One-step detection of alpha fetal protein based on gold microelectrode through square wave voltammetry.

The detection of tumor markers in blood samples with high efficiency and sensitivity is in urgent need. In this work, a one-step quantitative detection assay for alpha fetal protein (AFP) based on gold microelectrode which is denoted as AuµE through square wave voltammetry using [Fe(CN)6]3-/4- as mediator was developed. As the biorecognition element of the assay, sulfydryl-modified AFP aptamer could be directly conjugated onto the surface of the AuµE, which could capture AFP with high specificity, and this attachment would cause the decrease of the capacitive current of the cyclic voltammetry due to the reduction of the active area of the electrodes. Under the optimized conditions, the AuµE aptasensor exhibited a linear detection range for AFP from 10-10 to 10-7 g/mL (S = 7.6 nA/dec, R2 = 0.991), and the detection limit is 2.5 × 10-11 g/mL. The AuµEs aptasensor demonstrates good selectivity against other types of proteins and small molecules, and has good reproducibility. The real blood samples were used for detection of AFP using the AuµEs aptasensor, the results agree well with those provided by the hospital through electrochemiluminescence method. Herein, the proposed one-step detection assay has a great application potential in point-of-care clinical diagnostics.

Effect of camrelizumab plus transarterial chemoembolization on massive hepatocellular carcinoma.

OBJECTIVE: To investigate the efficacy of camrelizumab plus transarterial chemoembolization (TACE) on massive hepatocellular carcinoma (HCC) patients. METHODS: A total of 92 cases with massive HCC from October 2019 to January 2021 were prospectively enrolled and randomly divided into the study group (n = 46) and the control group (n = 46). The control group received TACE while the study group were treated with camrelizumab plus TACE. The primary end points were clinical efficacy and adverse events. And the secondary end points were liver function, and alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9) levels before and after treatment. RESULTS: All participants were followed-up for 7 to 24 months, with a median of 12 months. Patients in the study group received TACE for 1-3 times, with an average of (2.01 ± 0.09) times, while patients in the control group receive TACE for 2-4 times, with an average of (3.78 ± 0.12) times, and the control group received significantly more TACEs (χ2 = 5.518, P = 0.019). During the follow-up, the response rate and disease control rate of the study group were significantly higher than those of the control group (χ2 = 5.518, P = 0.019; χ2 = 4.467, P = 0.041). Before treatment, the levels of total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alpha-fetoprotein (AFP), CEA, and CA19-9 were comparable between the groups (P > 0.05). After treatment, the levels of TBIL, ALT, AST, AFP, CEA, and CA19-9 decreased, and the above indicators in the study group were significantly lower than those in the control group (P < 0.05). All patients showed transient liver damage, vomiting, nausea, fever and abdominal pain after surgery, and their symptoms were relieved after symptomatic treatment. Adverse events occurred in 9 cases in the study group, and 3 cases in the control group (χ2 = 3.419, P = 0.064). CONCLUSION: Compared with TACE alone, camrelizumab plus TACE treatment can significantly improve the liver function of patients with massive HCC and enhance the treatment effect, which is worthy of clinical promotion.

Efficacy of Cyproheptadine Monotherapy in Hepatocellular Carcinoma With Bone Metastasis: A Case Report.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related deaths worldwide. Particularly, cases of bone metastasis have poorer prognoses. CASE PRESENTATION: A 62-year-old woman with suspected advanced HCC accompanied by bone metastasis with severe back pain and sciatica showed disease remission after cyproheptadine monotherapy. Initially, her serum alpha fetal protein (AFP) level was high, reaching up to 17697.62 ng/ml. A dose of 4 mg cyproheptadine, 3 times a day for 17 months was prescribed as the only treatment. Within 3 months, the serum AFP level gradually normalized down to 4.3 ng/ml. Both liver biopsy and bone biopsies were subsequently performed after 2 weeks of cyproheptadine. The results showed no malignancy. During the 34 months of follow-ups, the serum AFP remained normal in the range of 1.05 to 2.86 ng/ml. The patient has survived for 5 years without back pain and sciatica thus far. CONCLUSIONS: This is the first report to investigate a successful clinical approach in cyproheptadine monotherapy for an advanced HCC patient with bone metastasis. We recommend cyproheptadine as a potential anti-HCC agent for the treatment of HCC with bone metastasis, but more related studies such as prospectively clinical trials, and ideally randomized trials are still needed.

