1.Progress of proteomics in screening the diagnosis marker of pancreatic cancer
International Journal of Surgery 2008;35(5):321-324
The proteome is the entire complement of proteins expressed by a genome, cell, tissue or organism,which are all types of proteins expressed by one cell in the specific physiological or pathological states. The study of proteomics not only can help us to understand the general rules of activity of proteins,but also illuminate the pathologic mechanisms of numerous diseases. By proteomic analysis of normal individual and pathological individuals, we can find some disease-specific protein molecules, which not only can be a new target for drug design, but also can be a molecular markers of some diseases especially tumors.
2.The expressions and clinical significance of complements 3, 4B1 and apolipoprotein E in pancreatic cancer
Jiong CHEN ; Chunsheng ZHENG ; Houkuo TANG
Chinese Journal of Digestion 2011;31(7):442-445
Objective To investigate the expressions of complements 3 (C3), 4B1 (C4B1) and apolipoprotein E (ApoE) in pancreatic cancer and relations with TNM staging and lymph node metastasis of pancreatic cancer. Methods Thirty-eight pancreatic cancer biopsy specimens, 20 fresh pancreatic cancer specimens and 20 adjacent normal tissues of pancreatic cancer were collected. The expressions of C3, C4B1, ApoE in pancreatic cancers and normal pancreatic tissues were detected by immunohistochemistry and Western-Blot, the positive expression rates of C3, C4B1, ApoE and the differences in gray scale were also observed. Their association with pancreatic cancer TNM staging and lymph node metastasis were analyzed by SPSS 13.0. Results The expression rates of C3, C4B1, ApoE in pancreatic cancer were 73.68% (28/38), 86.84%(29/38) and 76.31% (33/38) respectively, higher than those in normal pancreatic tissues which were 42.11% (16/38), 26.32% (10/38) and 42.11% (16/38) accordingly, the differences were statistically significant (χ2 was 7.77, 19.01, 16.6, and P value were 0.01, 0.00, 0.00 respectively). The gray scale of C3, C4B1 and ApoE in pancreatic cancer were 1.63±0.28,1.25±0.18 and 2.57±0.22 respectively, higher than those in normal pancreatic tissue (0.88±0.19,0.65±0.13,1.28±0.24 respectively), the differences were statistically significant (t value were 9.93,11.81,17.71 and all P value were 0.00, respectively). There was no association between C3 and TNM staging or lymphatic metastasis of pancreatic cancer. C4B1 and ApoE were closely related with TNM stage and lymph node metastases. The expressions of C4B1 and ApoE in stage Ⅱ to Ⅳ pancreatic cancer or with lymphatic metastasis were significantly higher than those in stage Ⅰpancreatic cancer and those without lymph node metastasis. Conclusion C3, C4B1 and ApoE were all highly expressed in pancreatic cancer. C3 was only involved in early event in pancreatic cancer, not related with development of pancreatic cancer. C4B1 and ApoE were involved in tumor growth and metastasis.
3.Apolipoprotein E and complement C4b1 expressions and its significance in human pancreatic carcinoma
Jiong CHEN ; Chunsheng ZHENG ; Houkuo TANG
Chinese Journal of Pancreatology 2011;11(2):89-91
Objective To investigate the expression of apolipoprotein E (ApoE) and complement C4b1 in pancreatic carcinoma and study its significance. Methods Immunohistochemistry was used to detect the expression of ApoE and C4b1 protein in 38 cases of pancreatic carcinoma tissues and adjacent normal pancreatic tissues, and RT-PCR was used to detect the expression of ApoE and C4b1 mRNA in 20 cases of pancreatic carcinoma tissues and adjancent normal pancreatic tissues. The relevance of ApoE and C4b1 expressions to the biological features of pancreatic carcinoma were analyzed. Results The positive rates of ApoE and C4b1 expressions are 86.8% (33/38) and 76.3% (29/38) in pancreatic carcinoma, respectively,which were significantly higher than those in normal pancreatic tissues [42.1% (16/38) and 26.3% ( 10/38 ),P < 0.01]. The positive rates of ApoE and C4b1 expressions [78.3% ( 18/23 ) and 73.9% ( 17/23 )] in patients with metastasis were significantly higher than in those without metastasis [(33.3% (5/15) and 40.0%(6/15), P < 0.05). Significantly higher expressions of ApoE and C4b1 mRNA were noted in pancreatic carcinoma(4.83 ± 0.65 and 7.94 ± 0. 95 ) than those in the normal pancreatic tissue ( 1.78 ± 0.74and 1.22 ±0.57, P < 0.01 ), and patients with metastasis showed significantly higher expression of ApoE and C4b1 mRNA (5.05 ±0.71 and 8.24 ± 1.07) than those without metastasis (4.42 ±0.25 and 7.39 ±0.15,P < 0.05). Conclusions ApoE and C4b1 were highly expressed in pancreatic cancer, and may be closely related with lymph node metastasis.
