ObjectiveTo investigate the relationship between the prognosis and both of treatment way and curative age of congenital muscular torticollis.Methods138 children accepted conservative and/or operative treatment according to different ages.A prospective study had been made. ResultsConservative treatment was given to 82 children,71 of them were <1-year-old. 49 of them were follwed up and had satisfactory outcome.7 patients who was >1-year-old had operations later because of asymmetry of head.63 children received operations.56 of them were followed up over one year,47 cases had good-looking,the outcome of other 9 was not satisfied.Conclusions Operative intervention was necessary for >1-year-old patients. Manipulation was suitable way to cure <1-year-old patients.It was the best operating time at the age of 1-5 years old.

After a literature review of the HPMC capsules, the research status of the HPMC capsules in vivo and in vitro are summarized, including the application status, the superiority over hard gelatin capsules and in particularly the disintegration release in vitro and bioavailability in vivo, as well as pharmacokinetics difference compared with conventional gelatine capsules, are explored in depth. Finally, the future applications of HPMC capsules are prospected.

OBJECTIVE:To prepare Carbinoxamine maleate sustained-release suspension,and evaluate its quality. METH-ODS:Using carbinoxamine maleate as raw material,drug-loaded resin was prepared by cation exchange resin;surface coating method was used to finally prepare sustained-release suspension,using Eudragit RS100 as sustained-release coating material to pre-pare sustained-release microparticles. HPLC was conducted to determine the content of carbinoxamine maleate,release degree of original preparations and self-made suspensions was compared,drug-loading capacity was calculated. RESULTS:The drug amount in preparing drug-loaded resin was 2%,reaction temperature was 25 ℃,and reaction time was 4 h;the drug-loading capacity in surface coating was 35%,amount of coating material was 10%,and reaction temperature was 40 ℃. The drug-loading capacities of sustained particles before and after coating were 35.23%,32.72%,respectively;the yield was 96.82%. The carbinoxamine ma-leate in prepared sustained-release suspension accounted for 98.76% of the labeled amount;release degree in 10 h reached about 80%,f2 was 65.73. CONCLUSIONS:Carbinoxamine maleate sustained-release suspension is prepared successfully,and its release is similar to the original preparation.

Based on the structure of 5-fluoroindol-2-one and fragments from thirteen multi-target tyrosine kinase inhibitors which have been marketed or in the phase of clinical research, eleven 3-aromatic Shiff base-5-fluoroindol-2-one derivatives were designed and synthesized. Their structures were identified by 1H NMR, MS and elemental analysis. In vitro antitumor bioactivities evaluation was done by MTT method. It was shown that most of synthesized compounds had antitumor activities and compounds 1b, 1g, 1i and 1h were better than or equal to the antitumor activity of positive control.

Objective In this article Response Surface Analysis(RSA)was applied to optimize the formulation of doxycycline hydrochloride sustained release tablet.Methods Single factor exploration was used to determine the three factors which have the greatest impact on the release rate.The three factors were the dosage of the HPMC in the total weight of the tablet,the concentration of PVP-K30,and the ratio of lactose to microcrystalline cellulose,respectively.The composite score of the release behaviour was taken as the response value.The dosage of the ingredients were determined by Box-Benhnke design principles and 3 factors and 3 levels.Results The optimized formulation and process are as follows:the dosage of the HPMC in the total weight of the tablet was 30%;the concentration of PVP-K30 was 10%,and the ratio of lactose to microcrystalline cellulose was 13.The release behavior in vitro is ideal.Conclusion The optimized preparation process of doxycycline hydrochloride sustained release tablet is stable,highly efficient and suitable for industrial production.

To prepare the polyclonal antibody against the 3D polymerase of foot and mouth disease virus (FMDV), the 3D polymerase gene of this virus was amplified by PCR and doubly digested with BamH I and Nde I. Then, it were cloned into expression vector pET-30a(+) to obtain the recombinant plasmid pET-3D and this plasmid was transformed to E.coli BL21(DE3) with induction by IPTG.The target protein was identified and purified with SDS+PAGE, and the inclusion bodies were extracted. The purified target protein was used as antigen to immunize New Zealand rabbits to prepare the polyclonal antibody against 3D polymerase of FMDV, which was then characterized by indirect ELISA and Western blotting. As demonstrated by SDS-PAGE, the target protein with a molecular weight at 46 ku was expressed. The polyclonal antibody showed high affinity and obvious specificity and its titer was above 1:8 000. This polyclonal antibody may lay a foundation for the further studies on the biological functions and epitopes of the 3D polymerase of FMDV.

