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.Correlation between irregular antibody profiles of non-Rh blood group system and incidence of hemolytic disease of the newborn, 53 cases
Jing QIAO ; Qingsheng LIU ; Xinfeng PANG ; Ruiguang GAO ; Buqing GUO ; Rencun WANG ; Xin ZHANG ; Ailing WANG ; Lingli BI
Chinese Journal of Blood Transfusion 2021;34(8):874-877
【Objective】 To retrospectively analyze the irregular antibodies in 6 blood group systems other than the Rh blood group system in 53 pregnant women and analyze its correlation with the occurrence of hemolytic disease of the newborn(HDN). 【Methods】 19 473 pregnant women were screened for irregular antibodies by microgel detection technology combined with anti-human globulin (IgG+ C3d), and the positive samples screened out were further confirmed to understand the types and titers of irregular antibodies. Irregular antibody type determination experiment: IgG type irregular antibody titer was determined after mercaptoethanol (2-Me) inactivated the serum of the irregular antibody positive specimen, and then IgG and IgM type were determined by comparing the titer levels of irregular antibody. Three hemolysis tests and total bilirubin tests were performed on umbilical cord blood during delivery to analyze the level of jaundice and the occurrence of HDN. 【Results】 53 cases of irregular antibodies other than the Rh blood group system were detected in 19 473 pregnant women, with a positive rate of 0.27%, mainly MNS and Lewis blood group system.The incidence of HDN was 39.6% (21/53). There were 27 cases of IgM, 7 IgG, and 19 IgM + IgG. Comparison of total bilirubin detection between the low titer group (≤8) and the high titer group (>8) : the latter was significantly higher than the former (P<0.05); IgG antibody subtypes: IgG1 of the latter significantly increased (P<0.05), and so was IgG3 in former (P<0.05). There was a significant positive correlation between IgG1, IgG3 and total bilirubin. The area under the curve of IgG1+ IgG3 for HDN diagnosis, the sensitivity and specificity were 0.953, 0.900, and 0.967, respectively. 【Conclusion】 Other than Rh blood group system, irregular antibodies are mainly distributed in MNS and Lewis blood group system. The incidence of HDN is higher in Kell, Duffy and Kidd blood group systems after producing irregular antibodies. Non-antibody types are mostly IgM type or IgM + IgG mixed, and the incidence of HDN is not high; Patients with poor maternal history, either high or low titer, can be classified into IgG1 and IgG3 in early stages, and those with Abnormal results should be included into the perinatal management of high-risk women with regular checking.
3.Thromboelastogram parameters and MTHFR gene polymorphism in the diagnosis of adverse fetal pregnancy outcomes
Jing QIAO ; Xinfeng PANG ; Rencun WANG ; Xin ZHANG ; Ailing WANG
Chinese Journal of Blood Transfusion 2021;34(7):728-731
【Objective】 To retrospectively analyze the situation of patients with adverse fetal outcomes by thromboelastogram (TEG) parameters and, MTHFR gene polymorphism, so as to provide molecular biological diagnosis basis for patients with adverse pregnancy outcomes, and a new scheme for early prevention and treatment of women of childbearing age with MTHFR gene polymorphism. 【Methods】 A total of 100 women with adverse fetal pregnancy outcomes were selected as the adverse pregnancy group, and 100 healthy women of childbearing age with normal pregnancy history were selected as the controls. MTHFR gene C677T and A1298C polymorphisms were detected by polymerase chain reaction (PCR). TEG and blood coagulation were detected in the experimental group. 【Results】 The A1298C gene polymorphism(AA、CC、AC; A、C) was similar in both adverse pregnancy group and the controls. The frequency distribution of C, T allele of MTHFR gene C677T was statistically significant (χ2=4.60, P<0.05, OR =1.645, 95% CI: 1.042~2.595). TT and CT+ CC types showed significant different association with the factors of stillbirth(χ2 =7.49, P<0.05). MA value of TEG in the diagnosis of TT type of C677T genotypes MTHFR in 32 patients with adverse pregnancy outcome was analyzed. The area under the AUC curve of MA value was 0.795. 【Conclusion】 MTHFR C677T polymorphism TT with TEG parameter hypercoagulability is an important risk factor in the occurrence of pregnancy stillbirth in adverse pregnancy outcomes.
