OBJECTIVE To establish the HPLC fingerprint of Xintong granules and the quantitative analysis of multi- components by single-marker method （QAMS） to determine the contents of 7 components， so as to provide a scientific basis for their quality control. METHODS HPLC method was used to establish the fingerprints for 10 batches of Xintong granules （No. S1- S10）， and similarity evaluation， cluster analysis （CA） and partial least squares-discriminant analysis （PLS-DA） were performed. At the same time， the contents of seven components， including puerarin， daidzin， calycosin-7-O- β -D-glucoside， stilbene glycoside， naringin， icariin and tanshinone ⅡA， were determined by QAMS method， and were compared with the results of external standard method. RESULTS A total of 18 common peaks were marked and 7 peaks were identified in the HPLC fingerprints for 10 batches of Xintong granules， namely puerarin （peak 4）， daidzin （peak 7）， calycosin-7-O-β-D-glucoside （peak 9）， stilbene glycoside （peak 10）， naringin （peak 12）， icariin （peak 17）， and tanshinone ⅡA （peak 18）； the similarities among them were more than 0.990， and CA and PLS-DA results showed that S4-S5，S8-S10，S1-S3 and S6-S7 were clustered into three categories， respectively. Using naringin as the internal standard， the contents of puerarin， daidzin， calycosin-7-O-β-D-glucoside， stilbene glycoside， icariin and tanshinone ⅡA were determined to be 7.868 1-10.181 2， 1.709 2-2.374 1， 0.285 2-0.326 3， 1.024 1- 1.523 9， 0.140 2-0.290 4， and 0.077 1-0.219 4 mg/g， respectively， by the QAMS. These results showed no significant differences compared to those obtained by the external standard method. CONCLUSIONS Established HPLC fingerprint and QAMS method are convenient， stable and accurate， which can provide a basis for the quality evaluation of Xintong granules.

This paper summarized Professor ZHOU Zhongying's experience in differentiating and treating hepatitis and liver cirrhosis from deficiency and excess. It is considered that the pathogenesis of hepatitis and liver cirrhosis belongs to deficiency in root and excess in branch， with depletion of liver， spleen and kidney as the root， and constraint and bind of damp-heat and stasis toxin as the branch. Moreover， mutual cause and promotion between deficiency and excess leads to the disease. For general principle of treatment， it is recommended to clear and transform pathogenic excess， supplement deficiency and rectify the healthy qi. In the early stage of hepatitis and cirrhosis， excess pathogen hyperactivity is the main manifestation， which can be treated by clearing and transforming damp-heat and stasis toxin， supplemented by regulating spleen and stomach， with modified Yinchenhao Decoction （茵陈蒿汤） and Biejiajian Pill （鳖甲煎丸）. In the middle and late stages， cases with deficiency-excess complex were more common， which should be treated by clearing damp-heat and stasis toxin， regulating and supplementing liver-spleen-kidney， using medicinals with the function of clearing heat and dispelling damp， dissolving stasis and resolving toxins to treat the branch. Moreover， Liujunzi Decoction （六君子汤）， Yiguan Decoction （一贯煎）plus Erzhi Pill （二至丸） and Buzhong Yiqi Decoction （补中益气汤） modifications are suggested respectively in correspondence to the different kinds of root deficiency including irregular liver and spleen， liver and kidney yin deficiency， and liver-spleen-kidney deficiency.

OBJECTIVE To observe the efficacy of cetrorelix combined with aspirin in preventing early-onset ovarian hyperstimulation syndrome （OHSS）. METHODS A retrospective analysis was conducted on clinical data from 38 early-onset OHSS patients， who received treatment in our hospital from January 1st to July 1st， 2022. These patients were divided into intervention group （19 cases） and control group （19 cases） according to the therapy regimen. On the first day after oocyte retrieval surgery， the control group was given aspirin enteric-coated tablets 100 mg orally until menstruation began. The intervention group was given cetrorelix for injection 0.25 mg subcutaneously， for consecutive 3 days+aspirin enteric-coated tablets （same usage and dosage as the control group）. The first luteal phase， the degree of OHSS， and the ovarian volume， ascites volume， serum estradiol （E2）， white blood cell count （WBC）， hematocrit （HCT）， neutrophil ratio （NEUT%）， D-dimer （DD）， prothrombin time （PT）， fibrinogen （Fib） after oocyte retrieval surgery were observed and measured in 2 groups. RESULTS The first luteal phase was significantly shorter， and the proportions of median and severe OHSS cases were significantly lower in the intervention group compared to the control group （P＜0.05 or P＜0.01）. After oocyte retrieval surgery， the intervention group showed significantly lower ovarian volume， ascites volume， serum E2， WBC， NEUT%， HCT， DD and Fib compared to the control group， but PT of intervention group was signiticantly higher than that of control group （P＜0.05）. CONCLUSIONS Cetrorelix combined with aspirin is more effective in preventing early-onset OHSS than aspirin alone.