[The value of serum abnormal prothrombin in clinical application of hepatocellular carcinoma].

Objective: To examine the value of serum protein induced by vitamin K absence or antagonist-Ⅱ (PIVKA-Ⅱ) detection in the early diagnosis and surveillance of hepatocellular carcinoma (HCC). Methods: The clinical data of 215 patients with HCC admitted to Department of Hepatobiliary-Pancreatic Surgery of China-Japan Union Hospital of Jilin University from October 2017 to May 2018 were analyzed retrospectively. There were 172 males and 43 females, aged of (59.0±9.3) years old (range 34 to 86 years old). In addition, there were 85 non HCC patients were enrolled in the control group, 42 males and 43 females, aged (54.2±11.3) years old (range 22 to 80 years old). The blood sample of 3 ml was drawn from the elbow vein at 6∶00 am on the next day of admission, and then was kept in low temperature away from light, and sent for PIVKA-Ⅱ detection on the same day. The positive value of AFP was ≥20 µg/L and PIVKA-Ⅱ was ≥32 AU/L. The data were analyzed statistically by χ(2) test, t test or rank sum test. The correlation between AFP, PIVKA-Ⅱ and tumor maximum diameter was analyzed by linear regression. Results: The sensitivity of PIVKA-Ⅱ detection only for the diagnosis of HCC in all stages was significantly higher than AFP or equivalent to AFP, the overall sensitivity of PIVKA-Ⅱ and AFP was 85.1% and 52.1%, respectively. But the specificity of PIVKA-Ⅱ was lower than that of AFP, they were 78.8% and 96.5%, respectively. In particularly, in the earlier stage of HCC (Ⅰa) , the sensitivity of PIVAK-Ⅱ to HCC was 64.5%, while the AFP was only 26.3%. Combined detection of PIVKA-Ⅱ and AFP significantly improved the diagnostic rate of HCC to 88.4%, and the specificity to 76.5%. Moreover, there was a positive correlation between PIVKA-Ⅱ level and the maximum tumor diameter (r(2)=0.587, P<0.05), but there was no correlation between the AFP level and the maximum tumor diameter (r(2)=0.296, P>0.05). The positive rate of PIVKA-Ⅱ in the diagnosis of HCC with vascular invasion was also significantly higher than that of AFP (P<0.01) . Conclusions: PIVKA-Ⅱ can be used as a serological marker for HCC screening and diagnosis. In particular, PIVKA-Ⅱ detection was significantly sensitive than AFP in the earlier stage of HCC. Combined detection of PIVKA-Ⅱ and AFP can effectively improve the diagnostic rate of HCC in all stages. The significant elevation of PIVKA-Ⅱ is also helpful to determine the tumor aggressiveness, vascular invasion and prognosis of HCC patients.

Diagnosis and management of primary retroperitoneal teratoma in 36 infants / 中华实用儿科临床杂志

