1.Relationship between CD36 and cell proliferation and migration in hepatocellular carcinoma and its effect on human hepatocellular carcinoma cell xenograft models in nude mice
Lihong ZHANG ; Wei WU ; Guangcai XU ; Peijian ZHANG ; Lide TAO
Cancer Research and Clinic 2024;36(2):98-104
Objective:To observe the expression of CD36 in hepatocellular carcinoma tissues and cell lines, and to investigate the effects of CD36 on the proliferation and migration abilities of human hepatocellular carcinoma cell lines and human hepatocellular carcinoma cell xenograft models in nude mice.Methods:Differences in the expression levels of CD36 transcripts in 371 hepatocellular carcinoma and paracancerous tissues were analyzed based on information from The Cancer Genome Atlas (TCGA) database. Cancer tissues and corresponding paracancerous tissues of 48 hepatocellular carcinoma patients who were diagnosed and underwent surgical treatment at the Affiliated Hospital of Yangzhou University from January 2019 to February 2021 were prospectively collected, and the levels of CD36 mRNA in the tissues were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method. Western blotting was used to detect CD36 protein levels in human hepatocellular carcinoma cell lines Huh7 and HCCLM3 and human normal liver cell line LO2. Plasmids containing CD36 interfering sequences and empty plasmids were transfected into Huh7 cells or HCCLM3 cells for sh-CD36 group and control group, respectively. The CCK-8 assay was used to detect the proliferation ability (expressed as absorbance value) of cells in each group at 0, 12, 24, 36, 48 and 60 h of culture, and the scratch healing assay and Transwell assay were used to detect the migration ability of cells in each group. The Huh7 cells of sh-CD36 group or control group were injected into the axillary subcutis of BALB/c nude mice, with 4 mice in each group, to construct nude mice models of human hepatocellular carcinoma xenografts; the long and short diameters of tumor were measured weekly after 1 week of inoculation, and the tumor volume was calculated. The nude mice were put to death after 5 weeks of inoculation, and the tumor specimens were collected and weighed; the tumor cell morphology was observed under the microscope, and the expressions of CD36 and Ki-67 proteins in the tumor tissues was detected by immunohistochemistry (IHC).Results:Analysis of the data from the TCGA database showed that the level of CD36 transcripts was higher in hepatocellular carcinoma tissues compared with that in paracancerous tissues (4.2±1.8 vs. 3.2±1.5, t = 2.28, P = 0.035). Tissues detection using qRT-PCR in 48 patients with hepatocellular carcinoma showed that the relative expression of CD36 mRNA in hepatocellular carcinoma tissues was higher than that in paracancerous tissues (0.76±0.26 vs. 0.48±0.23, t = 3.52, P < 0.001). Western blotting assay showed that CD36 protein level in Huh7 and HCCLM3 cells was higher than that in LO2 cells, which were (1.42±0.11) times and (1.68±0.16) times higher than LO2 cells, respectively (both P < 0.001). At the mRNA and protein levels, the CD36 of Huh7 and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group (both P < 0.001). CCK-8 assay showed that the proliferative ability of Huh7 cells and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group after 36 and 24 h of culture (both P < 0.01). Scratch healing assay showed that the scratch healing rates of Huh7 cells [(12±3)% vs. (30±5)%, t = 4.01, P < 0.001] and HCCLM3 cells [(15±4)% vs. (29±5)%, t = 4.16, P < 0.001] in the sh-CD36 group were lower than those in the corresponding control group at 48 h of culture; Transwell assay showed that the number of Huh7 cells [(46±6) cells/field of view vs. (88± 6) cells/field of view, t = 5.56, P < 0.001] and HCCLM3 cells [(42±5) cells/field of view vs. (82±7) cells/field of view, t = 5.34, P < 0.001] penetrating into the membrane in 24 h in the sh-CD36 group was less than that in the corresponding control group. Five weeks after subcutaneous injection, the tumor volume [(682±268) mm 3vs. (1 375±512) mm 3, t = 4.73, P = 0.006] and tumor mass [(432±95) mg/mouse vs. (871±109) mg/mouse, t = 6.57, P < 0.001] of nude mice injected with Huh7 cells of the sh-CD36 group were lower than those of nude mice injected with Huh7 cells of the control group; under the microscope, the density of tumor cells in transplanted tumor specimens of nude mice injected with Huh7 cells of the sh-CD36 group was lower than that in nude mice injected with Huh7 cells of the control group, and the expression levels of both CD36 and Ki-67 proteins were also low. Conclusions:CD36 expression is up-regulated in cancer tissues of hepatocellular carcinoma patients and human hepatocellular carcinoma cell lines Huh7 and HCCLM3, and it may associate with cell proliferation and migration of hepatocellular carcinoma. Knockdown of CD36 expression significantly inhibits the proliferation and migration abilities of hepatocellular carcinoma cells in vitro, and inhibits the tumors of human hepatocellular carcinoma cell xenograft models in nude mice.