Objective To investigate the application value of combining the Demirjian's method with ma-chine learning algorithms for dental age estimation in northern Chinese Han children and adolescents.Methods Oral panoramic images of 10 256 Han individuals aged 5 to 24 years in northern China were collected.The development of eight permanent teeth in the left mandibular was classified into different stages using the Demirjian's method.Various machine learning algorithms,including support vector re-gression(SVR),gradient boosting regression(GBR),linear regression(LR),random forest regression(RFR),and decision tree regression(DTR)were employed.Age estimation models were constructed based on total,female,and male samples respectively using these algorithms.The fitting performance of different machine learning algorithms in these three groups was evaluated.Results SVR demonstrated superior estimation efficiency among all machine learning models in both total and female samples,while GBR showed the best performance in male samples.The mean absolute error(MAE)of the op-timal age estimation model was 1.246 3,1.281 8 and 1.153 8 years in the total,female and male samples,respectively.The optimal age estimation model exhibited varying levels of accuracy across dif-ferent age ranges,which provided relatively accurate age estimations in individuals under 18 years old.Conclusion The machine learning model developed in this study exhibits good age estimation effi-ciency in northern Chinese Han children and adolescents.However,its performance is not ideal when applied to adult population.To improve the accuracy in age estimation,the other variables can be con-sidered.

Objective:To explore significance of eosinophil and eosinophil/lymphocyte ratio(ELR)in predicting allergic asth-ma in children in different seasons.Methods:Retrospective analysis of children with asthma who visited pediatric respiratory depart-ment outpatient clinic and ward of Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 2021 to December 2021,whose allergen specific IgE(sIgE)and peripheral blood cell analysis were complete.sIgE≥0.35 kUA/L were included in allergic asthma group,sIgE<0.35 kUA/L and total allergen IgE<60 kUA/L were included in non-allergic asthma group.General data and changes in peripheral blood cells of two groups were analyzed.Results:There were 1 378 qualified subjeats,including 999(72.5%)in allergic asthma group and 379(27.5%)in non-allergic asthma group.Number of visits in allergic asthma group varied seasonally,with the most in autumn.Peripheral blood lymphocyte count(LYMPH),eosinophil count(EOS)and ELR were all higher in children with allergic asthma than in children with non-allergic asthma(P<0.05),and platelet/lymphocyte ratio(PLR)was lower than that in children with non-allergic asthma(P<0.05).Peripheral blood LYMPH,PLT,EOS and ELR of children with allergic asthma differed between four seasons,which were higher than those of non-allergic asthma in each season in EOS and ELR,LYMPH was significantly higher than that of children with non-allergic asthma in autumn,and PLT was significantly lower than that of children with non-allergic asthma in spring(P<0.05).EOS predicted AUC of spring,summer,autumn and winter were 0.79,0.77,0.71 and 0.64 in children with allergic asthma,and ELR predicted AUC were 0.72,0.48,0.73 and 0.68 in children with allergic asthma.Conclusion:Allergic asthma in children is seasonally variable and peaks in autumn.EOS and ELR in peripheral blood cells in children with allergic asthma are higher than in children with non-allergic asthma in each season of year,LYMPH is significantly higher than children with non-allergic asthma in the fall,and PLT is lower than in children with non-allergic asthma in spring,suggesting that allergic asthma type Ⅱinflammation persists,and EOS and ELR have predictive value for children's allergic asthma.

Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.

Objective: The docking and superantigen activity sites of staphylococcal enterotoxin-like W (SElW) and T cell receptor (TCR) were predicted, and its SElW was cloned, expressed and purified. Methods: AlphaFold was used to predict the 3D structure of SElW protein monomers, and the protein models were evaluated with the help of the SAVES online server from ERRAT, Ramachandran plot, and Verify_3D. The ZDOCK server simulates the docking conformation of SElW and TCR, and the amino acid sequences of SElW and other serotype enterotoxins were aligned. The primers were designed to amplify selw, and the fragment was recombined into the pMD18-T vector and sequenced. Then recombinant plasmid pMD18-T was digested with BamHⅠand Hind Ⅲ. The target fragment was recombined into the expression plasmid pET-28a(+). After identification of the recombinant plasmid, the protein expression was induced by isopropyl-beta-D- thiogalactopyranoside. The SElW expressed in the supernatant was purified by affinity chromatography and quantified by the BCA method. Results: The predicted three-dimensional structure showed that the SElW protein was composed of two domains, the amino-terminal and the carboxy-terminal. The amino-terminal domain was composed of 3 α-helices and 6 β-sheets, and the carboxy-terminal domain included 2 α-helices and 7 antiparallel β-sheets composition. The overall quality factor score of the SElW protein model was 98.08, with 93.24% of the amino acids having a Verify_3D score ≥0.2 and no amino acids located in disallowed regions. The docking conformation with the highest score (1 521.328) was selected as the analysis object, and the 19 hydrogen bonds between the corresponding amino acid residues of SElW and TCR were analyzed by PyMOL. Combined with sequence alignment and the published data, this study predicted and found five important superantigen active sites, namely Y18, N19, W55, C88, and C98. The highly purified soluble recombinant protein SElW was obtained with cloning, expression, and protein purification. Conclusions: The study found five superantigen active sites in SElW protein that need special attention and successfully constructed and expressed the SElW protein, which laid the foundation for further exploration of the immune recognition mechanism of SElW.
Humans ; Enterotoxins/genetics* ; Superantigens/genetics* ; Catalytic Domain ; Selenoprotein W/metabolism* ; Receptors, Antigen, T-Cell