Objective To investigate the clinical manifestations and surgical effectiveness of the primary retroperitoneal teratoma in infants.Methods The records of 36 patients were retrospectively reviewed who were diagnosed as primary retroperitoneal teratoma and treated at the Department of Pediatric Oncology,Guangzhou Women and Children's Medical Center between August 2015 and August 2017.The related data were collected,including gender,age,operation time,operational bleeding and tumor weight.All patients took the ultrasound,computed tomography (CT) before surgery,alpha-fetal protein (AFP) and human chorionic gonadotrophin(HCG) were detected during peri-operation period.All cases were classified into 0 to Ⅲ grades according to the Norris classification (27 patients of 0 grade,1 patient of Ⅰ grade,4 patients of Ⅱ grade,4 patients of Ⅲ grade).Results There were 9 males and 27 females in the study.All patients were treated surgically at the age of 11.4 months (7 days-7 years) on the average;the mean weight of incised tumor was 736.47 g (7.90-2 355.00 g);operation time was 2.88 hours (1.08-7.06 hours);the amount of bleeding during operation was 29.11 mL (2-150 mL).All patients received follow-ups for 6.56 months (9 days-23 months) on the average.Radiographs found that tumors in 25 patients (69.4％) were located in the left side of the abdominal cavity,and the rest of them were located in the right side.In addition,tumors in 30 patients (83.3 ％) were big enough to pass across the mid-line of the body.Average pre-operation AFP was 7 593.1 μg/L(1.7-80 000.0 μg/L),post-operation AFP was 1 339.5 μg/L(1.4-16 519.6 μg/L),and the difference was statistically significant (P =0.001);pre-operation AFP of the mature group was 5 439.6 μg/L(1.7-80 000.0 μg/L),post-operation AFP of the mature group was 1 130.6 μg/L(1.4-16 519.6 μg/L),and the difference was statistically significant (P =0.001);pre-operation AFP of the immature group [11 182.3 μg/L(17.4-80 000.0 μg/L)] was higher than that of the mature group,and the difference was statistically significant(P =0.006).On the final follow-up,AFP was mean 38.3 μg/L (1.4-352.4 μg/L);among them,AFP of the mature group was mean 14.3 μg/L (1.4-43.3 μg/L),the immature group was 78.4 μg/L(1.7-325.4 μ g/L),and the difference was statistically significant(P =0.028);AFP of 4 patients in the immature group who underwent chemotherapy was 54.9 μg/L (2.6-116.6 μg/L)on the average,lower than those of post-operation AFP of 265.2 μg/L (206.8-384.1 μg/L),and the difference was statistically significant (P =0.042).All patients were treated surgically,and 3 cases of them received laparoscope technique;no significant relationship was proved between time of surgery and the tumor weight,location,and Norris classification(all P ＞ 0.05).The incidence rate of the surgical complications was 42.1％,such as tumor rupture (6 patients),vascular injury (5 patients),digestive tract damage (4 patients) and incomplete incision (1 patient).There was no death case in the serials.Conclusions Primary retroperitoneal teratoma can be completely incised.However,the complications of surgery could appear in many cases.As a predictive index for the recurrence retroperitoneal teratoma,AFP can be diminished by chemotherapy following the surgery.

Pt NPs and DNAzyme functionalized polymer nanospheres as triple signal amplification strategy for highly sensitive electrochemical immunosensor of tumour marker.

Highly sensitive determination of tumour markers is the key for early diagnosis of cancer. Herein, triple signal amplification strategy resulting from polymer nanospheres, Pt NPs, and DNAzyme was proposed in the developed electrochemical immunosensor. First, electroactive polymer nanospheres were synthesized by infinite coordination polymerization of ferrocenedicarboxylic acid, which could generate strong electrochemical signals due to plentiful ferrocene molecules. Further, the polymer nanospheres were functionalized by Pt NPs and DNAzyme (hemin/G-quadruplex) with the ability of catalyzing H2O2, which contributes to enhance the electrochemical signals. The prepared conjugations were characterized by transmission electron microscope (TEM) and energy dispersive X-ray spectroscopy (EDX). And the process of preparation was monitored by zeta potential. Based on the sandwich-type immunoassay, the electrochemical immunosensor was constructed employing the conjugations as signal tags. Under optimal conditions, the DPV peak increased with the increasing of alpha fetal protein (AFP) concentration, and the linear range was from 0.1pgmL(-1) to 100ngmL(-1) with low detection limit of 0.086pgmL(-1). Meanwhile, the designed immunosensor exhibited excellent selectivity and anti-interference property, good reproducibility and stability. More importantly, there were no significant differences in analyzing real clinical samples between designed immunosensor and commercial ELISA.

Effects of combined delivery of extremely low frequency electromagnetic field and magnetic Fe3O4 nanoparticles on hepatic cell lines.