2.Application of vascularized iliac crest-tensor fascia lata flap in reconstruction of mandibular and oral soft tissue defects
Xiqian WANG ; Guangcai XU ; Liwei PENG ; Chunshi TONG ; Yang WU ; Sichen YAN ; Linlin BU
Chinese Journal of Plastic Surgery 2022;38(7):771-779
Objective:To evaluate the application effect of vascularized iliac crest-tensor fascia lata flap(VIC-TFLF) based on deep circumflex iliac artery and vein in mandibular and oral soft tissue defect reconstruction.Methods:Retrospective analysis of the clinical data of patients with mandible and oral soft tissue defects in Henan Provincial People’s Hospital from October 2020 to March 2022. All cases were performed with computer-aided design and three-dimensional printing to make the models and guide plates. VIC-TFLF was used to repair and reconstruct the oral and mandible defects, the tensor fascia lata flap was used to repair the intraoral soft tissue defect, and the fascia lata was exposed to the mouth directly. After operation, the color, texture and change of intraoral flap were observed, and the recovery and complications were followed up.Results:In this study, 7 patients were included, 4 males and 3 females, aged 27-64 years old, with an average of 50.1 years old. There were 5 cases of squamous cell carcinoma of mandible gingival and cheek, 1 case of postoperative defect of pleomorphic adenocarcinoma of mandible, and 1 case of postoperative defect of ameloblastoma of mandible. According to the defect range of soft and hard tissue after resection, the flap areas of tensor fascia lata were 6.0 cm×3.0 cm-8.0 cm×6.0 cm, and the iliac bones were 3.7 cm×2.4 cm-9.2 cm×2.5 cm. All the composite tissue flaps survived, without distal necrosis, delayed wound healing and marginal fistula. The patients were followed up for 4-19 months, with an average of 11.7 months. The morphology and functions of the mandible and oral soft tissue were well recovered. The surface of tensor fascia lata was exposed to the oral cavity directly, which showed signs of mucosalization within 1 week after operation. The mucosalization was basically completed within 1 month, which was close to normal oral mucosal appearance, and could be reconstructed to produce better oral mucosal appearance in the later period. The wounds in the donor site healed well, and there was no abnormality in lower limb movement and thigh flexion. Among them, 3 patients had numbness on the lateral thigh skin of the donor site 3-5 days after operation. After 6 months of follow-up, the numbness in 2 cases disappeared, and 1 case was significantly reduced.Conclusions:The VIC-TFLF could obtain great appearance and function, with fewer complications and relatively small damage to the donor site, in repair and reconstruction of mandibular defects combined with oral soft tissue defects.
3.Application of vascularized iliac crest-tensor fascia lata flap in reconstruction of mandibular and oral soft tissue defects
Xiqian WANG ; Guangcai XU ; Liwei PENG ; Chunshi TONG ; Yang WU ; Sichen YAN ; Linlin BU
Chinese Journal of Plastic Surgery 2022;38(7):771-779
Objective:To evaluate the application effect of vascularized iliac crest-tensor fascia lata flap(VIC-TFLF) based on deep circumflex iliac artery and vein in mandibular and oral soft tissue defect reconstruction.Methods:Retrospective analysis of the clinical data of patients with mandible and oral soft tissue defects in Henan Provincial People’s Hospital from October 2020 to March 2022. All cases were performed with computer-aided design and three-dimensional printing to make the models and guide plates. VIC-TFLF was used to repair and reconstruct the oral and mandible defects, the tensor fascia lata flap was used to repair the intraoral soft tissue defect, and the fascia lata was exposed to the mouth directly. After operation, the color, texture and change of intraoral flap were observed, and the recovery and complications were followed up.Results:In this study, 7 patients were included, 4 males and 3 females, aged 27-64 years old, with an average of 50.1 years old. There were 5 cases of squamous cell carcinoma of mandible gingival and cheek, 1 case of postoperative defect of pleomorphic adenocarcinoma of mandible, and 1 case of postoperative defect of ameloblastoma of mandible. According to the defect range of soft and hard tissue after resection, the flap areas of tensor fascia lata were 6.0 cm×3.0 cm-8.0 cm×6.0 cm, and the iliac bones were 3.7 cm×2.4 cm-9.2 cm×2.5 cm. All the composite tissue flaps survived, without distal necrosis, delayed wound healing and marginal fistula. The patients were followed up for 4-19 months, with an average of 11.7 months. The morphology and functions of the mandible and oral soft tissue were well recovered. The surface of tensor fascia lata was exposed to the oral cavity directly, which showed signs of mucosalization within 1 week after operation. The mucosalization was basically completed within 1 month, which was close to normal oral mucosal appearance, and could be reconstructed to produce better oral mucosal appearance in the later period. The wounds in the donor site healed well, and there was no abnormality in lower limb movement and thigh flexion. Among them, 3 patients had numbness on the lateral thigh skin of the donor site 3-5 days after operation. After 6 months of follow-up, the numbness in 2 cases disappeared, and 1 case was significantly reduced.Conclusions:The VIC-TFLF could obtain great appearance and function, with fewer complications and relatively small damage to the donor site, in repair and reconstruction of mandibular defects combined with oral soft tissue defects.
