[Comparative study on extraction of impacted mandibular wisdom teeth by Er:YAG laser and turbine handpiece].

PURPOSE: To compare the clinical effect of Er:YAG laser and turbine handpiece in the removal of lower horizontally impacted wisdom teeth, and to evaluate the operation time, postoperative pain, facial swelling, degree of mouth opening limitation and complications. METHODS: From March 2020 to May 2022, forty patients with bilateral lower mandibular horizontally impacted wisdom teeth in the Department of Oral and Maxillofacial Surgery of Linyi People's Hospital were selected，and all bilateral wisdom teeth were partially bone buried. The bilateral wisdom teeth of each patient were removed by Er:YAG laser on one side and turbine handpiece on the other side, respectively. The patients were divided into experimental(laser) group and control(turbine handpiece) group according to the ways of bone removal on each side. The clinical effect of the two groups was compared after a week of follow-up. Statistical analysis was performed with SPSS 19.0 software package. RESULTS: There was no significant difference between the two groups in operation time(P＞0.05). The incidence of postoperative pain, facial swelling, mouth opening limitation and complications in the experimental group were significantly lower than those in the control group(P＜0.05). CONCLUSIONS: The operation time of extraction with Er:YAG laser is similar to that with turbine handpiece, but laser can reduce postoperative reaction and the incidence of complications, which is easy to be accepted by patients and worthy of wide application.

A crayfish ALF inhibits the proliferation of microbiota by binding to RPS4 and MscL of E. coli.

Antimicrobial peptides (AMPs), most of which are small proteins, are necessary for innate immunity against pathogens. Anti-lipopolysaccharide factor (ALF) with a conserved lipopolysaccharide binding domain (LBD) can bind to lipopolysaccharide (LPS) and neutralize LPS activity. The antibacterial mechanism of ALF, especially its role in bacteria, needs to be further investigated. In this study, the antibacterial role of an anti-lipopolysaccharide factor (PcALF5) derived from Procambarus clarkii was analyzed. PcALF5 could inhibit the replication of the microbiota in vitro and enhance the bacterial clearance ability in crayfish in vivo. Far-western blot assay results indicated that PcALF5 bound to two proteins of E. coli (approximately 25 kDa and 15 kDa). Mass spectrometry (MS), far-western blot assay, and pull-down results showed that 30S ribosomal protein S4 (RPS4, 25 kD) interacted with PcALF5. Further studies revealed that another E. coli protein binding to PcALF5 could be the large mechanosensitive channel (MscL), which is reported to participate in the transport of peptides and antibiotics. Additional assays showed that PcALF5 inhibited protein synthesis and promoted the transcription of ribosomal component genes in E. coli. Overall, these results indicate that PcALF5 could transfer into E. coli by binding to MscL and inhibit protein synthesis by interacting with RPS4. This study reveals the mechanism underlying ALF involvement in the antibacterial immune response and provides a new reference for the research on antibacterial drugs.

Effect of marker segregation distortion on high density linkage map construction and QTL mapping in Soybean (Glycine max L.).

Marker segregation distortion is a natural phenomenon. Severely distorted markers are usually excluded in the construction of linkage maps. We investigated the effect of marker segregation distortion on linkage map construction and quantitative trait locus (QTL) mapping. A total of 519 recombinant inbred lines of soybean from orthogonal and reciprocal crosses between LSZZH and NN493-1 were genotyped by specific length amplified fragment markers and seed linoleic acid content was measured in three environments. As a result, twenty linkage groups were constructed with 11,846 markers, including 1513 (12.77%) significantly distorted markers, on 20 chromosomes, and the map length was 2475.86 cM with an average marker-interval of 0.21 cM. The inclusion of distorted markers in the analysis was shown to not only improve the grouping of the markers from the same chromosomes, and the consistency of linkage maps with genome, but also increase genome coverage by markers. Combining genotypic data from both orthogonal and reciprocal crosses decreased the proportion of distorted markers and then improved the quality of linkage maps. Validation of the linkage maps was confirmed by the high collinearity between positions of markers in the soybean reference genome and in linkage maps and by the high consistency of 24 QTL regions in this study compared with the previously reported QTLs and lipid metabolism related genes. Additionally, linkage maps that include distorted markers could add more information to the outputs from QTL mapping. These results provide important information for linkage mapping, gene cloning and marker-assisted selection in soybean.