Magnetic Fe3O4 nanoparticles (MNPs) have shown promise as drug carriers for treating lung and liver tumors in vivo. However, little is known about the combined delivery of these MNPs with a second approach, extremely low frequency electro-magnetic field (ELFF) exposure, which has been shown to have value for in vitro treatment of tumor cells. Here, ELFF and MNPs were combined to treat healthy (HL-7702) and cancerous (Bel-7402, HepG2) hepatic cells lines to explore the potential therapeutic effects, bio-mechanisms, and potential toxicity of a combined drug-free treatment in vitro. Flow cytometry for anti-AFP (alpha fetal protein) antibody, which coated the MNPs, indicated that the combined treatment induced Bel-7402 and HepG2 hepatoma cells lines into early apoptosis, without significant effects on healthy hepatic cells. This effect appeared to be mediated through cellular membrane ion metabolism. The presence of AFP-loaded MNPs strengthened the effects of ELFF on tumor cells, inducing a higher frequency of early apoptosis, while having minimal toxic effects on healthy HL-7702 cells. Western blotting revealed that the apoptosis-triggering BCL proteins were up regulated in hepatoma cells compared to healthy cells. Flow cytometry and patch-clamp studies revealed that this resulted from a higher MNP uptake ratio and greater cellular membrane ion exchange current in tumor cells compared to HL-7702 cells. Further, patch-clamp results showed that combining MNPs with ELFF treatment induces cells into early apoptosis through an ion metabolism disturbance in cells, similar to ELFF treatment. In brief, the combination of ELFF and MNPs had beneficial effects on tumor cells without significant toxicity on healthy cells, and these effects were associated with cellular MNP uptake.

Murine model of hepatic breast cancer.

BACKGROUND AND AIMS: Breast cancer is the most common cancer in women and the second leading cause of cancer-related deaths in this population. Breast cancer related deaths have declined due to screening and adjuvant therapies, yet a driving clinical need exists to better understand the cause of the deadliest aspect of breast cancer, metastatic disease. Breast cancer metastasizes to several distant organs, the liver being the third most common site. To date, very few murine models of hepatic breast cancer exist. METHODS: In this study, a novel murine model of liver breast cancer using the MDA-MB-231 cell line is introduced as an experimental (preclinical) model. RESULTS: Histological typing revealed consistent hepatic breast cancer tumor foci. Common features of the murine model were vascular invasion, lung metastasis and peritoneal seeding. CONCLUSIONS: The novel murine model of hepatic breast cancer established in this study provides a tool to be used to investigate mechanisms of hepatic metastasis and to test potential therapeutic interventions.

Hepatitis C patients serum HCV-RNA copy number and AFP-L3 percentage correlation analysis / 国际检验医学杂志

Objective To investigate the HCV patients with HCV‐RNA and AFP different plastid percentage (AFP‐L3/AFP) correlation ,analysis of liver cirrhosis to liver cancer caused by hepatitis c in the process of the development of HCV‐RNA and AFP‐L3/AFP change characteristics .Methods Collected clinical confirmed simple HCV patients 80 cases ,including 44 patients with cir‐rhosis of the liver and 36 cases of HCV liver cancer group and liver cirrhosis group were followed up at 3 ,6 months ,follow‐up re‐sults are divided into transfer of liver cancer group and treatment group ,in addition to choose 60 cases of healthy people as control group .Using case‐control study .Between different groups of HCV‐RNA and AFP‐L3/AFP compared by t test .Results The liver cirrhosis group average HCV‐RNA ,AFP‐L3/AFP for 7 .15 × 103 copy/mL ,44 .3% ,compared with the control group 400 copy/mL ,1 .1% ,the difference was statistically significant(P< 0 .05) .Liver cirrhosis group average HCV‐RNA ,AFP‐L3/AFP for 8 .33 × 109 copy/mL ,3 .35% ,and HCV liver cancer group (5 .71 × 107 copy/mL ,94 .33% ) ,the difference was statistically signifi‐cant(P<0 .05) .3 months follow‐up of patients with liver cirrhosis have 4 cases for the development of liver cancer ,8 cases after 6 months for the development of liver cancer .Conclusion The patients with cirrhosis caused by hepatitis c virus (HCV) should be combined ,dynamic detection of HCV RNA and AFP‐L3/AFP ,in order to early discover the liver cancer .