4.Association of single nucleotide polymorphisms of miRNA-146a, miRNA-196a2 and miRNA-499 with genetic susceptibility in hepatocellular carcinoma
Lihong ZHANG ; Guangcai XU ; Guodong ZHANG ; Peijian ZHANG ; Lide TAO
Cancer Research and Clinic 2021;33(2):93-98
Objective:To explore the association of miRNA-146a (miR-146a), miRNA-196a2 (miR-196a2), and miRNA-499 (miR-499) single nucleotide polymorphisms with genetic susceptibility in hepatocellular carcinoma.Methods:A case-control study was designed. A total of 175 patients (hepatocellular carcinoma group) in Affiliated Hospital of Yangzhou University from April 2015 to March 2019 and 302 healthy people undergoing physical examination during the same period (the control group) were selected. The genotype distribution of miR-146a, miR-196a2 and miR-499 in the peripheral blood of the two groups were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Logistic regression model was used to analyze the association of 3 genotypes of miRNA, genotypes of hepatitis virus infectors with genetic susceptibility in hepatocellular carcinoma. The relationship between miR-146a gene polymorphism and demography factor as well as the clinical characteristics was also analyzed by using Spearman correlation analysis.Results:In hepatocellular carcinoma group, miR-146a single nucleotide polymorphism of CC, CG, GG site genotypes had 52 (29.7%) cases, 86 (49.1%) cases, 37 (21.1%) cases, respectively; in the control group, the corresponding genotypes had 137 (45.4%) cases, 135 (44.7%) cases and 30 (9.9%) cases, respectively, and the difference in genotype distribution of both groups was statistically significant ( χ2 = 17.23, P < 0.05). There were no statistical differences in genotype distribution of miR-196a2 and miR-499 ( χ2 = 0.51, P = 0.776; χ2 = 0.05, P = 0.976). Single factor logistic regression analysis showed that in the co-dominant model of miR-146a genotype, genotypes of CG ( OR = 1.96, 95% CI 1.13-3.41, P = 0.017) and GG ( OR = 3.30, 95% CI 1.85-5.89, P<0.01) had elevated risk of hepatocellular carcinoma compared with CC genotype. In the dominant model, the risk of hepatocellular carcinoma in CG+GG genotypes was increased compared with that in CC genotype ( OR=1.97, 95% CI 1.33-2.93, P = 0.001); in the recessive model, the risk of hepatocellular carcinoma in GG genotype was increased compared with that in CG+ GG genotype ( OR=2.43, 95% CI 1.44-4.11, P = 0.001). Single factor logistic regression analysis showed that there was no significant difference in the risk of hepatocellular carcinoma in the co-dominant, dominant and recessive models between miR-196a2 and miR-499 genotypes (all P > 0.05). For hepatocellular carcinoma patients with positive hepatitis B virus (HBV), CG genotype had a 2.02-fold (95% CI 1.06-5.07) risk of hepatocellular carcinoma compared with CC genotype, and GG genotype had a 3.12-fold (95% CI 1.66-10.07) risk of hepatocellular carcinoma compared with CC genotype; CG+GG genotype had a 1.91-fold (95% CI 1.85-3.38) compared with CC genotype, GG genotype had a 1.54-fold (95% CI 1.15-6.08) compared with CG+GG genotype. The increasing risk of hepatocellular carcinoma by miR-146a gene polymorphisms was not found in hepatocellular carcinoma patients with hepatitis C virus (HCV) infection or without HBV and HCV infection. Spearman correlation analysis showed that miR-146a gene polymorphisms was not related with age, gender, smoking, drinking, family history of cancer, alanine transaminase and aspartate aminotransferase (all P>0.05). Conclusions:GG and CG genotypes of miR-146a increase the risk of genetic susceptibility in hepatocellular carcinoma, especially for patients with HBV infection. miR-196a2 and miR-499 single nucleotide polymorphisms don't increase the risk of hepatocellular carcinoma.
