Downregulation of Serum miR-133b and miR-206 Associate with Clinical Outcomes of Progression as Monitoring Biomarkers for Metastasis Colorectal Cancer Patients.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer in the world. Non-coding RNAs or microRNAs (miRNAs; miRs) biomarkers can play a role in cancer carcinogenesis and progression. Specific KRAS and EGFR mutation are associated with CRC development playing a role in controlling the cellular process as epigenetic events. Circulating serum miRs can serve for early diagnosis, monitoring, and prognosis of CRC as biomarkers but it is still unclear, clinically. OBJECTIVE: To determine potential biomarkers of circulating serum miR-133b and miR-206 in CRC patients Methods: Bioinformatic prediction of microRNA was screened followed by TargetScanHu-man7.2, miRTar2GO, miRDB, MiRanda, and DIANA-microT-CDS. Forty-four CRC serum (19 locally advanced, 23 distant advanced CRC) and 12 normal serum samples were subsequently extracted for RNA isolation, cDNA synthesis, and miR validation. The candidate circulating serum miR-133b and miR-206 were validated resulting in a relative expression via quantitative RT-PCR. Relative expression was normalized to the spike-internal control and compared to normal samples as 1 using the 2-ΔΔCt method in principle. RESULTS: Our results represented 9 miRs of miR-206, miR-155-5p, miR-143-3p, miR-193a-3p, miR-30a-5p, miR-30d-5p, miR-30e-5p, miR-543, miR-877-5p relate to KRAS-specific miRs, whereas, 9 miRs of miR-133b, miR-302a-3p, miR-302b-3p, miR-302d-3p, miR-302e, miR-520a-3p, miR-520b, miR-520c-3p and miR-7-5p relevance to EGFR-specific miRs by using the bioinformatic prediction tools. Our results showed a decreased expression level of circulating serum miR-133b as well as miR-206 associating with CRC patients (local and advanced metastasis) when compared to normal (P < 0.05), significantly. CONCLUSION: The circulating serum miR-133b and miR-206 can serve as significant biomarkers for monitoring the clinical outcome of progression with metastatic CRC patients. Increased drug-responsive CRC patients associated with crucial molecular intervention should be further explored, clinically.

Smart dual imprinted Origami 3D-ePAD for selective and simultaneous analysis of vanillylmandelic acid and 5-hydroxyindole-3-acetic acid carcinoid cancer biomarkers using graphene quantum dots coated with dual molecularly imprinted polymers.

Measuring the levels of the biomarkers vanillylmandelic acid (VMA) and 5-Hydroxyindole-3-acetic acid (5-HIAA) is a valuable tool for clinical diagnosis not only of neuroblastoma or carcinoid syndrome, but also of essential hypertension, depression, migraine, and Tourette's syndrome. Herein, we explore using graphene quantum dots (GQDs) coated with molecularly imprinted polymer (MIP) as novel dual-imprinted sensors for selective and simultaneous determination of VMA and 5-HIAA in urine and plasma samples. The dual-MIP was successfully coated on the GQDs core via co-polymerization of (3-aminopropyl) triethoxysilane (APTES) and tetraethyl orthosilicate (TEOS), acting as functional and cross-linking monomers, respectively. In addition, we successfully created the dual imprinted VMA and 5-HIAA shell on the GQDs' core via a one-pot synthesis. We fabricated a facile and ready-to-use Origami three-dimensional electrochemical paper-based analytical device (Origami 3D-ePAD) for simultaneous determination of VMA and 5-HIAA using a GQDs@dual-MIP modified graphene electrode (GQDs@dual-MIP/SPGE). The Origami 3D-ePAD was designed to form a voltammetric cell on a three-layer foldable sheet with several advantages. For example, they were quickly assembled and enhanced the device's physical durability with the hydrophobic backup sheet. The developed dual imprinted Origami 3D-ePAD leads to substantially enhanced sensitivity and selectivity to electrochemical signal amplification generated from increasing the electrode-specific surface area, electrocatalytic activity, and the large numbers of dual imprinted sites for VMA and 5-HIAA detection. The synthetic recognition sites are highly selective for 5-HIAA and VMA molecules with an imprinting factor of 8.46 and 7.10, respectively. Quantitative analysis relying on square wave voltammetry reveals excellent linear dynamic ranges of around 0.001-25 µM, with detection limits of 0.023 nM for 5-HIAA and 0.047 nM for VMA (3Sb, n = 3). The Origami 3D-ePAD provides high accuracy and precision (i.e., recovery values of 5-HIAA ranged from 82.98 to 98.40 %, and VMA ranged from 83.28 to 104.39 %), and RSD less than 4.37 %) in urine and plasma samples without any evidence of interference. Hence, it is well suited as a facile and ready-to-use disposable device for point-of-care testing. It is straightforward, cost-effective, reproducible, and stable. Furthermore, it allows for rapid analysis (analysis time ∼20s) useful in medical diagnosis and other relevant fields.