4.Application value of DSC-PWI in ischemic cerebrovascular disease
Yanping JING ; Bin LUO ; Zhengrong GAO ; Xinfeng XU ; Lidong YAO ; Tao CHENG ; Yan ZHANG ; Jingliang CHENG
Journal of Chinese Physician 2020;22(3):369-373,378
Objective:To explore the application value in cerebral blood perfusion status of dynamic susceptibility contrast enhanced perfusion weighted imaging (DSC-PWI) in ischemic cerebrovascular disease (ICVD).Methods:Retrospective analysis of 31 cases ICVD head magnetic resonance imaging (MRI), diffusion weighted imaging (DWI), magnetic resonance angiography (MRA) and DSC-PWI image in clinical diagnosis, and selectively analyze parameters including relative cerebral blood flow (rCBF), relative cerebral blood volume (rCBV), mean transit time (MTT) and time to peak (TTP) for generalize its characteristics.Results:31 cases of ICVD, 16 cases hypoperfusion, rCBF decreased significantly, rCBV decreased, MTT and TTP prolonged, include 7 cases of large vessel disease and 5 cases of small vessel disease. The collateral circulation formed 9 cases, including 3 cases with good compensation, rCBF normal, rCBV normal, MTT and TTP prolonged, 6 cases with bad compensation, rCBF decreased, rCBV normal or increased, MTT and TTP prolonged. The blood reperfusion in 3 cases, rCBF normal or slightly increased, rCBV increased, MTT shortened or normal, and TTP shortened. The excessive perfusion in 3 cases, rCBF significantly increased and rCBV significantly increased, with MTT and TTP shortened.Conclusions:DSC-PWI can reliably reflect the perfusion state and collateral circulation compensation of ICVD, so as to guide the selection of clinical treatment program and significantly improve the prognosis of patients.
5.Effect of acellular repair patch of small intestinal submucosa of porcine in repairing of soft tissue defects of hand
Chengwu ZANG ; Fanliang ZHANG ; Cheng YANG ; Yongxiang CHEN ; Wenzhi ZHANG ; Xinfeng JING ; Ji MA ; Rui CONG
Chinese Journal of Microsurgery 2020;43(2):157-160
Objective:To compair the outcomes of repairing soft tissue defects of hand between a biodegrad- able repair patch—porcine small intestinal submucosa (SIS) and skin grafting.Methods:From December, 2017 to December, 2018, 36 cases of hand soft tissue defect were treated and analyzed retrospectively. According to the defect area and treatment methods, 36 cases were divided into 2 groups: SIS group (21 cases) and grafting group(15 cases). In SIS group, the area of soft tissue defects was 2.0 cm×1.5 cm-9.0 cm×3.5 cm with an average of 5.3 cm×2.1 cm, treat- ed with SIS; In grafting group, the area of soft tissue defects was 9.0 cm×4.0 cm-16.0 cm×9.0 cm with an average of 12.0 cm×8.5 cm, treated with autologous skin grafting after wet dressing. Wound healing was evaluated at 14, 21 and 28 days, and 3 months after the surgery according to the appearance of colour, elasticity, sensory recovery and prog- noses of partial tendon exposure.Results:All patients were followed-up for 3 to 10 months, with an average of 5 months. All wounds in both groups were completely healed; the appearance was normal, and the skin elasticity and sensation had recovered. Sensation recovery in SIS group: 14 cases were good (66.6%), 5 cases were fine (23.8%), and 2 cases were bad (9.6%); in grafting group: 9 cases were good (60.0%), 4 cases were fine (26.0%), and 2 cases were bad (14.0% ). Wound healing effect in SIS group: 14 cases were good, 5 cases were fine, and 2 cases were bad; in grafting group: 9 cases were good, 4 cases were fine, and 2 cases were bad.Conclusion:The SIS patch can be used in the reconstruction of soft tissue defects in hand. There was no significant difference in colour compared to the sur- rounding skin and left no scar. The patch is an ideal repair material for superficial skin defects.