Role of plasma immunoglobulin receptor (PIgR/SC) in diagnosis of liver cancer / 中华内分泌外科杂志

Objective To investigate the diagnostic value of PIgP/SC in diagnosis of primary hepatic cancer.Methods 58 patients with primary hepatic cancer,60 patients with liver cirrhosis and 60 healthy volunteers were studied.4 ml fasting venous blood was collected from all subjects.Serum level of AFP was detected with electrochemical chemiluminescence immunoassay system and plasma PIgR/SC level was detected by ELISA method.The level of PIgR/SC and AFP was detected at one week after surgical resection in patients with hepatic cancer.Results The levels of AFP and PIgR/SC in the three groups were significantly different (P＜0.01),and PIgR/SC was higher than that in patients with cirrhosis and volunteers (P＜0.01).There was no significant difference between patients with liver cirrhosis and liver cirrhosis(P＞0.05).AFP was higher in patients with HCC than patients with cirrhosis and volunteers.AFP was higher in patients with cirrhosis than volunteers,and the difference was statistically significant(P＜0.01).Sensitivity of PIgR/SC and AFP was 89.3％ and 54.8％,specificity was 84.6％ and 91％,Youden index was 0.751 and 0.458,AUC was 0.920 and 0.761,respectively.There was significant difference in AUC (Z=3.251,P＜0.05) of the two detection indexes for detection of primary hepatic cancer.Conclusion The value of PIgR/SC in diagnosis of primary liver cancer may by higher than that of AFP.

Human alpha-fetal protein immunoassay using fluorescence suppression with fluorescent-bead/antibody conjugate and enzymatic reaction.

The aim of the study was to develop a simple and rapid immunoassay using fluorescent microbeads and enzyme-substrate reactions to measure alpha-fetal protein (AFP) concentrations. We demonstrated the functionality of the fluorescent immunosensor using antibody-conjugated fluorescent latex beads (AB-FLBs) and horseradish peroxidase (HRP) to catalyze a reaction, where the products would precipitate and suppress the fluorescence of AB-FLBs. First, the AB-FLBs were incubated with antigen, biotinylated antibodies (bABs), and streptavidin-HRP (SAv-HRP) to form a sandwich-type immunoreaction. The mixture was then filtered through a membrane to concentrate the beads on a small area. After washing to remove unbound bABs and SAv-HRP, a chromogenic HRP substrate and H2O2 were added to form precipitates on the FLB surface. The suppression of the fluorescence was measured with a fluorescent image analyzer system. Under optimized conditions, AFP could be measured at concentrations as low as 1 pg mL(-1) with a dynamic range up to 100 ng mL(-1).

Effect of glutamine on serum CEA, AFP and T cell immune function in postoperative patients with gastric cancer / 中国生化药物杂志

Objective To investigate the effect of glutamine on the serum carcino embryonie antigen (CEA) , alpha fetal protein (AFP) and T cell immune function in postoperative patients with gastric cancer.Methods 52 postoperative patients with gastric cancer from Yiwu Central Hospital were selected and randomly divided into control group and experimenal group, 26 cases in each group.The control group received routine nutritional therapy, and experimental group received alanyl -glutamine injection on the basis of control group.The serum nutritional parameters, CEA, AFP, CD4 +T, CD8 +T levels were compared post-treatment.Results Compared with control group post-treatment, the serum albumin (ALB) and pre-albumin (PA) levels were higher (P<0.05), the CEA and AFP levels were lower (P<0.05),the serum levels of CD4 +T and CD4 +T/CD8 +T were higher (P<0.05),CD8 +T level were lower (P<0.05).Conclusion Glutamine could significantly improve the clinical symptoms of patients with gastric cancer after surgery,and it is speculated that the mechanism may be the increase of serum ALB, PA levels, the decrease of CEA, AFP levels and the improvement of immune function.