OBJECTIVE: To investigate the anti-coronavirus potential and the corresponding mechanisms of the two ingredients of Reduning Injection: quercetin and luteolin. METHODS: A pseudovirus system was designed to test the efficacy of quercetin and luteolin to inhibit SARS-CoV-2 infection and the corresponding cellular toxicity. Luteolin was tested for its activities against the pseudoviruses of SARS-CoV-2 and its variants. Virtual screening was performed to predict the binding sites by Autodock Vina 1.1.230 and PyMol. To validate docking results, surface plasmon resonance (SPR) was used to measure the binding affinity of the compounds with various proteins of the coronaviruses. Quercetin and luteolin were further tested for their inhibitory effects on other coronaviruses by indirect immunofluorescence assay on rhabdomyosarcoma cells infected with HCoV-OC43. RESULTS: The inhibition of SARS-CoV-2 pseudovirus by luteolin and quercetin were strongly dose-dependent, with concentration for 50% of maximal effect (EC₅₀) of 8.817 and 52.98 µmol/L, respectively. Their cytotoxicity to BHK21-hACE2 were 177.6 and 405.1 µmol/L, respectively. In addition, luetolin significantly blocked the entry of 4 pseudoviruses of SARS-CoV-2 variants, with EC₅₀ lower than 7 µmol/L. Virtual screening and SPR confirmed that luteolin binds to the S-proteins and quercetin binds to the active center of the 3CLpro, PLpro, and helicase proteins. Quercetin and luteolin showed over 99% inhibition against HCoV-OC43. CONCLUSIONS The mechanisms were revealed of quercetin and luteolin inhibiting the infection of SARS-CoV-2 and its variants. Reduning Injection is a promising drug for COVID-19.
Humans ; SARS-CoV-2 ; COVID-19 ; Luteolin ; Quercetin

Radiotherapy is widely used in the management of advanced colorectal cancer (CRC). However, the clinical efficacy is limited by the safe irradiated dose. Sensitizing tumor cells to radiotherapy via interrupting DNA repair is a promising approach to conquering the limitation. The BRCA1-BARD1 complex has been demonstrated to play a critical role in homologous recombination (HR) DSB repair, and its functions may be affected by HERC2 or BAP1. Accumulated evidence illustrates that the ubiquitination-deubiquitination balance is involved in these processes; however, the precise mechanism for the cross-talk among these proteins in HR repair following radiation hasn't been defined. Through activity-based profiling, we identified PT33 as an active entity for HR repair suppression. Subsequently, we revealed that BAP1 serves as a novel molecular target of PT33 via a CRISPR-based deubiquitinase screen. Mechanistically, pharmacological covalent inhibition of BAP1 with PT33 recruits HERC2 to compete with BARD1 for BRCA1 interaction, interrupting HR repair. Consequently, PT33 treatment can substantially enhance the sensitivity of CRC cells to radiotherapy in vitro and in vivo. Overall, these findings provide a mechanistic basis for PT33-induced HR suppression and may guide an effective strategy to improve therapeutic gain.

Paeonia veitchii and P. lactiflora are both original plants of the famous Chinese medicinal drug Paeoniae Radix Rubra in the Chinese Pharmacopoeia. They have important medicinal value and great potential in the flower market. The selection of stable and reliable reference genes is a necessary prerequisite for molecular research on P. veitchii. In this study, two reference genes, Actin and GAPDH, were selected as candidate genes from the transcriptome data of P. veitchii. The expression levels of the two candidate genes in different tissues(phloem, xylem, stem, leaf, petiole, and ovary) and different growth stages(bud stage, flowering stage, and dormant stage) of P. veitchii were detected using real-time fluorescence quantitative technology(qRT-PCR). Then, the stability of the expression of the two reference genes was comprehensively analyzed using geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that the expression patterns of Actin and GAPDH were stable in different tissues and growth stages of P. veitchii. Furthermore, the expression levels of eight genes(Pv-TPS01, Pv-TPS02, Pv-CYP01, Pv-CYP02, Pv-CYP03, Pv-BAHD01, Pv-UGT01, and Pv-UGT02) in different tissues were further detected based on the transcriptome data of P. veitchii. The results showed that when Actin and GAPDH were used as reference genes, the expression trends of the eight genes in different tissues of P. veitchii were consistent, validating the reliability of Actin and GAPDH as reference genes for P. veitchii. In conclusion, this study finds that Actin and GAPDH can be used as reference genes for studying gene expression levels in different tissues and growth stages of P. veitchii.
Real-Time Polymerase Chain Reaction/methods* ; Paeonia/genetics* ; Actins/genetics* ; Reproducibility of Results ; Transcriptome ; Glyceraldehyde-3-Phosphate Dehydrogenases/genetics* ; Reference Standards ; Gene Expression Profiling/methods*