1.Mechanism of Qizhu Kang'ai Prescription for Inhibiting Proliferation of Hepatocellular Carcinoma by Regulating Tumor Metabolic Reprogramming via PCK1/Akt/p21 Signal Axis
Xin ZHONG ; Rui HU ; Jing LI ; Lanfen PENG ; Xingning LIU ; Qi HUANG ; Jialing SUN ; Xinfeng SUN ; Jianping CHEN ; Benqiang CAI ; Xiaozhou ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):26-36
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.
2.Epidemiological analysis of influenza A (H1N1)pdm09 in Hebei province from 2017 to 2020
Guangyue HAN ; Yan LI ; Caixiao JIANG ; Yuyan YANG ; Yanfang LIU ; Lanfen LIU ; Shunxiang QI
Chinese Journal of Experimental and Clinical Virology 2021;35(3):272-275
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.
3.Epidemiological characteristics of influenza and characteristics of hemagglutinin gene of B-Victoria strains in Hebei Province from 2018 to 2019
Guangyue HAN ; Yan LI ; Caixiao JIANG ; Yanfang LIU ; Lanfen LIU ; Shunxiang QI
Chinese Journal of Preventive Medicine 2021;55(7):847-852
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.
4.Epidemiological characteristics of influenza and characteristics of hemagglutinin gene of B-Victoria strains in Hebei Province from 2018 to 2019
Guangyue HAN ; Yan LI ; Caixiao JIANG ; Yanfang LIU ; Lanfen LIU ; Shunxiang QI
Chinese Journal of Preventive Medicine 2021;55(7):847-852
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.
5.Therapeutic effect of massage and electro-acupuncture combined with Yaobi-Zhuyu-Zhitong decoction on lumbar intervertebral disc herniation with qi stagnation and blood stasis type
Sijia FAN ; Chuanyao LIU ; Lanfen HUANG ; Haiwen LIN ; Li XU
International Journal of Traditional Chinese Medicine 2019;41(3):243-247
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.
6.Protective effect of interferon-γ on Chlamydia psittaci acute infection
Guofang TANG ; Lili CHEN ; Liangzhuan LIU ; Chuan WANG ; Lanfen LU ; Yimou WU
Chinese Journal of Zoonoses 2017;33(2):98-103
We investigated the effects of IFN-γ on Chlamydia psittaci (Cps) infection.HeLa cells were treated with different concentrations of recombinant human IFN-γ (5 ng/mL,25 ng/mL,50 ng/mL) after infecting with C.psittaci 6BC,then the number and morphology of C.psittaci inclusion bodies were examined after 48 hours.C57BL/6J mice were intranasally infected with 2 × 106 IFUs C.psittaci 6BC,and intraperitoneally administrated with 10 μg recombinant murine interferon-γ 24 hours prior or post infection,then body weight,activity and survival rate were recorded.The histopathology of mice livers and lungs was analyzed by HE staining on day 5 or day10 post infection.And the chlamydial inclusion bodies were titrated in the lung homogenates of mice sacrificed on day 5 after infection.The inclusion body numbers of recombinant human IFN-γ treated groups (by 5ng/mL,25ng/mL,50ng/mL) were significantly less than that in the control group (23.8±5.1)× 106,(10± 3.58) × 106,(8.0±2.22) × 106,(43.3±11.05)× 106,respectively).And the morphology of inclusion bodies in IFN-γ treated HeLa cells was irregular and much smaller.We also found that IFN-γ could significantly improve the survival rate,reduce acute clinical manifestations and pathological injurery of lung and liver in C.psittaci respiratory tract infected mice model.So we summarized that IFN-γ can mediate strong immunological protection during acute C.psittaci early infection.