Facile and Compact Electrochemical Paper-Based Analytical Device for Point-of-Care Diagnostic of Dual Carcinogen Oxidative Stress Biomarkers through a Molecularly Imprinted Polymer Coated on Graphene Quantum-Dot Capped Gold.

Nanoscale imprinting significantly increases the specific surface area and recognition capabilities of a molecularly imprinted polymer by improving accessibility to analytes, binding kinetics, and template removal. Herein, we present a novel synthetic route for a dual molecularly imprinted polymer (dual-MIP) of the carcinogen oxidative stress biomarkers 3-nitrotyrosine (3-NT) and 4-nitroquinolin-N-oxide (4-NQO) as coatings on graphene quantum-dot capped gold nanoparticles (GQDs-AuNPs). The dual-MIP was successfully coated on the GQDs-AuNPs core via a (3-mercaptopropyl) trimethoxysilane (MPTMS) linkage and copolymerization with the 3-aminopropyltriethoxysilane (APTMS) functional monomer. In addition, we fabricated a facile and compact three-dimensional electrochemical paper-based analytical device (3D-ePAD) for the simultaneous determination of the dual biomarkers using a GQDs-AuNPs@dual-MIP-modified graphene electrode (GQDs-AuNPs@dual-MIP/SPGE). The developed dual-MIP device provides greatly enhanced electrochemical signal amplification due to the improved electrode-specific surface area, electrocatalytic activity, and the inclusion of large numbers of dual-imprinted sites for 3-NT and 4-NQO detection. Quantitative analysis used square wave voltammetry, with an oxidation current appearing at -0.10 V for 4-NQO and +0.78 V for 3-NT. The dual-MIP sensor revealed excellent linear dynamic ranges of 0.01 to 500 µM for 3-NT and 0.005 to 250 µM for 4-NQO, with detection limits in nanomolar levels for both biomarkers. Furthermore, the dual-MIP sensor for the simultaneous determination of 3-NT and 4-NQO provides high accuracy and precision, with no evidence of interference from urine, serum, or whole blood samples.

Candidate microRNAs as Biomarkers in Malaria Infection: A Systematic Review.

Malaria disease is a public health problem especially in tropical countries, 445.000 of malaria-related deaths have been reported in 2017. MicroRNAs (miRNAs) are small non-coding RNAs with 18-24 nucleotides in length, which have been demonstrated to regulate gene expression of several biological processes. The dysregulation of host immune-related gene expressions during the transcriptional process by microRNA has been extensively reported in malaria parasite invasion of erythrocytes infection. The candidate's miRNAs would be used as potential biomarkers in the future and perspective. A systematic review on miRNAs as candidate clinical biomarkers in malaria infection has been established in this study. Electronic databases (Medline, EMBASE, CINAHL and Cochrane data bases) were screened and articles were included as per established selection criteria. We comprehensively searched to identify publications related to malaria and miRNA. PRISMA guidelines were followed, 262 articles were searched, duplicates and unconnected papers were excluded. Nineteen articles were included in the study. It was found that malaria parasite infected liver or tissue produce tissue-specific miRNAs and release to the blood stream. The association of miRNAs including miR-16, miR-155, miR-150, miR-451 and miR-223 with the dysregulations of immune-related genes expression such as PfEMP-1, IFN-γ, AGO- 1 AGO-2; IL4, CD80, CD86, CD36, ANG-1 and ANG-2 during early, severe and/or cerebral malaria infections indicate the potential use of those miRNAs as biomarkers for malaria infection.

BURKHOLDERIA PSEUDOMALLEI BIOFILM PLAYS A KEY ROLE IN CHRONIC INFLAMMATION IN C57BL/6 MICE.

