Objective To establish HPLC specific chromatogram of Sendeng-4,a Mongolian medicine compound,and to simultaneously determine six index components—gallic acid,genipin-1-β-D-gentiobio-side,geniposide,(2R,3R)-dihydromyricetin,rutin,andmyricetin.Methods HPLCspecificchro-matogram was obtained under the following system:Extend-C18 column (250 mm ×4.6 mm,5 μm), mobile phase of acetonitrile (A)and 0.6 % phosphoric acid aqueous solution (B),gradient elution, flow rate at 1 mL/min,the column temperature at 25 ℃,detection wavelength at 254 nm,and the injec-tion volume 10 μL.HPLC system for determination was listed as the following:Extend-C18 column (250 mm ×4.6 mm,5 μm),mobile phase of acetonitrile (A)and 0.6%phosphoric acid aqueous solution (B),gradient elution,flow rate at 0.5 mL/min,the column temperature at 27 ℃,the detection wave-lengthsat254nmand292nm,andtheinjectionvolume10μL.Results Morethan90peakswerede-tected on HPLC specific chromatogram,from which ten were identified by comparing with standard HPLC chromatogram and six were measured.After determination,the content and the peak area of each compo-nent had a good linear relationship,with correlation coefficients 0.999 9,average recovery of 98.29%~103.75%andRSD1.44%~2.97%.Conclusion The method established is accurate,simpleandre-peatable,and can be used to control the quality of Mongolian medicine compound Sendeng-4.

Objective To establish the characteristic chromatograms of Mongolian compound Sengdeng-4 in vivo and in vitro, analyze and identify its chemical components in vitro and serum. Methods After the model of rheumatoid arthritis was established, SD rats were orally administered Mongolian compound Sengdeng-4 and each single ingredient respectively. The medicinal serum were separated and measured with HPLC. The HPLC characteristic chromatograms of Mongolian compound Sengdeng-4 in vitro and medicinal serum were identified. The characteristic chromatograms of Mongolian compound Sengdeng-4, single ingredient in vitro and medicinal serum were analyzed, and the chemical composition of Mongolian compound Sengdeng-4 in vitro, original crude medicinal in blood were identified and compared. Results The results of HPLC characteristic chromatograms of Mongolian compound Sengdeng-4 showed that 47 peaks were identified. There were 14 chemical compositions absorbed into blood, among them 9 were original ingredients and 5 were metabolites. The second peak was from Wenguanmu; the fifth, sixth, and seventh peaks were from Zhizi; the first, eighth, and tenth peaks were from Hezi. The third peak was from Wenguanmu, Chuanlianzi and Hezi. The fourth and thirteenth peaks were from Mongolian compound Sengdeng-4. The ninth peak was from Wenguanmu and Chuanlianzi. The eleventh, twelfth
and fourteenth peaks were from Wenguanmu and Hezi. 7 components of Mongolian compound Sengdeng-4 were identified. Conclusion The chemical compositions and metabolites in blood may be the active ingredients with direct actions in vivo, which come from the ingredients of Mongolian compound Sengdeng-4 .

Objective:To observe the effects of modified Taohong Siwu Decoction on peripheral circulation status of affected limbs and serum thromboxane B 2 (TXB 2) and prostacyclin (PGI 2) levels in patients with traumatic limb fractures after open reduction and internal fixation. Methods:Randomized controlled trial study was conducted. 70 patients with traumatic limb fractures who underwent open reduction and internal fixation in the hospital from February 2020 to February 2023 were set as observation subjects and divided into control group (34 cases) and observation group (36 cases) according to the order of hospitalization. The control group was given conventional method for treatment, and the observation group was given modified Taohong Siwu Decoction adjuvant therapy on the basis of the control group. The treatment for both groups lasted for 2 weeks. TCM symptoms scores were evaluated before and after treatment. Ankle brachial index (ABI) was detected using a Doppler blood flow detector. Color Doppler ultrasound was used to detect the inner diameter, blood flow, and deep veins of the affected limb; serum levels of TXB 2 and PGI 2 were detected by ELISA. Results:After treatment, the total effective rate of clinical efficacy was 91.67% (33/36) in observation group and 73.53% (25/34) in control group, with statistical significance ( P<0.05). After treatment, the observation group had lower scores for qi and blood stasis (2.13 ± 0.43 vs. 3.61 ± 0.96, t=5.85), tendon and bone injury (1.62 ± 0.41 vs. 2.74 ± 0.58, t=9.37), swelling and ecchymosis (1.15 ± 0.31 vs. 2.28 ± 0.52, t=1.12), and liver and kidney deficiency (1.52 ± 0.24 vs. 2.15 ± 0.36, t=8.66) compared to the control group ( P<0.001); after treatment, the ABI (0.83 ± 0.03 vs. 0.74 ± 0.02, t=14.68), vascular diameter [(0.48 ± 0.13) mm vs. (0.42 ± 0.11) mm, t=2.08], and blood flow velocity [(60.24 ± 15.46) cm/s vs. (52.15 ± 12.11) cm/s, t=2.41] in the observation group were higher than those in the control group ( P<0.01 or P<0.05). After treatment, the serum TXB 2 levels [(140.76 ± 16.64) ng/L vs. (168.39 ± 25.28) ng/L, t=5.37] and TXB 2/PGI 2 (6.67 ± 1.24 vs. 9.33 ± 1.69, t=7.54) in the observation group were lower than those in the control group ( P<0.01); the level of PGI 2 [(21.27 ± 2.24) ng/L vs. (18.71 ± 1.79) ng/L, t=5.26] was higher in the control group ( P<0.01). The incidence of deep vein thrombosis after treatment in the observation group was 19.44% (7/36), while in the control group it was 44.12% (15/34), with statistical significance ( χ 2=4.94, P=0.026). Conclusion:Modified Taohong Siwu Decoction adjuvant therapy can enhance the clinical efficacy of patients with traumatic limb fractures after open reduction and internal fixation, relieve the TCM symptoms, improve the peripheral circulation status of affected limbs, reduce the postoperative hypercoagulability and prevent the deep venous thrombosis formation.

