Anti-proliferative and apoptotic effect of cannabinoids on human pancreatic ductal adenocarcinoma xenograft in BALB/c nude mice model.

Human pancreatic ductal adenocarcinoma (PDAC) is a highly malignant and lethal tumor of the exocrine pancreas. Cannabinoids extracted from the hemp plant Cannabis sativa have been suggested as a potential therapeutic agent in several human tumors. However, the anti-tumor effect of cannabinoids on human PDAC is not entirely clarified. In this study, the anti-proliferative and apoptotic effect of cannabinoid solution (THC:CBD at 1:6) at a dose of 1, 5, and 10 mg/kg body weight compared to the negative control (sesame oil) and positive control (5-fluorouracil) was investigated in human PDAC xenograft nude mice model. The findings showed that cannabinoids significantly decreased the mitotic cells and mitotic/apoptotic ratio, meanwhile dramatically increased the apoptotic cells. Parallelly, cannabinoids significantly downregulated Ki-67 and PCNA expression levels. Interestingly, cannabinoids upregulated BAX, BAX/BCL-2 ratio, and Caspase-3, meanwhile, downregulated BCL-2 expression level and could not change Caspase-8 expression level. These findings suggest that cannabinoid solution (THC:CBD at 1:6) could inhibit proliferation and induce apoptosis in human PDAC xenograft models. Cannabinoids, including THC:CBD, should be further studied for use as the potent PDCA therapeutic agent in humans.

Effectiveness of Favipiravir monotherapy in the treatment of COVID-19: real world data analysis from Thailand.

Background: Previous studies showed that Favipiravir, a selective viral ribonucleic acid dependent-ribonucleic acid polymerase inhibitor, exhibited a trend of clinical improvement within 14 days and promoted viral clearance by day 7, without reduction of mortality rate in COVID-19. Methods: During the COVID-19 pandemic, Department of Medical Services (Thailand) formulated National Clinical Treatment Guidelines for COVID-19 and approved Favipiravir to eight medical centres. After treatment with Favipiravir monotherapy, we compared real-world data analysis to supportive treatment without antiviral agents. Findings: We analysed 12,888 COVID-19 patients between June 1, 2021, and July 31, 2021. This group study excluded 66 asymptomatic and 4634 COVID-19 patients treated with other antiviral agents. The 4896 mild, 2357 moderate, and 935 severe COVID-19 patients were analysed. All patients neither had previous SARS-CoV-2 infection nor received an mRNA vaccine during study period. Favipiravir monotherapy reduced the 28-day mortality risk in severe COVID-19 by relative risk (RR) = 0.72 (95% CI 0.58-0.91 P = 0.006) after adjustment for aging and hypertension. However, in mild and moderate COVID-19, Favipiravir monotherapy did not significantly reduce 28-day mortality risk by RR = 0.59 (95% CI 0.06-5.43 P = 0.65) after adjustment for aging, and RR = 0.60 (95% CI 0.32-1.13 P = 0.11) after adjustment for aging and obesity, respectively. In the patient with recovery, Favipiravir monotherapy exhibited a shortening time to recovery when compared to supportive treatment without antiviral agents (mean ± SD by 9.6 ± 7.1 vs. 12.9 ± 7.6 days: P < 0.0001, 10.0 ± 5.9 vs. 12.4 ± 5.3 days: P < 0.0001, and 11.2 ± 7.8 vs. 13.1 ± 8.0 days: P < 0.0001 in mild, moderate, and severe COVID-19 respectively). Interpretation: Real-world data analysis showed that favipiravir monotherapy was superior to supportive treatment without antiviral agents in shortening the recovery time in surviving patients and significantly reducing 28-day mortality risk in severe COVID-19. Funding: Department of Medical Services, Ministry of Public Health, Thailand.

Molecular characterisation and tracking of severe acute respiratory syndrome coronavirus 2 in Thailand, 2020-2022.

The global COVID-19 pandemic, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was first detected in China in December 2019. To date, there have been approximately 3.4 million reported cases of COVID-19 and over 24,000 deaths in Thailand. In this study, we investigated the molecular characteristics and evolution of SARS-CoV-2 in Thailand from 2020 to 2022. Two hundred sixty-eight SARS-CoV-2 isolates, collected mostly in Bangkok from COVID-19 patients, were characterised by partial genome sequencing. Moreover, the viruses in 5,627 positive SARS-CoV-2 samples were identified as viral variants - B.1.1.7 (Alpha), B.1.617.2 (Delta), B.1.1.529 (Omicron/BA.1), or B.1.1.529 (Omicron/BA.2) - by multiplex real-time reverse transcription polymerase chain reaction (RT-PCR) assays. The results revealed that B.1.36.16 caused the predominant outbreak in the second wave (December 2020-January 2021), B.1.1.7 (Alpha) in the third wave (April-June 2021), B.1.617.2 (Delta) in the fourth wave (July-December 2021), and B.1.1.529 (Omicron) in the fifth wave (January-March 2022). The evolutionary rate of the viral genome was 2.60 × 10-3 (95% highest posterior density [HPD], 1.72 × 10-3 to 3.62 × 10-3) nucleotide substitutions per site per year. Continued molecular surveillance of SARS-CoV-2 is crucial for monitoring emerging variants with the potential to cause new COVID-19 outbreaks.