Burkholderia pseudomallei is a causative agent of melioidosis. Clinical signs of melioidosis vary from acute septicemia to chronic inflammation or subclinical infection. This study investigated the role of B. pseudomallei biofilm in chronic inflammation in lungs of infected C57BL/6 mice. Low doses of B. pseudomallei H777 and its biofilm defective M10 mutant were fed intra-gastrically to C57BL/6 mice and inflammatory responses were investigated by histopathological techniques. Two hundred colony forming units (CFUs) of B. pseudomallei H777 induced chronic inflammatory responses in mice on day 20 post-infection, with discrete interstitial infiltration by mononuclear inflammatory cells. On day 40 postinfection, there were marked thickening of alveolar septa and congested capillaries, which increased in severity by day 60. On the other hand, mice infected with B. pseudomallei M10 showed less mononuclear infiltration. The results indicate that B. pseudomallei defective in biofilm production gave rise to less severe pathology, resulting a higher rate of survival in infected mice; and pulmonary melioidosis could be developed in C57BL/6 mice by intra-gastric feeding makes it a possible animal model of chronic human melioidosis.

Relapsed Melioidosis Model in C57BL/6 Mice.

BACKGROUND: Burkholderia pseudomallei are the causative agents of melioidosis, a disease that has a high relapse rate in endemic areas. The mechanism of relapse is unclear OBJECTIVE: This study aimed to establish relapsed melioidosis in C57BL/6 mice by induction with B. pseudomallei. MATERIAL AND METHOD: Low doses of B. pseudomallei H777 and its biofilm defective mutant (M10) were intra-gastric fed to C57BL/6 mice. All the infected mice had suppressed immune status by intra-peritoneal injection of hydrocortisone at 2.5 mg per mouse at day 60 post-infection. Inflammatory response to the infection was investigated by histo-pathological studies and monitoring bacterial counts in the blood and organs. RESULTS: All the infected mice were found to have a high infiltration of mononuclear cells at day 60 post-infection. The results showed high bacterial counts in the blood in both strains post-suppressed immune status after two days. The biofilm mutant and wild type strains produced relapse in C57BL/6 mice but the latter was responsible for significantly more severe inflammation than the biofilm mutant. CONCLUSION: Low immune status may cause relapsed melioidosis in hosts with chronic inflammation.

Antibacterial Activity of Excretions-Secretions from Chrysomya megacephala Against Escherichia coli.

BACKGROUND: The blowfly, Chrysomya megacephala, is distributed worldwide. Previous studies found maggot excretions-secretions from other blowfly species inhibited pro-inflammatory response and antimicrobial activity. OBJECTIVE: This study aimed to test the bactericidal activity of excretions-secretions from C. megacephala larvae. MATERIAL AND METHOD: A total of 1,500 3-day-old larvae were used to collect excretions-secretions (ES) modified by the Barnes method. The bactericidal activity ofthe excretions-secretions was test by Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli using suitable liquid culture assay. Scanning electron microscope (SEM) was used to investigate the morphological change ofthe bacteria. RESULTS: E. coli were significantly inhibited by excretions-secretions from C. megacephala larvae. P. aeruginosa and S. aureus were not found to inhibit growth. CONCLUSION: The excretions-secretions from C. megacephala larvae may have a medical property for the inhibition of bacterial growth.

Prevalence of bla(PenA) and bla(OXA) in Burkholderia pseudomallei Isolated from Patients at Sappasitthiprasong Hospital and Their Susceptibility to Ceftazidime and Carbapenems.

BACKGROUND: Burkholderia pseudomallei is a causative agent of melioidosis. Ceftazidime is the preferred drug of choice for treatment. However, the motility rate is high in endemic areas. OBJECTIVE: This study aimed to determine the susceptibility tofour different antimicrobial agents and to detect the ß-lactamase genes in B. pseudomallei isolates from patients admitted to Sappasitthiprasong Hospital. MATERIAL AND METHOD: 85 B. pseudomallei isolates from patients admitted to Sappasitthiprasong Hospital between November 2010 and May 2011 were determined for antimicrobial susceptibility by standard disk diffusion and minimum inhibitory concentration (MIC). Real-time polymerase chain reaction (PCR) was used for the detection of bla(penA) and bla(OXA) in ß-lactamase genes. RESULTS: Almost all of the clinical isolates ofB. pseudomallei were susceptible to ceftazidime and imipenem. Cefatzidime MIC was ≤ 1-16 µg/ml and imipenem MIC was ≤ 1-4 µg/ml. The real-time PCR revealed that more than 90% of B. pseudomallei isolates carried bla(PenA) and bla(OXA). CONCLUSION: Although the clinical isolates of B. pseudomallei were susceptible to ceftazidime and imipenem, this study showed B. pseudomallei had a gene that produced beta-lactamase enzyme and may be poorly effective in the use of beta-lactam drugs.