7.Viral etiology of acute upper respiratory infection in Hebei Province from 2013 to 2015
Guangyue HAN ; Yan LI ; Yanfang LIU ; Lanfen LIU ; Caixiao JIANG ; Qi LI ; Shunxiang QI
Chinese Journal of Disease Control & Prevention 2017;21(9):891-894
Objective To investigate the viral etiology and the epidemilogy of acute upper respiratory tract infection in Hebei Province from 2013 to 2015,providing scientific basis for diagnosis,prevention and control of the respiratory tract infection.Methods l 551 throat swab samples of the acute upper respiratory infection patients from four hospitals in Hebei Province were collected,Multiplex real-time PCR was used to detect 15 kinds of respiratory tract viruses for all specimens after nucleic acids extraction.Results Totally 714 positive samples were obtained from 1 551 samples,with a positive rate of 46.03%.Human rhinovirus(186,11.99%)was detected as the most common viral species,followed by human parainfluenza 3(167,10.77%),respiratory syncytial virus (122,7.87%),adenovirus (108,6.96%),influenza B virus (56,3.61%),human metapneumovirus(40,2.58%),influenza A virus(39,2.51%),human boca virus (38,2.45%),human parainfluenza 1 (35,2.26%),human coronavirus-229E/NL63 (33,2.13%),human enterovirus (32,2.06%),human parainfluenza 4(31,2.00%),human coronavirus-OC43 (30,1.93%),and human parainfluenza 2(11,0.71%).176 cases (11.35%) were detected as co-infection.The infection rate(56.07%) of under 5 years old group was higher than that of other groups.Conclusions The predominant respiratory viruses,who are responsible for acute upper respiratory tract infections in Hebei Province,are HRV,PIV3,RSV,ADV and IFV.Each virus possess the specific epidemiological feature.
8.Analysis and prenatal diagnosis of PKLR gene mutations in a family with pyruvate kinase deficiency.
Dongliang LI ; Jing ZHANG ; Baoquan JIAO ; Yanli LIU ; Youjun WANG ; Zhiwei WANG ; Wenjing LI ; Lanfen HOU ; Yu SUN ; Hongmou GUO ; Xiao GUO
Chinese Journal of Medical Genetics 2016;33(1):53-56
OBJECTIVETo evaluate the feasibility of genetic and prenatal diagnosis for a family affected with pyruvate kinase deficiency (PKD).
METHODSTargeted sequence capture and high-throughput sequencing technology was used to detect the exons and exon-intron boundaries of the PKLR gene in a clinically suspected PKD patient. Meanwhile, the genotype of the pedigree was validated by Sanger sequencing. Prenatal genetic diagnosis was performed by amniotic fluid sampling after genotype of the mother of the proband was determined.
RESULTSThe proband was found to harbor double heterozygous mutations, c.661G>A (Asp221Asn) and c.1528C>T (Arg510Ter), which resulted in amino acid substitution Asp221Asn and Arg510Ter. Such mutations were confirmed by Sanger sequencing. The mother and father of the proband were detected to have respectively carried c.1528C>T (Arg510Ter) and c.661G>A (Asp221Asn) mutation. The fetus was found to have carried the same mutations as the proband. Following selected abortion, analysis of fetal tissue was consistent with the result of prenatal diagnosis.
CONCLUSIONThe compound mutations of c.661G>A and c.1528C>T of PKLR gene probably underlie the PKD in the family. Prenatal diagnosis of the mutations analysis can facilitate detection of affected fetus in time.
Adult ; Anemia, Hemolytic, Congenital Nonspherocytic ; embryology ; enzymology ; genetics ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Pyruvate Kinase ; deficiency ; genetics ; metabolism ; Pyruvate Metabolism, Inborn Errors ; embryology ; enzymology ; genetics
9.Therapeutic effect of heating and bandage treatment for chronic lymphedema of extremities accompanied with erysipelas: a report of 80 cases.
Ke LI ; Ningfei LIU ; Lanfen FU ; Li WANG ; Jiajia CHEN ; Chen LIANG ; Yixin ZHANG
Chinese Journal of Plastic Surgery 2015;31(1):39-42
OBJECTIVETo investigate the therapeutic effect of heating and bandage treatment for chronic lymphedema of extremities accompanied with erysipelas.