6.Characteristics of impulsiveness and aggressiveness in patients with hepatolenticular degeneration and its relationship with brain structure
Ping JIN ; Gongqiang WANG ; Jing YU ; Xinfeng MA ; Song WANG ; Yongzhu HAN ; Renmin YANG
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(2):133-137
Objective To investigate the impulsivity and aggressiveness characteristics of patients with hepatolenticular degeneration (HLD) and its relationship with brain structure.Methods The Chinese version of the Barratt impulsiveness scale,11 version (BIS-1 1-C) and Buss-Perry aggression questionnaire (BPAQ) were assessed in 78 patients with hepatolenticular degeneration(HLD group) and 86 normal adults (health control group).HLD patients were examined by 3.0T magnetic resonance imaging (MRI).The differences in impulsivity and aggressiveness of the two groups were compared,and the relationship was analyzed between impulsivity,aggressiveness and different brain structures in patients with HLD.Results The total impulsive score,unplanned factor score,the total aggressive score and anger factor score of patients with cerebral HLD (61.74±9.82,26.08±5.06,82.71 ± 15.92,20.06± 5.74,respectively) were higher than those in patients with hepatic HLD (56.73±7.11,23.02±4.20,72.84± 11.15,16.64±5.01,respectively),and health control group(52.19±7.53,21.50± 3.93,64.64±9.83,14.27 ±4.38,respectively),and the differences were significant (F=3.193,4.646,11.830,8.270,all P<0.05).Total impulsive score was positively correlated with aggressive score and physical aggression in HLD group(r=0.299,0.290,both P<0.05).Unplanned score was positively correlated with aggressive total score (r=0.324) and physical aggression (r=0.320) in HLD group (P<0.05).Frontal lobe injury was the influencing factor of total impulsive score(B=10.263,95%CI=0.467-19.946,P=0.008),attention score(B=2.837,95%CI=0.382-5.600,P=0.010) and unplanned factor score (B=3.977,95% CI=0.848-8.502,P=0.046).Thalamus injury was the influencing factor of aggressive total score and its factor score.Caudate nucleus injury was the influencing factor of aggressive total score (B=10.030,95% CI=3.351-18.039,P=0.017) and physical aggression score (B =4.432,95% CI=1.193-7.729,P=0.016).Conclusion Patients with HLD have higher impulsive and aggressive tendencies,which are mainly manifested in unplanned impulsive and anger tendencies.Brain injury may be an important factor affecting impulsiveness and aggression in patients with HLD.Impulsiveness is related with frontal lobe injury and aggression to thalamus and caudate nucleus injury.Impulsiveness and aggressiveness in patients with HLD are not caused by damage to isolated brain areas,but are related to damage to multiple brain areas.
7.Technical process and quality control of precise thoracoscopic lung segmentectomy
CHEN Liang ; WANG Jun ; WU Weibing ; ZHU Quan ; WEN Wei, ; XU Jing ; HE Zhicheng ; XU Xinfeng ; PAN Xianglong
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(1):21-28
Thoracoscopic pulmonary segmentectomy is one of the important treatment methods for early lung cancer. Only the premise of surgical precision can make the radical resection of lesions and maximum reservation of healthy lung tissue be simultaneously guaranteed. As a representative of the precise lung operation, the “cone-shaped lung segmentectomy” focuses on the lesion, and combines the anatomical characteristic of the patient to design individualized operation scheme. The technological core consists of three parts, three dimensional-computed tomography bronchography and angiography (3D-CTBA) surgery path planning, accurate definition of intersegmental demarcation and anatomic dissection of intersegmental borders along the demarcation. This paper aims to explore the technical process and quality control of the key techniques of thoracoscopic precise segmentectomy, so as to standardize the segmentectomy procedure under the principle of radical and minimally invasive therapy.
8.Clinical application of the lateral branch of lateral circumflex femoral artery in the anterolateral thigh perforator flap
Chengwu ZANG ; Rui CONG ; Wenzhi ZHANG ; Chao LIANG ; Xinfeng JING ; Ji MA ; Yongxiang CHEN ; Yule ZHU
Chinese Journal of Microsurgery 2019;42(3):213-217
Objective To investigate the feasibility and technique of using the anterolateral thigh perforator flap pedicled with the lateral branch of the lateral circumflex femoral artery (LBLCFA) to repair soft tissue defect of extremities.Methods Eighty-six cases of anterolateral thigh perforator flap transplantation were performed from May,2014 to May,2018.A total of 37 cases of soft tissue defect of extremities were treated by anterolateral thigh perforator flap used the LBLCFA as vascular pedicle,of which there were 19 cases of upper limbs and 18 of lower limbs.There were 27 cases of defects caused by trauma,and 10 by soft tissue tumor resection.The flaps were designed centering around the point proximal to the midpoint of the iliac-patellar line.The dimensions of soft tissue defect were from 9.0 cm×6.0 cm to 26.0 cm×10.0 cm,and the flap were from 10.0 cm×7.0 cm to 27.0 cm×11.0 cm.The length of vascular pedicle ranged of 7.0-13.0 cm,with an average of 11.5 cm.The donor sites were directly sutured.All of the patients were followed-up regularly in the outpatient department.Results All the flaps survived and the donor sites were primarily healed.Of these 37 cases,2 trauma patients and 2 patients treated with local radiotherapy had poor wound healing,but still healed after multiple dressing changes.All the patients were followed-up for 3-26 months,with an average of 13 months.The texture,color and elasticity of the flap were similar to the surrounding tissue of the recipient sites,while only a linear scar remained at the donor sites.Ten tumor patients were treated with routine radiotherapy and chemotherapy after the repairation;there was no tumor recurrence during the follow-up period.Conclusion The LBLCFA gives off a relatively large and thick perforator proximal 5.0-7.0 cm of the iliacpatellar line,which locates in the upper lateral side,travels parallel to the vastus lateralis,and give off the skin and muscular perforators.The lateral branch can be used as pedicle to make into perforator flap or chimeric flap,which provides a novel selection of vascular pedicel for anterolateral thigh perforator flap.When the descending branch does not provide a thick and large perforator,the LBLCFA has important practical value and is worth utilizing in the clinic.