Comparison of Automated and Conventional IHC Visual Scoring Analysis for MHC Class I and Tapasin Expression in Cervical Carcinoma.

BACKGROUND: Cervical cancer (CXCA) is the second most common cancer among women in Thailand and worldwide. Immune evasion caused by down-regulation of host immune responsive genes, such as MHC class I and loss of antigen processing machinery (APM), presents a capability leading to cancer development. Immunohistochemical staining (HC) is regarded as a common technique for protein marker detection in clinical laboratories. At present, IHC automation has been launched to facilitate the speed and feasibility to replace conventional IHC. However, evaluation of its use is still limited. OBJECTIVE: This study aimed to evaluate IHC scoring by automated visual analysis compared to conventional IHC analysis. MATERIAL AND METHOD: The paraffin-embedded tissues of 96 invasive CXCA were processed using a tissue microarray (TMA) platform followed by automated IHC staining of the anti-MHC class I (heavy chain, ß2M) and an APM-Tapasin expression. Conventional IHC and automated slide scanning with scoring visual analysis were compared. RESULTS: The results showed significant association between conventional and automated IHC evaluation (p-value > 0.05, Chi-square) for MHC class I and Tapasin stated in percentage of positive cancer cells, whereas intensity was found (p-value < 0.05, Chi-square) with moderate agreement (p-value < 0.001, kappa) 0.434-0.615 and 0.353-0.554, respectively. After calculated values, the results showed significant association between conventional and automated IHC evaluation (p-value > 0.05, Chi-square) for MHC class I and Tapasin with the highest agreement level (p-value < 0.001, kappa) of summation 0.595-0.755 and multiply scoring 0.633-0.689, respectively. CONCLUSION AND DISCUSSION: The automation softwarefor IHC scoring and interpretation can be used for the determination of MHC class I and Tapasin in CXCA. In addition, an antigen presentation pattern must be included to allow an accurate result for MHC class I in clinical use. An appropriate sample size and design of staging coverage as well as clinical prognosis outcomes of progression should be used infurther investigation.

The evaluation of loop-mediated isothermal amplification-quartz crystal microbalance (LAMP-QCM) biosensor as a real-time measurement of HPV16 DNA.

We have previously developed quartz crystal microbalance biosensor integrated with loop-mediated isothermal amplification (LAMP-QCM) for human papillomavirus (HPV) type58 DNA detection. Infection with HPV, particularly HPV16, remains a serious health problem due to its major risk factor contributing to cervical cancer. In the present study, LAMP-QCM biosensor was evaluated in terms of a quantitative assay for copy number of HPV16 DNA in cervical samples compared to quantitative PCR using TaqMan assay (TaqMan-qPCR). The detection limit of LAMP-QCM was found to be 10 fold more sensitive than TaqMan-qPCR with 100% specificity and 7.6% imprecision. Different plot of HPV16 DNA copy number using Bland-Altman analysis revealed 94% correlation between LAMP-QCM and qPCR. We therefore concluded that the developed LAMP-QCM biosensor provides a possible rapid and sensitive assay for HPV16 DNA quantification in a routine laboratory.

Antibiotic resistance patterns of Enterococcus spp. isolated from Musca domestica and Chrysomya megacephala in ubon Ratchathani.