5.C3 and ApoE expressions and their significance in human pancreatic carcinoma
Chunsheng ZHENG ; Jiong CHEN ; Houkuo TANG
International Journal of Surgery 2011;38(3):154-157,封3
Objective To investigate the association of C3 and ApoE expressions with lymph node metastasis and clinical pathological stage of human pancreatic cancer. Methods Immunohistochemistry was used to detect the expression of C3 and ApoE in pancreatic cancer's tissues and normal pancreatic tissues,and the relevance of C3 and APoE expressions to the metastasis of pancreatic cancer was analyzed. Results The positive rates of C3 and ApoE expressions were 73.68% and 86.84% in pancreatic cancer,significantly higher than those in normal pancreas tissues (42.11% and 42.11%, P<0.01 ). The positive rate of C3 expressions in pancreatic cancer of lymph node metastasis was 56.52%, in those without lymph node metastasis was 46.67% (P=0.741). The positive rate of C3 expressions in pancreatic cancer of stage Ⅰ was 57.14%, in those of stage Ⅱ - Ⅳ was 77.42% (P=0.194). The positive rate of ApoE expressions in pancreatic cancer of lymph node metastasis was 78.26%, significantly higher than those without lymph node metastasis (33. 33%, P<0.01). The positive rate of ApoE expressions in pancreatic cancer of stage Ⅰ was 57. 14%, significantly lower than those of stage Ⅱ - Ⅳ (93.55%, P <0.05 ). Conclusions C3 and ApoEare all overexpressed in pancreatic cancer. C3 is not related with tumor's lymph node metastasis and clinical stages, may be marker for early diagnosis of pancreatic cancer. ApoE is closely related with tumor' s development, may reflect the biological behavior of pancreatic cancer.
6.Effect of 5-Aza-dC on the methylation and expression of TFPI-2 gene In pancreatic cancer PANCI cell line
Zhigang TANG ; Hao ZHENG ; Qiang HUANG ; Jiong CHEN
Chinese Journal of Pancreatology 2012;12(2):92-94
ObjectivesTo investigate the effects of 5-aza-2-deoxycytidine(5-Aza-dC),a methylation inhibitor,on the expression and methylation of tissue factor pathway Inhibitor (TFPI-2) gene in PANC1 cell line of pancreatic cancer.MethodsPANC1 cell lines were treated with different dosages of 5-Aza-dC ( 1× 10-7,5 × 10-7,1× 10 -6 mol/L).The status of TFPI-2 methylafion and expressions of TFPI-2 mRNA and protein were determined by MSP,RT-PCR,and Western blot.Results TFPI-2 gene CpG island was completely methylated,and there was no expression of TFPI-2 mRNA and protein without 5-Aza-dC treatment.After treatment with different dosages of 5-Aza-dC( 1 × 10-7,5 × 10 -7,1 × 10-6 mol/L),TFPI-2 gene CpG hypermethylation was reversed from incomplete methylated to complete non-methylated.The relative expressions of TFPI-2 mRNA were 0.211± 0.087,O.327 ± 0.068,0.609 ± 0.017; and the relative expressions of TFPI-2 protein were O.429 ± O.121,O.675 ± O.044,1.132 ± O.124 in a dose-dependent manner ( P <0.05 ).ConclusionsThe hypermethylatien of promoter region may be the primary reason for TFPI-2 gene expression down-rogtdation and inactivation.5-Aza-dC may reverse the hypermethylation of TFPI-2 gene,and induce the m-expression of TFPI-2 mRNA and protein.