9.Learning curve of totally thoracoscopic pulmonary segmentectomy.
Weibing WU ; Jing XU ; Wei WEN ; Yue YU ; Xinfeng XU ; Quan ZHU ; Liang CHEN
Frontiers of Medicine 2018;12(5):586-592
Totally thoracoscopic pulmonary segmentectomy (TTPS) is a feasible and safe technique that requires advanced thoracoscopic skills and knowledge of pulmonary anatomy. However, data describing the learning curve of TTPS have yet to be obtained. In this study, 128 patients who underwent TTPS between September 2010 and December 2013 were retrospectively analyzed to evaluate the learning curve and were divided chronologically into three phases, namely, ascending phase (A), plateau phase (B), and descending phase (C), through cumulative summation (CUSUM) for operative time (OT). Phases A, B, and C comprised 39, 33, and 56 cases, respectively. OT and blood loss decreased significantly from phases A to C (P < 0.01), and the frequency of intraoperative bronchoscopy for target bronchus identification decreased gradually (A, 8/39; B, 4/33; C, 3/56; P = 0.06). No significant differences were observed in demographic factors, conversion, complications, hospital stay, and retrieved lymph nodes among the three phases. Surgical outcomes and techniques improved with experience and volume. CUSUMOT indicated that the learning curve of TTPS should be more than 72 cases.
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10.Isolation, antimicrobial resistance profile and virulence gene analysis of Campylobacter spp.originated from duck
Hang ZENG ; Junfeng PENG ; Jing HUANG ; Weiwei YAN ; Peng CHEN ; Kang ZHOU ; Likou ZOU ; Yong HUANG ; Xinfeng HAN ; Shuliang LIU
Chinese Journal of Zoonoses 2017;33(1):15-21
To investigate the contamination conditions of Campylobacter spp.in duck production chain and its antimicrobial resistance,virulence gene distribution,samples were collected at the duck slaughterhouse according to GB 4789.9-2014.Triplex PCR assay was applied to identify the species of Campylobacter and the recovered Campylobacter strains were tested for the antimicrobial susceptibility against 8 kinds of antimicrobial agents using a broth microdilution method,the susceptibility results were determined according to the NARMS criteria (2011).Subsequently,4 virulence genes were detected by PCR method.The result showed that 187 Campylobacter isolates were obtained from 489 samples,including 160 C.jejuni,130 C.coli and 10 unidentified Campylobacter isolates.The total isolation rate of Campylobacter was 38.24%.The prevalence of Camnpytobacter before slaughtering,at depilation stage,evisceration stage and duck products was 76.33%,5.62%,24.00%,and 0%,respectively.The Campylobacter isolates were most frequently resistant to tetracycline (95.72 %),followed by resistance to clindamycin(90.91%),the resistance rate of azithromycin (63.64%) was in the middle,the resistance rates of ciprofloxacin(31.02%),gentamicin(34.76%),nalidixic acid (37.43 %),erythromycin (41.18 %),chloramphenicol (41.18%) were relatively low.The multi-drug resistance was common among Campylobacter isolates with a rate of 72.19%.The prevalence of adhesion-associated gene cadF,flagellin gene flaA,invasion associated protein gene iamA,toxin regulation gene cdtB was 100%,80.75 %,71.12% and 94.65%,respectively.The results indicated that the Campylobacter contamination occurred in the slaughtering procedures of duck,and the antimicrobial resistance of Campylobacter isolates was relatively serious.In addition,the virulence-associated genes were detected widely among Campylobacter isolates.Therefore,the supervision of antimicrobial agents using at rearing stage should be strengthened,along with health management in duck production chain.


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