Homo- or heterodimeric compounds that affect dimeric protein function through interaction between monomeric moieties and protein subunits can serve as valuable sources of potent and selective drug candidates. Here, we screened an in-house dimeric natural product collection, and panepocyclinol A (PecA) emerged as a selective and potent STAT3 inhibitor with profound anti-tumor efficacy. Through cross-linking C712/C718 residues in separate STAT3 monomers with two distinct Michael receptors, PecA inhibits STAT3 DNA binding affinity and transcription activity. Molecular dynamics simulation reveals the key conformation changes of STAT3 dimers upon the di-covalent binding with PecA that abolishes its DNA interactions. Furthermore, PecA exhibits high efficacy against anaplastic large T cell lymphoma in vitro and in vivo, especially those with constitutively activated STAT3 or STAT3^Y640F. In summary, our study describes a distinct and effective di-covalent modification for the dimeric compound PecA to disrupt STAT3 function.

ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl₄） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl₄ was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg^-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G₀/G₁ phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G₀/G₁ phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G₀/G₁ phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.

Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

OBJECTIVE: To analyze the clinical phenotype and genetic characteristics of a child with Perlman syndrome. METHODS: Genomic DNA was extracted from peripheral blood samples from the patient and her parents. Whole exome sequencing (WES) was carried out to detect potential variant in the proband. Candidate variant was verified by Sanger sequencing. The pathogenicity of candidate variants was evaluated according to the guidelines of the American College of Medical Genetics and Genomics (ACMG). RESULTS: The results of WES showed that the proband has harbored compound heterozygous variants of the DIS3L2 gene, namely c.2109delC and c.1829.c.1830insC, which were respectively inherited from her mother and father. The results were confirmed by Sanger sequencing. Based on the ACMG guidelines, the two novel variants were both predicted to be pathogenic (PVS1+PS2+PM2). CONCLUSION The compound heterozygous variants of the DIS3L2 gene probably underlay the Perlman syndrome in this patient. Above finding has enriched the spectrum of DIS3L2 gene mutations.
Exoribonucleases ; Female ; Fetal Macrosomia ; Genetic Testing ; Genomics ; Humans ; Mutation ; Whole Exome Sequencing ; Wilms Tumor

Objective:To understand the epidemic characteristics of influenza A (H1N1)pdm09 and hemagglutinin (HA) gene mutations in Hebei province from 2017 to 2020, in order to provide scientific basis for the prevention and control of influenza.Methods:The analysis was conducted on the data of Hebei province, which was down loaded from the Chinese National Influenza Surveillance Network from April 2017 to March 2020, and 37 influenza A (H1N1)pdm09 strains were selected for sequencing and evolutionary analysis of HA gene.Results:A total of 57 670 throat swabs from influenza like illness (ILI) patients in Hebei province were detected, and 8 569 samples were positive for the viral nucleic acid, with the positive rate of 14.86%, influenza A (H1N1)pdm09 accounted for 40.52% (3 472/8 569). The cases were detected throughout the whole 11 regions of Hebei province. All age groups were infected by influenza A (H1N1) virus, and the highest detection rate was in the 25- year age group, the peak detection rate was in winter and spring seasons. The HA genes of 37 strains of influenza A (H1N1) virus belong to 6B.1 branch, and the amino acid homology is from 97.31% to 100.00%. S74R, S164T, S183P and I295V were the major amino acid variation sites in 2017-2018 influenza season. S183P, H126Y, N129D, T185I, L233I and N260D were the major amino acid variation sites of epidemic strains in 2018-2019 influenza season. The variation sites of D187A and Q189E of epidemic strains were found in 2019-2020 influenza season.Conclusions:The prevalence of influenza A(H1N1) pdm09 in Hebei province has an obvious seasonality. The HA gene and its encoded amino acid have been mutating gradually. The strains which were isolated recently were clustered with the 2020-2021 season northern hemisphere vaccine strain A/Guangdong-Maonan/SWL1536/2019.