Objective To identity the distribution of enterotoxin and hemolysin,as well as the clonal complexes and drug resistance of the strains of methicillin-resistant Staphylococcus aureus (MRSA) in Maanshan region.Methods Automatic enzyme-linked fluorescent assay system and PCR technology were used to identify the distribution of enterotoxin and hemolysin genes.Seven Staphylococcus aureus hourskeeping genes were choosed as the target genes for multilocus sequence typing (MLST) on 34 strains of MRSA and 3 strains of methicillin-sensitive Staphylococcus aureus (MSSA),comparing the data with the online database and obtaining the sequence typing (ST),conducting affinity analysis on its ST based on eBURST,testing in agar dilution method the drug resistance of MRSA against 12 antibiotics.Results 50.9％ of the 210 Staphylococcus aureus strains were enterotoxin positive,and 97.1％ of them carried hemolysin genes as all 51 strains of MRSA carried hemolsin genes.The 34 MRSA strains were divided into 10 STs,ranging in sequence ST239 (47.1％,16/34),ST5 (17.6％,6/34).Three MSSA strains belonged to ST188,ST1281 and ST7,respectively.Seventeen strains from the patients were divided into 6 STs,ranging in sequence ST239 (35.3％,6/17) and ST5 (29.4％,5/17).Twenty strains from food sources were divided into 9 STs,ranging in sequence ST239 (45.0％,9/20) and ST7 (15.0％,3/20).STs of ST585,ST630 and ST239 were close in affinity,while the rest were distant in affinity.Except for vancomycin,all the strains were found with drug resistance to varying extent to the 10 antibiotics tested.Conclusion Existence of Staphylococcus aureus hemotoxin was universal; ST239 was the main predominant MRSA in Maanshan region,with distant affinity among the STs.

Cysticercosis,caused by the larval stage of the tapeworm Taenia pisiformis,known as Cysticercus pisiformis,is a parasitic ailment affecting lagomorphs,particularly domestic rabbits,posing a threat to the rabbit industry and the safety of rabbit meat products.This study aims to i-dentify the distribution of the TPO18 antigen in Cysticercus pisiformis and Taenia pisiformis and establish an indirect enzyme-linked immunosorbent assay(ELISA)for detecting antibodies against rabbit cysticercosis.The research involved the prokaryotic expression of the 18 kDa antigen of rabbit Cysticercus pisiformis and the isolation of soluble TPO18 protein post-purification.Immuni-zing rabbits with the TPO18 protein resulted in the production of polyclonal antibodies with a titer of up to 1∶51 200.Western blot analysis validated the antigenicity of the polyclonal antibodies a-gainst total proteins from rabbit Cysticercus pisiformis,Taenia pisiformis and the recombinant TPO18 protein.Immunohistochemistry revealed the distribution of the TPO18 antigen in rabbit Cysticercus pisiformis and Taenia pisiform is,indicating the effective reactivity of the polyclonal antibodies with total proteins from both parasites and the recombinant TPO18 protein.TPO18 an-tigen in rabbit Cysticercus pisiformis predominantly localized in the germinal layer and the paren-chyma,while in Taenia pisiformis,it was mainly present in the suckers,sucker peripheries,collec-ting duct upper cells,and parenchyma.An indirect ELISA based on the TPO18 antigen was devel-oped using the recombinant antigen,and its technical parameters were optimized.The optimized ELISA conditions included a serum dilution of 1∶100,antigen coating concentration of 5 mg/L,coating for 1 h at 37 ℃ followed by overnight incubation at 4 ℃,blocking with 1％BSA for 60 min at 37 ℃,serum reaction for 60 min,secondary antibody dilution at 1∶1 000,secondary antibody incubation for 60 min,substrate reacting for 15 min,with a cutoff value of 0.295.Sensitivity,speci-ficity,and repeatability tests of the ELISA demonstrated high sensitivity and specificity without cross-reactivity with positive sera of rabbit hemorrhagic disease virus,Hepatic coccidiosis,Eimer-ia stiedae,or Toxoplasma gondii.The intra-and inter-assay coefficients of variation were both less than 7％,indicating excellent repeatability.Application of this ELISA,compared to postmortem ex-amination,on 86 clinical serum samples showed a concordance rate of 97.7％.In conclusion,this study successfully established an indirect ELISA for detecting antibodies against rabbit Cysticercus pisiformis,presenting a novel monitoring approach for assessing rabbit infections with Cysticer-cus pisiformis.

OBJECTIVEAttempting to isolate and cultivate the microcytin-RR-producing cyanobacteria from natural blooms as well as to further investigate some characteristics of their growth and metabolite toxicity.