Objective To establish a HPLC method and fingerprints study for the simultaneous determination of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin,and ammonium glycyrrhizinatein,and other components in the alcohol extract of Mongolian medicine Qinggan Manu-4.Methods The Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5 μm)was used,with the detection wavelength of 230 nm for paeoniflorin,vitexin and liquiritin,237 nm for loganic acid,250 nm for glycyrrhizic acid,and 270 nm for gentiopicroside.The mobile phase was 0.1%phosphoric acid aqueous solution-acetonitrile with gradient elution.The flow rate was 1.0 mL/min,the column temperature was 30℃,and the injection volume was 10 μL.Results The mass concentrations of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin and ammonium gentiopicroside in the alcohol extract of Mongolian medicine Qinggan Manu-4 showed good linear relationships with the chromatographic peak area in the range of 0.020 3-0.121 0,0.053 3-0.317 7,0.021 3-0.127 0,0.011 5-0.069 0,0.014 1-0.083 8 and 0.035 1-0.209 1 mg/mL,respectively(r≥0.999 8).The average recovery rates of the six components were 94.75%,99.49%,92.32%,95.82%,101.29%and 98.04%,with the RSDs of 1.11%,0.76%,0.99%,2.75%,1.09%and 2.43%,respectively(n=6).The HPLC fingerprint of the alcoholic extract of Mongolian medicine Qinggan Manu-4 was established,using the chromatogram of sample S1 as the reference chromatogram,the control fingerprints were generated through multi-point correction and full-spectrum peak matching.A total of ten common peaks were identified,and after comparison with the reference substance,eight components were identified.Conclusion The established method and fingerprints are accurate,reliable,reproducible and specific,which provide a basis for the quality control and subsequent development of Mongolian medicine Qinggan Manu-4.

Salt signals in tongue are relayed to the nucleus of the solitary tract (NST). This signaling is very important to determine whether to swallow salt-related nutrition or not and suggests some implications in discrimination of salt concentration. Salt concentration-dependent electrical responses in the chorda tympani and the NST were well reported. But salt concentration-dependency and spatial distribution of c-Fos in the NST were not well established. In the present study, NaCl signaling in the NST was studied in urethane-anesthetized rats. The c-Fos immunoreactivity in the six different NST areas along the rostral-caudal axis and six subregions in each of bilateral NST were compared between applications of distilled water and different concentrations of NaCl to the tongue of experimental animals. From this study, salt stimulation with high concentration (1.0 M NaCl) induced significantly higher c-Fos expression in intermediate NST and dorsal-medial and dorsal-middle subregions of the NST compared to distilled water stimulation. The result represents the specific spatial distribution of salt taste perception in the NST.
Animals ; Axis, Cervical Vertebra ; Chorda Tympani Nerve ; Discrimination (Psychology) ; Rats ; Solitary Nucleus ; Taste Perception ; Tongue ; Water