Effect of magnesium sulfate combined with calcium channel blockers on cystatin C,uric acid, alpha fetal protein and urinary transferrin levels in patients with hypertension pregnancy / 中国生化药物杂志

Objective To explore the effect of magnesium sulfate combined with calcium channel blockers on cystatin C, uric acid, alpha fetal protein and urinary transferrin in patients with hypertension pregnancy.Methods 75 cases with hypertension of pregnancy were selected and divided into two groups, control group(n=37) were treated with conventional therapy, experiment group(n=38) were treated with magnesium sulfate combined with calcium channel blockers on the basis of control group.Cystatin C, uric acid, alpha fetal protein and urinary transferrin were compared after treatment between two groups.Results Compared with control group, Cys C, uric acid, serum AFP l, urinary TRF levels were all lower in experiment group( P<0.05), serum hematocrit, whole blood viscosity and erythrocyte aggregation index were all lower in experiment group(P<0.05).The total efficiency of experiment group was significantly higher than control group (P<0.05).Conclusion Magnesium sulfate combined with calcium channel antagonists in treatment of hypertensive disorder complicating pregnancy patients has good clinical curative effect, can effectively improve blood viscosity, reduce serum Cys C, UA, AFP and urinary TRF level.

The expression level of serum Golgi protein-73 in primary hepatocellular carcinoma and its clinical value in making early diagnosis / 介入放射学杂志

Objective To investigate the expression level of serum Golgi protein-73 (GP73) in patients with primary hepatocellular carcinoma (HCC), or accompanied with Budd-Chiari syndrome (BCS), and to discuss its clinical value in making early diagnosis. Methods The peripheral serum samples were collected in 490 individuals, including patients with HCC (n = 65), cholangiocarcinoma (CC, n = 35), Budd-Chiari syndrome accompanied with HCC (n = 40), type B virus hepatitis (n = 60), cirrhosis after B hepatitis (n = 60), Budd-Chiari syndrome complicated by cirrhosis (n = 60), other malignant lesions (n =50), benign hepatic lesions (n = 60) and healthy volunteers (n = 60). The expression levels of serum GP73 and AFP were determined by using Elisa and electrochemiluminescence methods separately. Results The expression level of serum GP73 in the patients with HCC , CC or BCS plus HCC were significantly higher than that in the other individuals (P 0.05). The area under ROC, sensitivity, specificity and the accuracy of GP73 for the diagnosis of CC were 0.774 , 88.6%, 72.7% and 73.9%respectively, and the optimal cut-off value was 45.40 ng/ml. Conclusion In diagnosing HCC, the expression level of serum GP73 is superior to AFP. It is hopeful that serum GP73 may be proved to be an early serum tumor marker for HCC.

The content change of AFP and AFP-L3 pre-and post-liver transplantation operation for liver cancer patients / 中国基层医药

Objective To explore the content change of Alpha Fetal Protein(AFP) and Alpha Fetal Protein Variants(AFP-L3) pre-and post-liver transplantation operation for liver cancer patients.Methods AFP-L3 was de-termined by micro centrifugal column from twelve cases of successive sera(For each case,one original shale pre-oper-ation,and mole shares post-operation successively),and preserved from patients who were diagnosed as fiver cancer and completed fiver transplantation operation.simultaneously the content of AFP and AFP-L3 in the original serum were also determined.and the ratio of AFP-L3 in AFP was calculated in order to compare the content change of AFP and the ratio of AFP-L3 pre-and post-liver transplantation operation.Results Compared with twelve cases of pre-and post-operation,the AFP in only five cases turned tO negative;The AFP in other cases keeped positive though AFP-L3%tumde to negative.and the content of AFP in some specific cases still keeped in a high level.Regarding AFP-L3,besides two casses turning to negative immediately after operation,the content of AFP-L3 in other patients de-creased gradually after operation,and it approached to negative(AFP-L3%<10%).Through statistical analysis,be-tween the half lives of AFP and AFP-L3%,there waa no statistic significance.Therefore,AFP-L3% could be considered as an independent reference item.Conclusion After a radical cure of liver resection or liver transplantation operation,AFP-L3%usually turned to negative in a rather short period (about 1 to 20 days).The ratio of AFP-L3 in AFP became be-low 10%,and it indicated that the operation would be quite complete,remaining liver would have hepatitis or hepatocirrho-sis.Otherwise,the ratio was not decrease during a long period,and it indicated the operation had not eliminated focuses completdy.