Objective:To understand the epidemiological characteristics of influenza in Hebei Province from 2018 to 2019, and to analyze the characteristics and variation of hemagglutinin(HA) gene of influenza B-Victoria(BV) strains.Methods:Throat swab specimens of influenza-like cases within 3 days of fever were collected from 28 sentinel hospitals in Hebei province, meanwhile, The surveillance data was collected by the Chinese National Influenza Surveillance Network from April 2018 to March 2019, Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals, and tested by RT-PCR and virus isolation. 14 influenza B-Victoria strains from different regions were selected to sequence HA gene, Phylogenetic tree and the molecular characteristics were analyzed by DNASTAR 7.0 and Mega-X software.Results:From 2018 to 2019, A total of 99 266 cases of influenza-like illness (ILI) were detected from 4 689 103 cases by 28 influenza sentinel hospitals in Hebei Province, the visit percentage of ILI was 2.12%. During the period, 18 730 samples were detected, and 2 752(14.69%) samples were positive tested by RT-PCR, the peak was in the third week of 2019(44.92%), In the early stage of epidemic season, Influenza A(H1N1)pdm09 virus was the main type, while BV virus was the main type in the late stage. HA gene sequence analysis showed that the 14 BV viruses belonged to 162-164 amino acid deletion strains, the amino acid homology between HA sequences was 97.16%-100.00%, and 97.16%-98.95% compared with the vaccine strain B/Colorado/06/2017 recommended by WHO. Compared with the vaccine strains, 14 strains involved 11 amino acid site mutations.Conclusion:Influenza was prevalent in winter and spring in Hebei province from 2018 to 2019, Multiple mutations in antigenic sites of BV viruses might be related to the outbreaks.

Objective:To understand the epidemiological characteristics of influenza in Hebei Province from 2018 to 2019, and to analyze the characteristics and variation of hemagglutinin(HA) gene of influenza B-Victoria(BV) strains.Methods:Throat swab specimens of influenza-like cases within 3 days of fever were collected from 28 sentinel hospitals in Hebei province, meanwhile, The surveillance data was collected by the Chinese National Influenza Surveillance Network from April 2018 to March 2019, Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals, and tested by RT-PCR and virus isolation. 14 influenza B-Victoria strains from different regions were selected to sequence HA gene, Phylogenetic tree and the molecular characteristics were analyzed by DNASTAR 7.0 and Mega-X software.Results:From 2018 to 2019, A total of 99 266 cases of influenza-like illness (ILI) were detected from 4 689 103 cases by 28 influenza sentinel hospitals in Hebei Province, the visit percentage of ILI was 2.12%. During the period, 18 730 samples were detected, and 2 752(14.69%) samples were positive tested by RT-PCR, the peak was in the third week of 2019(44.92%), In the early stage of epidemic season, Influenza A(H1N1)pdm09 virus was the main type, while BV virus was the main type in the late stage. HA gene sequence analysis showed that the 14 BV viruses belonged to 162-164 amino acid deletion strains, the amino acid homology between HA sequences was 97.16%-100.00%, and 97.16%-98.95% compared with the vaccine strain B/Colorado/06/2017 recommended by WHO. Compared with the vaccine strains, 14 strains involved 11 amino acid site mutations.Conclusion:Influenza was prevalent in winter and spring in Hebei province from 2018 to 2019, Multiple mutations in antigenic sites of BV viruses might be related to the outbreaks.

Objective To explore the efficacy of electroacupuncture and massage combined with Yaobi-Zhuyu-Zhitong decoction in the treatment of lumbar disc herniation (LDH). Methods A total of 100 LDH patients with qi stagnation and blood stasis type who met the inclusion criteria were randomly divided into two groups, 50 in each group. The control group was treated with Yaobi-Zhuyu-Zhitong decoction, and the observation group was treated with electroacupuncture and massage on the basis of the control group. Both groups were treated for 4 weeks. The VAS scale was used to evaluate the degree of pain. The Japanese Orthopedic Association Scores (JOA) was used to evaluate the efficacy of low back pain. The Oswestry dysfunction index was used to evaluate the recovery of lumbar function, and the improvement of daily activities of patients after treatment was evaluated.Results After treatment, the VAS score of the observation group was significantly lower than that of the control group, and the JOA score was significantly higher than that of the control group (t value were 9.870, 8.214, P<0.01). The excellent and good rates of Oswestry dysfunction index before and after treatment in the observation group were 34.0％ (17/50) and 86.0％ (43/50), respectively, and the control group was 24.0％ (12/50) and 44.0％(22/50), respectively. The excellent rate of Oswestry dysfunction index in the two groups was significantly higher than that in the same group before treatment (χ2 values were 28.167, 4.456, P<0.01), and the observation group was significantly higher than the control group (χ2=19.385, P<0.001). The scores of daily activities such as walking, weight-bearing, sitting for a long time, bending over, washing, standing, sleeping and turning over in the observation group were significantly higher than those in the control group (t values were 3.689, 1.661, 3.621, 3.621, 3.300, 1.661, 1.461 respectively, all Ps<0.05). The total effective rate was 94.0％ (47/50) and the control group was 76.0％ (38/50). The difference between the two groups was statistically significant (χ2=6.353, P=0.012). Conclusions Electroacupuncture and massage combined with Yaobi-Zhuyu-Zhitong decoction can improve the lumbar function, reduce the degree of lumbar pain, improve the quality of life of the LDH patients with qi stagnation and blood stasis type.