METHODSFrom March 2004 to March 2013, 80 patients with chronic lymphedema of extremities accompanied with erysipelas were analyzed retrospectively. The patients underwent heating treatment (42 degree centigrade) with infrared light machine made by Shanghai Ninth People's Hospital, 2 hours a day, 20 hours for a session. Bandage treatment was adopted after heating treatment. 1 or 2 sessions were performed for each patient every year. The erysipelas occurring frequency, patients subjective feeling, treatment sessions and elastic material usage was recorded during the follow-up period. The erysipelas occurring frequency was tested by the method of rank and inspection. SPSS 17. 0 was used for statistical analysis.
RESULTSAfter heating and bandage treatment, the occurrence frequency of erysipelas was obviously controlled (Z = 7.598, P = 0.000). Erysipelas was not occurred any more in 60 (75%)patients. Remarkable reduction of occurrence frequency of erysipelas caused by various reasons was showed after treatment. Primary and secondary lymphedema after treatment were compared with those before treatment respectively, showing statistical difference (Z = 3.417 and 5.009, P = 0.001 and 0.000). Most of patients felt better subjectively. The relapse rate of erysipelas and lymphedema was lower if keeping using elastic material to give more pressure on extremities after therapy.
CONLUSIONSHeating and bandage treatment can obviously reduce the occurrence frequency of erysipelas. It can improve the quality of patients' lives. Simultaneously, the subsequent elastic material pressure therapy is essential.
Bandages ; Chronic Disease ; Combined Modality Therapy ; methods ; Erysipelas ; complications ; therapy ; Extremities ; Female ; Humans ; Hyperthermia, Induced ; methods ; Lymphedema ; complications ; therapy ; Middle Aged ; Pressure ; Recurrence ; Retrospective Studies ; Time Factors
10.Association of NEDD4 gene polymorphisms with schizophrenia and its clinical characteristics in Chinese Han population.
Xiaojiao BI ; Kaiyan CUI ; Chao HAN ; Mengmeng SUN ; Lina WANG ; Limin YANG ; Dongdong QIAO ; Wenwen HAN ; Lanfen LIU
Chinese Journal of Medical Genetics 2015;32(3):385-390
OBJECTIVETo assess the association of neural precursor cell expressed developmentally down-regulated 4 (NEDD4) with schizophrenia.
METHODSFive single nucleotide polymorphisms (SNPs) of the NEDD4 gene were genotyped by TaqMan SNP genotyping assay in an independent sample of 464 individuals with schizophrenia and 487 healthy controls from eastern Han Chinese population. Clinical data were collected with a general information questionnaire and Positive and Negative Syndrome Scale (PANSS).
RESULTSFrequencies of rs3088077 (allelic: χ2=18.024, P=0.000; genotypic: χ2=16.634, P=0.000), rs7162435 (allelic: χ2=6.771, P=0.009; genotypic: χ2=7.352, P=0.025) and rs2303579 (allelic: χ2=11.253, P=0.001; genotypic: χ2=12.248, P=0.002) were found to be significant different between the two groups. Moreover, TT of rs7162435 was significantly correlated with scores of factors of excitement and hostility (14.53±3.925, F=3.551, P=0.029).
CONCLUSIONrs3088077, rs7162435 and rs2303579 of the NEDD4 gene may be associated with schizophrenia. Moreover, the TT genotype of rs7162435 may increase the severity of excitement and hostility. Our results may provide a clue for delineating the connection between the glutamate hypothesis of schizophrenia and ubiquitination.
Adolescent ; Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Endosomal Sorting Complexes Required for Transport ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Nedd4 Ubiquitin Protein Ligases ; Polymorphism, Single Nucleotide ; Schizophrenia ; enzymology ; ethnology ; genetics ; Ubiquitin-Protein Ligases ; genetics ; Young Adult

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