This study was aimed at understanding the genome-wide characterization and variations for three imported novel coronavirus(SARS-CoV-2)strains from different sources in the same period in Jinan at the viral genome level,to provide a sci-entific basis for further improving the prevention and control of COVID-19 outbreaks at Jinan port.We selected nasal and pha-ryngeal swab samples from three cases of imported asymptomatic COVID-19infectionat Jinan port;performed second-genera-tion whole-genome sequencing;and analyzed the variant loci and homology withvarious bioinformatics software.The whole ge-nome sequence of SARS-CoV-2 was successfully obtained from three samples of asymptomatic infected cases,and had full lengths ranging from 29 835 bp to 29 844 bp.Pangolin typing results indicated that the genotypes of the three samples were O-micron BQ.1,BQ.1.1,and BQ.1.12.Compared with the original Wuhan strain,the three samples produced mutations at 77,80,and 78 base sites,respectively,involving 60-63 non-synonymous mutations,mainly in the S and ORF1ab genes.Omicron BQ.1 is an imported variant of the SARS-CoV-2 virus and was detected for the first time at Jinan port.From a molecular biolo-gy perspective,this study provides a theoretical basis for the source tracing and prevention and control of COVID-19 at theport.

In this study,the immunological characteristics of the PhoP protein were explored with a two-component system of Mycobacterium tuberculosis(Mtb).Bioinformatics was used to predict the B and T cell epitopes of the PhoP protein.A re-combinant expression plasmid was constructed by PCR analysis of the phoP sequence and cloning into the prokaryotic expres-sion vector pET-28a(+).Competent Escherichia coli BL21 cells were transformed with the recombinant plasmid and expres-sion was induced with IPTG.The recombinant PhoP protein was purified by affinity chromatography.Serum levels of PhoP-specific antibodies in Mtb-infected mice and tuberculosis(TB)patients were analyzed with an ELISA.BALB/c mice were im-munized with the PhoP recombinant protein by intramuscular injection.Sera of mice were collected and antibody titers were detected with an ELISA and specificity was assessed by West-ern blot analysis.Mouse splenocytes were isolated and the pro-portions of IFN-y-positive cells and cytokine levels were detec-ted with an ELISpot and ELISA,respectively.Bioinformatics i-dentified 24 B cell and 11 T cell epitopes of the PhoP protein.A prokaryotic recombinant vector of PhoP was successfully con-structed and the recombinant PhoP protein was obtained by purification.Specific antibody levels to PhoP in sera of Mtb in-fected mice and TB patients increased significantly,with preci-sion of 99.9％and 82.5％,and specificity of 100％,respectively.PhoP protein immunization successfully induced production of specific antibodies in mice.Stimulated by antigens in vitro,IL-2 and IFN-γ levels were significantly increased in the splenocytes of immunized mice.Immunization with the PhoP protein induce a humoral immune response and Thl-dominated cellular immu-nity,indicating that the PhoP protein was immunogenic with diagnostic efficacy for TB.These results lay a foundation to clari-fy the role of PhoP in Mtb infection and application for diagnosis and prevention of TB.

Objective: To compare the differences in the stress distribution in simulated first molars prepared with three rotary nickel-titanium instruments. Methods : Four simulated first molars were prepared without instruments and with Reciproc, WaveOne and Protaper. Before and after preparation, each simulated molar was scanned by Micro-CT. The data were imported to Mimics 16.0 software to establish three-dimensional models. Finite element analysis was processed with Abaqus 6.14 software under conditions of longitudinal and lateral load. Results: Under vertical load conditions, the maximum von Mises stress of the enamel increased by 1.36%, 21.48% and 20.99% in the Reciproc, WaveOne and Protaper groups, respectively, after preparation, while the maximum von Mises stress of the cementum increased by 55.98%, 41.18% and 33.04%, respectively, and the maximum von Mises stress of the alveolar bone increased by 45.55%, 40.37% and 24.09%, respectively. Under 45° lateral load conditions, the maximum von Mises stress of the enamel increased by 1.79%, -4.58% and 3.82% in the Reciproc, WaveOne and Protaper groups, respectively, after preparation, while the maximum von Mises stress of cementum increased by 16.33%, 7.58% and 4.32%, respectively, and the maximum von Mises stress of the alveolar bone increased by 46.82%, 36.40% and 8.29%, respectively. Under the same conditions, the von Mises stresses of the cementum and alveolar bones of the simulated molars were higher after preparation than before preparation, especially under lateral load conditions. The stress was focused on the border between the crown and the root. The von Mises stress of the cementum and alveolar bones increased much more in the Reciproc group than in the other two groups under both conditions. Conclusion The von Mises stress of simulated molars was greater after preparation than before preparation. The von Mises stress of the cementum and alveolar bones increased much more in the Reciproc group than in the other two groups.