5. The preliminary analysis on the characteristics of the cluster for the Corona Virus Disease
Haiyan YANG ; Jie XU ; Yan LI ; Xuan LIANG ; Yuefei JIN ; Shuaiyin CHEN ; Rongguang ZHANG ; Weidong ZHANG ; Guangcai DUAN
Chinese Journal of Epidemiology 2020;41(0):623-628
Since December 2019, Corona Virus Disease (COVID-19), a new emerging infection disease occurred in Wuhan, has spread in 27 countries and regions. The clusters of many cases were reported with the epidemic progresses. We collected currently available information for 377 COVID-19 clusters (1 719 cases), excluded the hospital clusters and Hubei cases, during the period from January 1, 2020 to February 20, 2020. There were 297 family clusters (79%), case median 4; 39 clusters of dining (10%), case median 5; 23 clusters of shopping malls or supermarkets (6%), case median 13; 12 clusters of work units (3%), case median 6, and 6 clusters of transportation. We selected 325 cases to estimate the incubation period and found its range is 1 to 20 days, median was 7 days, and mode was 4 days. The analysis of the epidemic situation in a department store in China indicates that there is a possibility of patients as the source of infection during the incubation period of the epidemic. From February 5, 2020 to February 21, 2020, 634 persons were infected in the Diamond Princess Liner. All persons are susceptible to SARS-CoV-2. The older, patients during the incubation period and the worse environment may be the cause of the cases rising. The progress of the two typical outbreaks clearly demonstrates the spread of the early cases in Wuhan. Whatever happens, screening and isolating close contacts remains essential except for clinical treatment during the epidemic. Especially for the healthy people in the epidemic area, isolation is the key.
6.Association between phage-mediated shiga toxin and molecular distribution of CRISPR in Escherichia coli O26 : H11 or NM
Jinzhao LONG ; Yake XU ; Guangcai DUAN ; Wenjuan LIANG ; Huiying LIU ; Shuaiyin CHEN ; Yuanlin XI ; Pengfei WANG ; Yingfang WANG
Chinese Journal of Epidemiology 2017;38(7):944-949
Objective To investigate the association between phage-mediated shiga toxin and molecular distribution of CRISPR in Escherichia (E.) coli O26:H11 or NM.Methods A total of 135 E.coli O26:H11 or NM strains were collected from NCBI database.Software CRT and CRISPR Finder were used to extract CRISPR and Excel was used to assign the spacer of unique number and type CRISPR.And the relationship between CRISPR and stx phage was analyzed.Results All the 135 E.coli O26:H11 or NM strains had the CRISPR.For CRISPRI,CRISPR2.1,CRISPR2.2 and CRISPR3-4,19,22,1 and 1 subtypes were found,respectively.According to the four CRISPR sites,the strains could be divided into 40 subtypes.Stx-phage was only observed in the group C of CRISPR.Compared with E.coli of stx-phage negative,E.coli with stx-phage harbored more spacers.Conclusions CRISPR loci was extensively existed in E.coli O26:H11 or NM,and many subtypes were found in these strains.The presence of stx-phage was related to the molecular distribution of CRISPR in E.coli O26:H11 or NM.CRISPR might be a valuable biomarker to identify strains with high virulent potential.
