alternative option for nasal implants.

Objective To investigate the amino acid patterns in penicillin-binding protein 2(PBP2)in Neisseria gonorrhoeae isolates with reduced susceptibility to ceftriaxonc.and the relationship between the amino acid patterns and reduced ceftriaxone susceptibility.Methods DNA was extracted from 13 clinical isolates of N.gonorrhoeae.including 11 strains with decreased susceptibility to ceftriaxone and 2 sensitive isolates.The full-length penA gene encoding the penicillin-binding protein 2 was amplified and sequenced.BLASTn and BLASTx programs were used to assess the insertion and substitution patterns of nucleotides in penA gene and of amino acids in PBP2,respectively.Results BLASTn analysis revealed insertion or substitution of 18-38 nucleotides in the penA gene of gonococcal isolates with reduced ceftriaxone susceptibility.As shown by BLASTX analysis.there were five patterns of amino acid substitution or insertion in PBP2 of the 11 isolates with reduced ceftriaxone susceptibility.However.mosaic structure of PBP2 was not found in any of these isolates.Conclusion Mosaic PBP2 seems not to be the major factor contributing to the decrease in susceptibility of N.gonorrhoeae to ceftriaxone.

Conjugal plasmid pGH112 has been developed based on the replicons of Streptomyces coelicolor plasmid SCP2 and E. coli ColE. The plasmid contains ampicilin resistance gene(amp) for selection in E. coli and thiostrepton resistance gene (tsr) for selection in Streptomycetes, and a 0.76 kb oriT fragment of (IncP) RK2. Conjugal transfer of pGH112 was performed from E. coli to S. coelicolor A3(2), S. avermitilis, S. lividans TK54, S. toxytricini NNRL15443, S. venezuelae ISP5230 and Sacc. erythraea by conjugation, results show that the plasmid was able to transfer efficenctly from E. coli to Streptomycetes, was stably inherited in the recipients. pGH113 was constructed from pGH112 by combining the constitutive ermE promoter with green fluorescent protein gene(gfp).
Ampicillin Resistance ; genetics ; Conjugation, Genetic ; Escherichia coli ; genetics ; Green Fluorescent Proteins ; genetics ; Plasmids ; Streptomycetaceae ; genetics ; Thiostrepton ; pharmacology

Abstract: Objective　To master the condition of cockroach population distribution, seasonal dynamics, cockroach density for different habitat, and to provide a basis for developing cockroach control strategies. Methods　Six types of surveillance sites, including residential areas, hotels, restaurants, supermarkets, hospitals and farm product markets, were set up in 14 cities in Liaoning to monitor cockroaches using the sticky-trap method. The cockroach surveillance data from vector surveillance sites in fourteen cities of Liaoning Province in 2021 were collected and statistically analyzed using Excel 2010 and SPSS 23.0 software. The density and species composition of cockroaches were analyzed, and the density difference and seasonal dynamics trend of cockroaches in different habitats were compared. Results　A total of 3 031 cockroaches were captured in 2021, of which Blattella germanica accounted for 94.66% (2 869/3 031) and was the dominant population. The total density of cockroaches was 0.230 0 cockroaches per sheet (3 031/13 234) and the total infestation rate of cockroaches was 5.59% (562/10 052). The density and infestation rate of cockroaches in different habitats were in the order of farm product markets, restaurants, and hotels and the difference in infestation rate between habitats was statistically significant (χ2=168.327, P<0.05). The seasonal dynamics trend of cockroach density and disoperation rate showed a unimodal curve, and the peaks were distributed in July. The seasonal dynamics of cockroach density and disoperation rate in different habitats showed a unimodal curve in the habitats of farm product markets, supermarkets, hotels, hospitals and residential areas all, while the habitats of restaurants were close to a double peak curve. Conclusions　B. germanica is the dominant species of cockroaches in Liaoning Province in 2021. Compared with 2020 the density and disoperation rate of cockroach in 2021 showed a slight downward trend, and the seasonal dynamics trend of cockroach density and disoperation rate showed a unimodal curve. The farm product markets are the key places for cockroach prevention and control. According to the seasonal fluctuation trends in cockroach density and infestation rate in different habitats, comprehensive prevention and control measures should be taken before the peak periods to reduce cockroach density and control diseases.

OBJECTIVETo establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay.

METHODSRat kidney samples were hydrolyzed in 6 mol x L⁻¹ HCl under 110 degree for 24 h, then 9-fluorenylmethyl chloroformate (FMOC) was added for the pre-column reaction. The HPLC analysis was performed on a Phenomenex C₁₈ column using sodium acetate buffer, methanol and acetonitrile as mobile phase with a flow rate of 1.0 ml min⁻¹ and UV detective wavelength 265 nm at 40 degree.

RESULTSLinear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%.

CONCLUSIONThe established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.