PcLys-i3, an invertebrate lysozyme, is involved in the antibacterial immunity of the red swamp crayfish, Procambarus clarkii.

Antimicrobial peptides (AMPs) play important roles in innate immunity against pathogens and lysozymes are a particularly type of AMP. Lysozymes are hydrolytic enzymes that are characterized by their ability to cleave the beta-(1,4)-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in peptidoglycan, which is the major bacterial cell wall polymer. In this work, a lysozyme was identified from Procambarus clarkii and designated PcLys-i3. Quantitative RT-PCR was used to analyze the tissue distribution and expression profiles of PcLys-i3. PcLys-i3 was present in all tested tissues and had high expression levels in gills, stomach and intestine. The expression levels of PcLys-i3 were up-regulated in gills and intestine after challenge with Vibrio parahaemolyticus, Staphylococcus aureus and Aeromonas hydrophila. PcLys-i3 and PcFer proteins can enhance the bacterial elimination in crayfish, whereas the bacterial elimination was weakened when the expression level of PcLys-i3 or PcFer RNAs was suppressed by RNAi. Recombinant PcLys-i3 and PcFer significantly reduced the mortality of crayfish with bacterial infections. Further study found that PcLys-i3 could interact with PcFer in vitro. Finally, the PcLys-i3 and PcFer proteins could bind to bacteria and inhibit bacterial replication. These results suggest that both PcLys-i3 and PcFer play important roles in the antibacterial immunity of red swamp crayfish.

Three STATs are involved in the regulation of the expression of antimicrobial peptides in the triangle sail mussel, Hyriopsis cumingii.

Janus kinase (Jak) and signal transducers and activators of transcription (STAT) signaling pathway is associated in antiviral and antibacterial immune response. Previous studies primarily investigated the function of STATs in mammals. For most invertebrates, only one STAT was found in each species, such as STAT92E was found in Drosophila melanogaster. The studies, which focus on the functional difference between various STATs in the same species of invertebrate, are limited. In the present study, three STATs (HcSTAT1, HcSTAT2 and HcSTAT3) were identified in triangle shell pearl mussel, Hyriopsis cumingii. Phylogenetic analysis showed that HcSTAT1 and HcSTAT3 were clustered with Homo sapiens STAT5, and HcSTAT2 was clustered with Pinctada fucata STAT and Crassostea gigas STAT6. All three STATs could be detected in all tested tissues (hemocytes, hepatopancreas, gill, mantle and foot), and were induced expression when challenged with Staphylococcus aureus or Aeromonas hydrophilia in hemocytes and hepatopancreas. HcSTAT1 regulated the expression of HcDef, HcWAP, HcThe and HcTNF. The expression of HcWAP and HcTNF was down-regulated in HcSTAT2-RNAi mussel. And HcSTAT3 affected the expression of HcTNF. The study is the first report of different functions in antibacterial immune responses between STATs in mollusks.

The role of ficolin-like protein (PcFLP1) in the antibacterial immunity of red swamp crayfish (Procambarus clarkii).

In invertebrates, ficolin-like proteins (FLPs) play important roles in innate immunity against pathogens. Previous studies primarily investigated the functions of FLPs in immune recognition, activation, and regulation. However, limited research has examined the functions of FLPs as immune effectors. In this work, a ficolin-like protein was identified in red swam crayfish (Procambarus clarkii) and designated as PcFLP1. Quantitative RT-PCR and western blot were employed to analyze the distribution and expression profiles of PcFLP1 in the tissues of the crayfish. The results indicated that PcFLP1 was present in all tested tissues, including hemocytes, heart, hepatopancreas, gill, stomach, and mid-intestine. The expression level of PcFLP1 was up-regulated in hemocytes, hepatopancreas and mid-intestines of the crayfish challenged with Vibrio parahaemolyticus. Further study demonstrated that PcFLP1 could protect the hepatopancreatic cells of crayfish from V. parahaemolyticus infection. The recombinant PcFLP1 enhanced bacterial elimination in crayfish, whereas the antibacterial action was inhibited after PcFLP1 was knocked down. Furthermore, PcFLP1 could bound to bacteria and inhibited bacterial replication. These results demonstrated that PcFLP1 plays an important role in the anti-Vibrio immunity of red swamp crayfish.