7.Mechanism research of alleviating rat liver ischemia-reperfusion injury by Astilbin
Yubin XU ; Xu'nan MAO ; Guangcai XU ; Lide TAO ; Jie ZHANG ; Chaochen WANG ; Peijian ZHANG ;
International Journal of Surgery 2016;43(7):450-454,封3
Objective To explore the protective effect of astilbin in hepatic ischemia-reperfusion injury (HIRI).Methods SD rats were divided into Sham group (control group),HIRI group (ischemia-reperfusion group),astilbe (low dose group,middle dose group,high dose group),and estabilished the model of rat HIRI.After liver were reperfused with blood (in 4 h,8 h,16 h),collecting the specimens of blood and liver tissues.Detection of serum alanine aminotransferase (ALT),aspertate aminotransferase (AST);Then observed the changes of liver cell microstructure;Western blot analysised the expression of HMGB1,TLR4,NF-kB,TNF-α in liver tissue.Results The serum ALT levels of Sham group in 4 h,8 h,16 h were (58.11 ±4.81) U/L,(57.12 ± 5.33) U/L,(57.63 ±4.54) U/L,the serum ALT levels of HIRI group in 4 h,8 h,16 h were (540.38 ± 21.41) U/L,(831.21 ± 20.11) U/L,(191.95 ± 15.35) U/L.Compared with Sham group,the serum ALT levels of HIRI group were significantly increased(P < 0.01).Compared with HIRI group,The serum ALT levels of three dose groups in 4 h,8 h,16 h were significantly declined,including high dose group lower the most obvious (The serum ALT levels of high dose group in 4 h,8 h,16 h were (223.75 ± 10.53) U/L,(412.14 ±23.59) U/L,(205.25 ± 15.48) U/L (P <0.01).The results of light microscope indicated that drug groups significantly reduce the liver cell damage.The results of Western blot displayed that High dose group of HMGB1,TLR4 protein expression in 4 h,8 h,16 h drop significantly than HIRI group(P <0.05).High dose group of NFkB,TNF-α protein expression in postoperative 8 h,16 h decrease significantly than HIRI group (P < 0.05),but in postoperative 8 h,there was no statistically significant difference compared with group HIRI (P>0.05).Conclusion Astilbe pretreatment can reduce HIRI and its mechanism may be associated with downregulating the axis of HMGB1/TLR4/NF-kB/TNF-α,proceed to the next inhibiting the inflammatory response.
8.Study on computed tomography features of nasal septum cellule and its clinical significance.
Dingqiang HUANG ; Wanrong LI ; Liming GAO ; Guanqiang XU ; Xiaoyi OU ; Guangcai TANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(5):217-219
OBJECTIVE:
To investigate the features of nasal septum cellule in computed tomographic (CT) images and its clinical significance.
METHOD:
CT scans data of nasal septum in 173 patients were randomly obtained from January 2001 to June 2005. Prevalence and clinical features were summarized in the data of 19 patients with nasal septum cellule retrospectively.
RESULT:
(1) Nineteen cases with nasal septum cellule were found in 173 patients. (2) All nasal septum cellule of 19 cases located in perpendicular plate of the ethmoid bone, in which 8 cases located in upper part of nasal septum and 11 located in middle. (3) There were totally seven patients with nasal diseases related to nasal septum cellule, in which 3 cases with inflammation, 2 cases with bone fracture, 1 case with cholesterol granuloma, 1 case with mucocele.
CONCLUSION
Nasal septum cellule is an anatomic variation of nasal septum bone, and its features can provide further understanding of some diseases related to nasal septum cellule.
Adolescent
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Adult
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Aged
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Ethmoid Bone
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diagnostic imaging
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Female
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Humans
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Male
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Middle Aged
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Nasal Bone
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diagnostic imaging
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Nasal Septum
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diagnostic imaging
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Retrospective Studies
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Tomography, X-Ray Computed
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Young Adult
9.Advance in Research of Sub-health(review)
Xianglan LI ; Guangcai ZHAO ; Yongling XU ; Tong ZHOU
Chinese Journal of Rehabilitation Theory and Practice 2008;14(10):942-946
To improve human cognition of sub-health condition by analyzing current situation of sub-health researches,the authors of this article reviewed the concept of sub-health,etiological factor,epidemiology,diagnostic code and other sides.
10.Correlation between body mass index and physical qualities and functions in 2 056 adults from Guangzhou
Guangcai ZHAO ; Weilong HE ; Tong ZHOU ; Yongling XU
Chinese Journal of Tissue Engineering Research 2007;0(15):-
AIM: The auxology-related indexes are important for tissue construction. In this article, we investigated the incidence of overweight and obesity and their effects on physical functions and qualities of Guangzhou adults in 2005. METHODS: 2 056 adults including 1 027 males and 1 029 females were randomly selected from Guangzhou in 2005 for physical examination. The anthropometric data such as body mass and height were collected to calculate body mass index (BMI). The subjects were divided according to the BMI criteria recommend by Group of China Obesity Task Force, and their physical functions and qualities were analyzed. RESULTS: In 2005, the detection rate of overweight and obesity in adult males were 28.7% and 5.2%, and in females were 18.6% and 5.3%. The values of diastole pressure and systolic pressure were increased with BMI, but the change tendency of step index and vital capacity/body mass was decreased with BMI. The rate of hypertension was increased with BMI, and male subjects had higher rate of hypertension than that of female subjects in all groups. Obese adult had the poorest values in vertical jump and sit-and-reach test. Those people with normal BMI had better balance ability than that of overweight and obese people. CONCLUSION: The rates of overweight and obesity in Guangzhou adult show a high level. The physical functions and qualities may be improved when the body mass increases or decreases in a certain range, but obesity has impact to those values.


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