7.Application of urinary albuminuria at different time in the monitoring of early impairment in kidney of diabetes mellitus
Wei GUO ; Jiong WU ; Wenjia TANG ; Wenxin ZHU ; Baishen PAN
Chinese Journal of Laboratory Medicine 2009;32(10):1091-1095
Objective To investigate the urinary albumin excretion of the diabetes patients and application value in the monitoring of early impairment in kidney. Methods The random urine samples from diabetes patients and controls within three days were collectod. The changes of urinary albumin excretion within day and between days were analyzed. 24-hour urine albumin was used as a standard to evaluate early kidney damage. The correlations between results of random urine albumin at the different time points and different periods were comparod. The sensitivity and specificity of random urine albumin at the different time points and different periods was evaluated and compared to deduce the best diagnostic porformance of the random urine albumin. Results There are greater variations of the levels of urinary albumin of patients with diabetes and control. After the correction with urine creatinine and urine volume the variations can be reduced (CV:49%±23% and 64%±30%). Urinary albumin excretion rate change rhythmically within the 24 h in healthy and diabetes patients. We found the best correlation between overnight ratio of urinary concentrations of albumin and creatinine (ACR) and 24-hour urinary albumin (R2 = 0.976). It was superior to urina sanguinis (R~2 = 0.900), postprandial urine (R~2 = 0.584) and random urine (R~2 =0.791). When 24 h urinary albumin was taken as the standard, receiver operating characteristic (ROC) curve analysis showed there was significant difference between male and female(male 12.8 μg/mg urine creatinine vs female 27.0 μg/mg urine creatinine),and the the cut-off value of ACR was 27.7μg/mg urine creatinine. When the smallest available negative likelihood ratio (0.011) and the greatest positive likelihood ratio (481.000) were obtained,the concentration of 13.0 μg/mg creatinine and 87.4 μg/mg creatinine were set as the cut-off value of ACR. Conclusions The correction with urinary creatinine can reduce the variation between-days compared with urine volume, but still can not completely eliminate the variability. The ACR of overnight urine has the best correlation with the 24 h urinary albumin and can replace 24 h urinary albumin. Random urine as the most convenient collecting urine samples can also replace 24-hour urinary albumin, but the gender discrepancy need to be considered. When the concentration of 13.0 μg/mg and 87.4 μg/mg was set as a random ACR exclusion value and the confirmative value, it can basically rule out and confirm the existence of microalbuminuria.
8.Investigation on toxic shock syndrome toxin 1 from Staphylococcus aureus
Min WANG ; Xianping LI ; Jiong FU ; Jing FAN ; Aiguo TANG
Chinese Journal of Microbiology and Immunology 2008;28(4):334-337
Objective To detect the mecA gene and tst gene of toxic shock syndrome toxin 1 (TSST-1)of Staphylococcus aureus by using PCR and to learn the carrier condition of tst gene.Methods The mecA gene and tst gene of Staphylococcus aureus strains that isolated from clinical sources in our hospital during August 2006 to May 2007 were amplified in vitro using PCR,and to establish the rapid,specific,and sensitive method of detecting tst gene of methicillin resistant Staphylococcus aureus(MRSA).Results The mecA gene and tst gene were detected,and were made the gene sequencing successfully.Forty-one of 84 strains had mecA gene(48.81%),16 of 84 strains had tst gene(19.05%),10 of 84 strains had both of them,and the positive rate was 24.39%(10/41).Conclusion The proportion of tst gene positive strains of MRSA iS high in clinic,and it must be paid more attention.
9.Chinese herbal extracts inhibit the proliferation and induce the apoptosis of human lung cancer A549 cells
Qiuping LI ; Cong LIU ; Zuming TANG ; Qin CHEN ; Jiong LI
Journal of Medical Postgraduates 2016;29(3):240-244
Objective Cancer, a disease induced by abnormally regulated cell growth and apoptosis, is imposing a global threat to human health.This study was to explore the effects of Chinese herbal extracts ( CHE) in inducing the apoptosis and inhibiting the proliferation of human lung cancer cells. Methods Human lung cancer A549 cells were divided into a negative control, a high-dose CHE (680 ng/mL), a medium-dose CHE (340 ng/mL), and a low-dose CHE (170 ng/mL) group.The inhibitory effect of CHE on the proliferation of the lung cancer cells was detected by CCK8 and LDH assays, the apoptosis of the cells was assessed by fluorescence microscopy and flow cytometry, and the expressions of hTERT mRNA, cleaved caspase-3 and cleaved PARP were deter-mined by RT-PCR and Western blot. Results CHE inhibited the proliferation of the A549 cells with an IC50 value of 510 ng/mL. Treatment with high-dose CHE for 48 hours significantly suppressed the proliferation of the cells, induced the release of LDH, and promo-ted the apoptosis of the cells by 72.3%.RT-PCR and Western blot showed that 24-hour treatment with medium-dose CHE reduced the expression of hTERT mRNA by 4 times that of the negative control and up-regulated the expressions of cleaved caspase-3 and cleaved
PARP. Conclusion Chinese herbal extracts can induce cell apoptosis by decreasing the expression of hTERT mRNA and increasing those of the cleaved caspase 3 and cleaved PARP proteins.
10.The progress of clinical application of urinary albumin measurement
Wenjia TANG ; Jiong WU ; Wei GUO ; Baishen PAN
Chinese Journal of Laboratory Medicine 2014;(6):420-424
Urinary albumin used as an important indicator in diagnosis of renal disease , but there are still many problems in the clinical application .The urine specimen type , threshold value , reference measurement procedures and some more aspect were summarized in this article .