METHODSCapillary-pipette method was used to isolate wild Microcystis strains collected from eutrophicated lakes. The isolated strains were cultured in BG11 media at (25 ± 1) °C, under 2 000 lx illumination of fluorescent light with a light-dark rhythm of 12-12 h. The growth curve was observed by measuring optical density of culture suspension, toxin-related genes and the metabolite toxins were identified separately by PCR and HPLC, and its acute toxicity was carried out by orally administered toxins to Kunming (KM) mice.

RESULTSOne of five toxigenic strains from 198 collected samples was confirmed to be a MC-RR producing blue-green alga by existing two specific toxin-synthesized enzyme genes and showing specific chromatographic peak of the toxin compared with standard MC-RR through both PCR and HPLC methods. The toxic strain was classified as Microcystin aeruginosa by morphologic and phylogenetic tree analysis. The growth length of the strain lasted nearly 81 days with 55-60 days' exponential phase and the maximal concentration of 5.52 × 10⁷ cell/ml. The LD50 of the MC-RR to the KM mice ranged from 10.75 mg/kg to 13.45 mg/kg of body weight. As a result of the acute toxicity, the enzymatic indexes in serum such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) were significantly higher than those in the control group. The levels of ALT, AST, ALP and LDH in the treated group at 45 min were (157.08 ± 20.38), (333.00 ± 68.53), (392.70 ± 89.59) and (1 071.13 ± 160.22) U/L respectively, and at 4 h were (514.68 ± 156.87), (593.15 ± 40.41), (618.55 ± 208.76) and (2 281.72 ± 866.67) U/L respectively, and meanwhile the values of ALT, AST, ALP and LDH in the control group were (40.30 ± 4.89), (142.70 ± 26.59), (56.90 ± 11.89) and (509.50 ± 94.75) U/L separately (t values at 45 min were -11.20, -5.77, -7.38, -6.60 respectively, and at 4 h were -6.04, -20.21, -5.35, -4.07 respectively, P values were all <0.01). The liver coefficient in the treated group at 45 min and 4 h were 6.855 ± 0.225 and 8.409 ± 0.276, significantly higher than that (5.784 ± 0.286) in the control group (t values were -3.96 and -12.22, P values were both <0.01). The histopathological changes of liver were hyperemia obviously.

CONCLUSIONIsolated from the bloom waters, a strain of Microcystis aeruginosa is obtained with characteristics of longer growth duration, positive microcystin synthetase genes, and dominant production of MC-RR. The LD50 of the extracted MC-RR administered by oral route to mice is (12.10 ± 1.35) mg/kg of body weight, and liver is the target organ of MC-RR. The existence and potential risk of MC-RR in China cannot be ignored.

Animals ; China ; Chromatography, High Pressure Liquid ; Cyanobacteria ; Hyperemia ; Lakes ; Liver ; Mice ; Microcystins ; Microcystis ; Phylogeny

Feces and intestinal contents of pigs suspected with porcine epidemic diarrhea virus were collected from a farm in Minqin County,Gansu Province,China.After the suspected positive sam-ples were detected by RT-PCR,Vero cells were used to isolate and culture them in vitro.The suc-cessfully isolated virus was identified in the laboratory,and its whole genome sequence was ana-lyzed for genetic evolution.The pathogenicity was evaluated by animal regression test.The results showed that typical syncytial lesions could be observed when the PEDV-positive treatment solu-tion was inoculated with Vero cells in the 4th generation,and the virus titer in the 6th generation reached 10-4 75TCID50/mL.PEDV-like virions with a diameter of about 100 nm and a round shape with obvious capsular membranes and spikes were observed by electron microscopy.Whole genome sequencing analysis showed that the total length of this strain was 28 085 bp,which was far from the G1 subtype represented by the classical strain CV777(96.6％),and had a high homology with the G2b strains BC-2011-1,IA1,USA/Colorado/2013 and WELL(98.6％).This indicated that the strain belonged to the G2b epidemic strain.The animal regression test showed that the 5-day-old piglets developed vomiting,acute watery diarrhea,emaciation and mental depression within 12 h after the attack,and the symptoms worsened and died within 24 h.After autopsy,the infected piglets could be observed with stomach swelling,high intestinal heave,thin and transparent intesti-nal wall,and undigested milk clots inside.In summary,a PEDV G2b epidemic strain was success-fully isolated and identified in this study,and its whole genome sequence and pathogenicity were analyzed,providing research materials for future studies on PEDV gene function,pathogenic mech-anism and vaccine development.