BACKGROUND: The housefly Musca domestica and the blowfly Chrysomya megacephala are found worldwide and are medically significant as mechanical vectors of various pathogens from unsanitary locations to food, resulting in diseases in humans. OBJECTIVES: This study aimed to test the antimicrobial activity against Enterococcus spp. isolated from M. domestica and C. megacephala by standard disk diffusion and minimal inhibitory concentration (MIC), and to study the potential of M. domestica and C. megacephala to transfer multi-drug resistant enterococcus to humans. MATERIAL AND METHOD: Seven hundred adult flies were collected from fresh-food markets, garbage piles, restaurants, school cafeterias, and rice paddy fields in Muang Ubon Ratchathani and Warinchamrap in Ubon Ratchathani Province. Antimicrobial susceptibility for Enterococcus spp. isolated from adult flies was performed by disk diffusion test and minimum inhibitory concentration (MIC) determination. RESULTS: One hundred and twenty isolates of Enterococcus spp. were taken from 67 M. domestica and 53 C. megacephala. Standard disk diffusion showed the Enterococcus spp. isolates exhibited susceptibility to ampilcillin (99.2%), chloramphenicol (74.20%), tetracycline (75.0%), vancomycin (50.8%), and erythromycin (42.5%). The MICs of antimicrobial agents for all isolates were < or = 0.25-8 microg/mL for vancomycin, 1- > 16 microg/mL for tetracycline, 4- > 16 microg/mL for chloramphenicol, and 0.5-8 microg/mL for ciprofloxacin. CONCLUSION: The study demonstrated the potential of M. domestica and C. megacephala to carry Enterococcus spp. Nine antimicrobial susceptibility patterns were obtained among the 120 enterococci isolates.

Methicillin-resistant Staphylococcus aureus with reduced susceptibility to vancomycin in Sanprasitthiprasong Hospital.

BACKGROUND: Staphylococcus aureus is a species of bacteria that causes a number of diseases and more than 60% of it is presently resistant to methicillin. Vancomycin is the drug of choice for the eradication of methicillin-resistant S. aureus (MRSA). OBJECTIVE: This study aimed to investigate the susceptibility of heterogeneous vancomycin intermediate S. aureus (hVISA) and vancomycin intermediate S. aureus (VISA) to vancomycin by standard disk diffusion, microbroth dilution, a one-point population assay, and a population analysis profile. MATERIAL AND METHOD: Sixty-eight MRSA isolates from patients admitted to Sanprasitthiprasong Hospital between November 2010 and November 2011 were tested. RESULTS: Standard disk diffusion showed that all the MRSA isolates were susceptible to vancomycin. Vancomycin MICs for all isolates were 1-2 microg/mL. Only two MRSA isolates (2.9%) were able to grow on brain heart infusion agar supplemented with vancomycin 4 microg/mL and were confirmed by a population analysis as hVISA. CONCLUSION: This study showed the effect of vancomycin on MRSA and the need for early detection and controlled planning.

Prognostic factors of human papillomavirus genotypes of invasive cervical carcinoma: an analytical cross-sectional study in lower north-east Thailand.

BACKGROUND: Cervical cancer (CXCA) caused by persistent infections by high-risk human papillomavirus (HR-HPV) can lead to multi-step carcinogenesis. The best management strategy and significant prognosis for cervical cancer patients remain unclear. OBJECTIVE: To investigate the associations of the two most common HR-HPVs with clinical outcomes of progression and recurrence status as well as prognosis outcomes of patients. MATERIAL AND METHOD: An analytical cross-sectional study of patients registered at Ubon Ratchathani Cancer Hospital was conducted from 2007 to 2010. Clinical data, histopathological features, and clinical outcomes of progression and recurrence status were recorded. HPV type-specific E6/E7 nested multiplex polymerase chain reaction (NMPCR) was performed to identify HR-HPV16 and 18 using extracted deoxyribonucleic acid (DNA) from embedded paraffin. Clinical findings and HPV genotypes were analyzed using Fisher's exact test. Association studies of crucial factors and HR-HPV genotypes were performed using logistic regression analysis (odds ratio [OR]) and 95% confidence interval [CI]). A p-value of less than 0.05 was considered statistically significant. RESULTS: The study found single HPV16 infection in 57.3%, single HPV18 in 17.3%, mixed HR-HPV16/18 in 13.1%, and non-HPV16, 18, or 16/18 in 12.3%. The findings showed significant association among their genotypes and histopathological types and grading (p < 0.0001 and p = 0.014). Clinical outcomes of progression and recurrence status with increased severity of clinical staging were associated significantly (p = 0.001 and p = 0.002). HPV18 type-specific was shown as a poor prognostic type with its relevance to the severity of disease higher than that of HPV16. CONCLUSION AND DISCUSSION: HPV16 and 18 remain the major type-specifics especially in relation to invasive CXCA, requiring further therapeutic vaccination study and proper prognosis. HR-HPV type-specific is very important during cervical carcinogenesis but other crucial contributing factors for prognostic outcomes should be further elucidated.