Determination and toxicokinetics comparison of ketamine and S(+)-ketamine in dog plasma by HPLC-MS method.

A simple and reproducible method was developed for the quantification of ketamine and S(+)-ketamine in dog plasma using a high-performance liquid chromatography system coupled to a positive ion electrospray mass spectrometric analysis. Solid-phase extraction was used for extracting analytes from dog plasma samples. The analytes were separated on a Zorbax SB C(18) column (100 x 2.1 mm, 3.5 microm) with acetonitrile-formate buffer (10 mM ammonium formate and 0.3% formic acid) (17 : 83, v/v) as mobile phase at a flow-rate of 0.2 mL/min. Detection was operated under selected ion monitoring mode. [M + H](+) at m/z 238 for ketamine and S(+)-ketamine and [M + H](+) at m/z 180 for phenacetin (internal standard) were selected as detecting ions, respectively. The method was linear in the concentration range 51.6-2580 ng/mL. The intra- and inter-day precisions (RSD %) were within 11.3% and the assay accuracies ranged from 80.0 to 101.4%. Their average recoveries were greater than 91.1% at all test concentrations. The analytes were proved to be stable during all sample storage, preparation and analysis procedures. The method was successfully applied to the toxicokinetics study and comparison of ketamine and S (+)-ketamine following intravenous administration to dogs.

A rapid resolution liquid chromatographic method for fingerprint analysis of raw and processed caowu (Aconitum kusnezoffii).

A sensitive and reliable rapid resolution liquid chromatographic (RRLC) method coupled with diode array detection has been developed for the fingerprint analysis of raw and processed caowu (Aconitum kusnezoffii). The RRLC fingerprints were established with a Zorbax Extend C18 analytical column (4.6 x 50 mm, 1.8 microm) and gradient elution. Analysis run time was <10 min. The method displayed good precision, stability, and repeatability in fingerprint analysis. Characteristic RRLC fingerprints of caowu were generated and used to assess the consistency and differences in the products. Raw and processed caowu from different sources were analyzed under the developed RRLC conditions, yielding contrasting RRLC fingerprints. Comparison of the RRLC fingerprints of processed and raw caowu indicated that the major constituents changed during processing. Meanwhile, a peak area ratio analysis method was applied to assess their chromatographic fingerprints. In characterizing the constituents of caowu, 11 major chromatographic peaks were identified by mass spectrometry and compared with reference standards and reference data. In summary, RRLC fingerprinting is a rapid and useful way to evaluate the quality of raw and processed caowu.

LC-MS determination and pharmacokinetic studies of paeonol in rat plasma after administration of different compatibility of Su-Xiao-Xin-Tong prescriptions.

A rapid and specific liquid chromatographic/electrospray ionization mass spectrometric (LC/ESI-MS) method has been developed and validated for the identification and quantification of paeonol in rat plasma. Paeonol and internal standard were isolated from plasma samples by liquid-liquid extraction with chloroform. The chromatographic separation was accomplished on a Zorbax-SB C18 column (100x2.1 mm, 3.5 microm). The mobile phase consisted of acetonitrile and 0.1% aqueous formic acid (64:36) was delivered at a flow rate of 0.2 mL/min. Detection was performed on a single quadrupole mass spectrometer by selected ion monitoring mode via electrospray ionization source. Linearity was established for the range of concentrations 0.0525-15.8 microg/mL with a coefficient correlation (r) of 0.9995. The intra- and inter-day precision (RSD%) was lower than 9.34% and accuracy ranged from 93.7 to 102.3%. The lower limit of quantification was 0.0525 microg/mL. The proposed method was used to determine the concentration of paeonol for pharmacokinetic studies. The pharmacokinetics of different compatibility prescriptions of Su-Xiao-Xin-Tong were studied and compared.