Animals ; Chromatography, High Pressure Liquid ; methods ; Hydroxyproline ; analysis ; In Vitro Techniques ; Kidney ; chemistry ; Male ; Rats

To study the influence of hypercholesterolemia with caveolin-1 on the plasmalemma of vascular endothelium, we used the methods of immunohistochemistry to detect the dynamic changes of caveolin-1 in cultured ECV-304 cells which were stimulated high cholesterol serum and the arterial endothelium of hypercholesterolemia rats. It is resulted that high cholesteorol level can upregulate the expression of caveolin-1 both in vitro and in vivo. In the initial stage of hypercholesterolemia model, the expression of caveolin-1 increased as the time of high cholesterol level added, but in the later period it was decreased slightly.
Animals ; Aorta ; pathology ; Caveolin 1 ; Caveolins ; biosynthesis ; genetics ; Cells, Cultured ; Cholesterol ; blood ; Endothelium, Vascular ; cytology ; metabolism ; Female ; Humans ; Hypercholesterolemia ; metabolism ; Male ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Umbilical Veins ; cytology

In order to provide the evidences for CD19 as a better antibody targeting molecule for B lineage acute leukemias than CD20 through the multi-parameter flow-cytometry analysis of leukemia cells, the samples from 321 patients with acute leukemia (AL) were immunophenotyped by multi-color flow cytometry and CD45/SSC gating strategy followed by the analysis of CD19 and CD20 expression. The results showed that the positive rate of CD19 (115/116, 99.1%) in 116 cases with B lineage acute lymphoblastic leukemia (B lineage ALL) was significantly higher than that of CD20 (33/116, 28.4%) (P < 0.01); in 17 patients with B lineage/Myeloid (B/My) acute mixed lineage leukemia (AMLL), the former positive rate (17/17, 100%) was also higher than the latter (5/17, 29.4%) (P < 0.01). Both of the two antigens were negative in 29 patients with acute T lymphoblastic leukemia and 7 patients with T/My AMLL. The positive rates of CD19 and CD20 in 152 patients with acute myeloid leukemia (AML) were 7.2% and 2.0%, respectively. The difference of the fluorescence intensity between the two antigens on the cells from each patient with B lineage ALL or B/My AMLL was statistically significant (t = 20.68, P < 0.001). The specificity of CD19 and CD20 in B lymphocytic lineage was 92.3% (132/143) and 92.7% (38/41), respectively, while the sensitivity was 99.2% (132/133) and 28.6% (38/133), respectively, the former sensitivity was significantly higher than the latter (chi(2) = 144.018, P = 0.001). It is concluded that CD19 continuously and steadily express on almost all subtypes of B lineage leukemic cells with homogeneous pattern while only a small number of leukemias express CD20. Both the specificity and sensitivity of CD19 were very high with a much broader reaction pattern than that of CD20 on this group of diseases. These indicate that CD19 may be a better antibody targeting molecule than CD20 for patients with B-lineage acute leukemia.
Acute Disease ; Adolescent ; Adult ; Antigens, CD19 ; biosynthesis ; Antigens, CD20 ; biosynthesis ; Bone Marrow Cells ; immunology ; metabolism ; pathology ; Cell Lineage ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Infant ; Leukemia ; blood ; immunology ; pathology ; Leukocytes, Mononuclear ; immunology ; metabolism ; pathology ; Male ; Middle Aged ; Tumor Cells, Cultured

OBJECTIVETo construct a prokaryotic vector of ZCH-7-2F9 single chain antibody (ScFv2F9) and to obtain the ScFv2F9 protein with biological activity for further studies.

METHODSPrimers were synthesized according to the gene sequence of ScFv2F9, four tandem glycin and one serine (G4S) 3linker and multiple cloning site(MCS) of pIVEX2.3-MCS vector, which included NdeI and SmaI enzyme cleaving sites. ScFv2F9 gene was amplified through splicing by overlap extension (SOE) using the high fidelity Taq polymerase. Then the gene was cloned to pGEM-T easy and pIVEX2.3-MCS vectors. Positive recombinants (pIVEX2.3-MCS/ScFv2F9) were identified through enzyme cleaving and sequenced before expression. The recombinant plasmids were transfected into E.coli BL21star(DE3)plysS for expression. After purification with Ni+resin and renaturation in vitro, the relative molecular mass (Mr) and the binding activity of the interesting protein were determined by SDS-PAGE and flow cytometry (FCM), respectively.

RESULTThe cloned ScFv2F9 gene was identified to be functional by sequencing and expressing. The interesting protein was detected in inclusion body with a Mr of 31 000. The blocking test showed that the positive cell percentage, the mean fluorescence intensity (MFI) and the peak of channel (peak Ch) were reduced by 11.73%, 11.96% and 31.57%, respectively after once blockage by scFv2F9 protein, and 26.44 %, 21.75 % and 42.11 % after blockage twice.

CONCLUSIONThe ScFv2F9 against human CD14 antigen has been successfully expressed in prokaryotic cells with partial biological activity, which lays the foundation for further studies on its immunotoxin and other kinds of engineered antibodies.

Antibodies, Monoclonal ; Biotechnology ; methods ; Cells, Cultured ; Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Humans ; Lipopolysaccharide Receptors ; immunology ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; genetics

Objective　To investigate the actions of extra cellular medium in growth and differentiation of hair follicle and to look for growth adjusting factors for dermal papilla cells (DPC). Methods　Dermal papilla cells were isolated and cultivated with two steps method and the cells were identified by immunohistochemical staining for actin. Influence was examined on the adhesion and growth of dermal papilla cells by chondroitin sulfate A, chondroitin sulfate C and heparin sulfate. Results　Two steps method of enzyme digestion for isolating and cultivating dermal papilla cells was an efficient method and large amount of dermal papilla of high purity were harvested with this method. The method is very simple and easy to manege with. Increased adhesion and growth of dermal papilla cells were observed in specimen treated with chondroitin A and heparin sulfate. No significant effects was observed in the cells treated with chondroit in sulfate C. Conclusion　Some extra cellular medium can regulate the adhesion and growth of dermal papilla cells and therefore influence the growth and development of hair follicle.