The development of DNA-based quartz crystal microbalance integrated with isothermal DNA amplification system for human papillomavirus type 58 detection.

To address the effect of dramatic change in temperature and viscosity during PCR process on quartz crystal microbalance (QCM) sensor and to increase the sensitivity, isothermal amplification was employed in the system. We combined loop-mediated isothermal amplification (LAMP) technique with QCM, called as LAMP-QCM, for detection of high-risk human papillomavirus viral DNA type 58 (HPV-58) which is commonly found in Asian women. The liquid-phase LAMP-QCM prototype comprised the frequency counter, a temperature control device and housing of the quartz crystal with polished gold electrodes on both sides. QCM detection signal was monitored in real-time based on an avidin-biotin binding between avidin coated QCM surface and specific biotinylated LAMP products. Analytical performance was evaluated for precision, sensitivity and specificity. A plasmid clone containing the HPV-58 sequence was diluted from 10(6) to 1 copy and used for detection limit. Cut-off value was estimated at 28.8 Hz from negative viral template. The system could detect 100 copies with Δf at 34.0±3.6 Hz compared to 1000 copies detected by conventional LAMP. No cross-reaction was observed with other HPV types. The HPV-58 detection was compared among LAMP-QCM, conventional LAMP and nested PCR in 50 cervical cancer tissues. The positive rate of LAMP-QCM was higher than that of conventional LAMP with 100% sensitivity and 90.5% specificity. The integrated LAMP-QCM system has improved the detection limit up to ten times compared to conventional LAMP with less-time consuming.

Activated charcoal enhanced the antigen-expression and dendritic cell maturation of the vaccine using Listeria-platform.

BACKGROUND: Listeria monocytogenes (LM) has been used as a vaccine vector based upon its ability to induce a strong cell-mediated immune response. LM inactivated with γ-irradiation retains immunogenic properties and is an attractive platform for clinical use since it would have improved safety concerns compared to live vectors. Activated charcoal has been shown to enhance expression of LM proteins such as PrfA. AIM: To investigate the effect of various growth conditions supplemented with activated charcoal on recombinant antigen expression. METHODS: We prepared γ-irradiated ovalbumin-expressing LM (LM-OVA) after growth under various culture conditions. We cultured LM-OVA at various temperatures including 25°C, 37°C and 37°C with activated charcoal and compared OVA expression by western blot analysis, dendritic cells maturation and OVA-specific T cells. RESULTS: The OVA expression was highest in γ-irradiated LM-OVA grown with activated charcoal at 37°C. Compared to other growth conditions, γ-irradiated LM-OVA grown with activated charcoal at 37°C induce better DC maturation as well as production of the highest number of antigen-specific IFN γ-secreting T cells. CONCLUSION: The further study should be demonstrated the potential to alter growth conditions to enhance OVA expression resulting for vaccine vectors, thereby improving their safety and efficacy.

The use of viral load as a surrogate marker in predicting disease progression for patients with early invasive cervical cancer with integrated human papillomavirus type 16.

OBJECTIVE: The purpose of this study was to assess the effectiveness of the use of human papillomavirus type 16 (HPV16) physical status and viral load in combination to predict clinical outcome during cervical development. STUDY DESIGN: A follow-up study was monitored in association with HPV integration and viral load in 121 cervical samples with the use of multiplex quantitative polymerase chain reaction. RESULTS: A significant increase of viral load was found earlier from preinvasive to invasive groups compared with normal groups, except with clinical staging and clinical outcome. High occurrence of integrated HPV16 was observed in preinvasive (27/44 samples) and invasive cervical carcinoma (40/68 samples). Cervical progression was observed significantly in most preinvasive (18/27 samples) and invasive cases (25/40 samples) that were infected with integrated HPV. Integrated HPV16 with significant viral load can be used as a predictive marker for tumor progression in the early stage of invasive cervical carcinoma. CONCLUSION: Integrated HPV16 in combination with viral load is a predictive indicator for tumor progression in early invasive stage but not in preinvasive and advanced invasive stage.