Objective To investigate the application value of single detection and combined detection of 4 kinds of inflammatory indicators of procalcitonin (PCT ) ,high sensitivity C-reactive protein(hs-CRP) ,interleu-kin-6(IL-6) and white blood cell(WBC) in diagnosing type 2 diabetes mellitus(T2DM) bloodstream by analy-zing the levels of peripheral blood PCT ,hs-CRP ,IL-6 and WBC in the T2DM bloodstream infection group and T2DM non-bloodstream infection group .Methods The clinical data in 85 patients with T2DM bloodstream in-fection (T2DM bloodstream infection group ) and contemporaneous 80 cases of T2DM non-bloodstream infec-tion(T2DM non-bloodstream infection group) in this hospital from January 2013 to July 2016 were retrospec-tively analyzed .The levels of various inflammatory indicators in peripheral blood were analyzed .The receiver operating characteristic(ROC) curve of various inflammatory indicators was drawn ,the area under the curve (AUC) and the best cut-off value were calculated .The detection schemes included 24 kinds of schemes such as the single indicator ,2-indicator ,3-indicator and 4-indicator .Results The levels of PCT ,hs-CRP ,IL-6 and WBC in the T2DM bloodstream infection group were significantly higher than those in the T 2DM non-blood-stream infection group ,the difference was statistically significant (P<0 .05) .AUC of PCT ,hs-CRP ,IL-6 and WBC were 0 .909 ,0 .818 ,0 .838 and 0 .760 respectively ,with best cut-off values of 0 .493 ng/mL ,11 .19 ng/mL ,40 .95 pg/mL and 11 .87 × 109/L respectively .The Youden index of PCT was highest (0 .65) and the ac-curacy of IL-6 was highest (83 .33％ ) in the single indicator detection scheme .The Youden index and accuracy of the scheme of PCT/hs-CRP and PCT+hs-CRP+IL-6 were highest in the combined detection scheme .Con-clusion PCT detection has the prominent value in the assisted diagnosis of T 2DM bloodstream infection .Inthe combined detection scheme ,PCT/hs-CRP and PCT+hs-CRP+IL-6 have the highest value in the assisted diagnosis in T2DM bloodstream infection .

Objective To explore the effects of two routes of melatonin (MT) administration including intraperitoneal and caudal vein injection on the behavior,histopathology and the expression of myelin basic protein (MBP) and active caspase-3 protein in focal cerebral ischemic rats.Methods 84 male Sprangue-Dawley rats were randomly divided into normal control group (CON,n=12),middle cerebral artery occlusion group(MCAO,n=24),MT-intraperitoneal group (n=24) and MT-intravenous injection group (n=24) by random number table.Twenty-four hours after ischemia reperfusion (IR),Morris water maze was used to observe the effects of two routes of MT administration on behavior in focal cerebral ischemic rats.7 d after IR,MBP immunohistochemical and hematoxylin eosin (HE) staining were used to examine the expression of MBP in striatum and histopathological changes in hippocampal CA1 region.24 h,72 h and 7 d after IR,the expression of active caspase-3 in hippocampal CA1 region was observed by immunohistochemical staining.Results The average escape latencies in Morris water maze in MT-intravenous injection group at different time points were all lower than those of the MT-intraperitoneal,and they were all lower than those of the MCAO group.Swimming time percentage of target quadrant in MT-intravenous injection group were higher than those of the MT-intraperitoneal,and they were all higher than those of the MCAO group (all P＜0.01);7 d after IR,the results of HE staining showed that the hippocampus cells in MCAO group were disarranged with hyperchromatic nucleus and cytoplasm.More hippocampal cells were observed in MT-intraperitoheal and MT-intravenous injection groups,and they were relatively well arranged.The optical density (OD)of MBP in MT-intravenous injection group (105.60±4.04) was significantly higher than those in MCAO group (95.60±2.07) and MT-intraperitoneal injection group (98.00±4.18) (both P＜0.01).Immunohistochemical results showed that the number of active caspase-3 positive cells in MT-intravenous injection group ((116.93± 12.58)/mm2,(130.16±21.22)/mm2,(88.25±7.80)/mm2) at each time point were significantly lower than those in MT-intraperitoneal injection group ((156.64± 32.54)/mm2,(176.49± 17.44)/mm2,(127.96±16.73)/mm2) (all P＜0.05).At the time points of 24 h and 72 h after IR,there were less active caspase-3 positive cells in MT-intraperitoneal and MT-intravenous injection group compared with those in MCAO group((273.56±32.54)/mm2,(288.63±35.17)/mm2)(all P＜0.01).Conclusion MT administration by both intraperitoneal and intravenous injection can significantly improve the behavior and attenuate the histopathology and white matter damage,and reduce the cell apoptosis in hippocampal CA1 region in focal cerebral ischemic rats,and the therapeutic effects of MT-intravenous injection are better than MT-intraperitoneal injection.