Quantitative STAU2 measurement in lymphocytes for breast cancer risk assessment.

Although mammograms play a key role in early breast cancer detection, the test is not applicable to all women, for example, women under the age of 40. The development of a noninvasive blood test with high sensitivity and accessibility will improve the effectiveness of breast cancer screening programmes. Secretory factors released from cancer cells can induce the expression of certain genes in a large number of white blood cells (WBCs). Therefore, cancer-dependent proteins in WBCs can be used as tumour markers with high sensitivity. Five proteins (LMAN1, AZI2, STAU2, MMP9 and PLOD1) from a systemic analysis of a variety of array data of breast cancer patients were subjected to immunofluorescence staining to evaluate the presence of fixed WBCs on 96-well plates from 363 healthy females and 358 female breast cancer patients. The results revealed that the average fluorescence intensity of anti-STAU2 and the percentage of STAU2-positive T and B lymphocytes in breast cancer patients (110.50 ± 23.38 and 61.87 ± 12.44, respectively) were significantly increased compared with those in healthy females (56.47 ± 32.03 and 33.02 ± 18.10, respectively) (p = 3.56 × 10-71, odds ratio = 24.59, 95% CI = 16.64-36.34). The effect of secreted molecules from breast cancer cells was proven by the increase in STAU2 intensity in PBMCs cocultured with MCF-7 and T47D cells at 48 h (p = 0.0289). The test demonstrated 98.32%, 82.96%, and 48.32% sensitivity and 56.47%, 83.47%, and 98.62% specificity in correlation with the percentage of STAU2-positive cells at 40, 53.34 and 63.38, respectively. We also demonstrated how to use the STAU2 test for the assessment of risk in women under the age of 40. STAU2 is a novel breast cancer marker that can be assessed by quantitative immunofluorescence staining of fixed WBCs that are transportable at room temperature via mail, representing a useful risk assessment tool for women without access to mammograms.

Frequency and Association Of GSTM1 and GSTT1 Gene Polymorphisms with Survival in Breast Cancer Patients.

OBJECTIVE: Glutathione S-transferase M1 and T1 (GSTM1 and GSTT1) are the key detoxification enzymes of xenobiotics, including chemotherapeutic drugs. The deletion polymorphisms of GSTM1 and GSTT1 genes are associated with reduced enzyme activity that influenced clinical outcomes of chemotherapeutic agents in breast cancer. However, there is limited information among Thai patients. This research aims to explore the frequency and role of GSTM1 and GSTT1 polymorphisms on survival among Thai patients with breast cancer. METHODS: The retrospective cohort study was performed. Demographic data and clinicopathology characteristics were collected from hospital base registry data and medical records. A multiplex qualitative real-time PCR method was used to detect the presence or absence of the GSTM1 and GSTT1 gene in the genomic DNA samples of the participants. RESULTS: The frequencies of the GSTM1 and GSTT1 null genotypes in 198 breast cancer patients were 65.70% and 33.30%, respectively. The overall survival at 1, 3 and 5 years were 95.00%, 83.00%, 71.00% respectively. The log rank test and Cox proportional hazards revealed a significant different in the 5-years overall survival according to lymph node metastasis and tumor stage (P = 0.014 and P < 0.001). No associations between overall survival and GSTM1 or GSTT1 genotype were found in single or combined genotypes analyses (P = 0.76 and P= 0.15). CONCLUSION: The results of our study provided the epidemiological information for prognostic of survival in breast cancer patients treated with chemotherapy.
.

Negative Impact of 25-hydroxyvitamin D Deficiency on Breast Cancer Survival.

BACKGROUND: Low 25-hydroxyvitamin D [25(OH)D] levels in serum are associated with breast cancer risk. This study was conducted to determine the impact of 25(OH)D deficiency on survival of breast cancer patients. METHODS: In a retrospective cohort study of 303 patients diagnosed with breast cancer during 2011-2012 at the National Cancer Institute Thailand, all cases were followed up for 7 years. The 25(OH)D was measured by high-performance liquid chromatography (HPLC). Clinical and pathological data were collected. The Chi-square test, Kaplan-Meier and Cox regression model were used to assess the association between 25(OH)D levels and risk of death. RESULTS: Of the 303 cases aged between 24 and 78 years 51 (16.8%) died during follow-up from any cause. The mean 25(OH)D levels was 25.1±7.54 ng/ml (8.2 - 61.0 ng/ml). Thirty-three patients (10.9%) were stratified as inadequate or deficient group (<16 ng/ml) with mean survival time of 60.65 months compared to 76.24 months in insufficient or sufficient group (≥16 ng/ ml). Multivariate analysis adjusted for age, body mass index, stage, lymph node metastases, and immunohistochemical (IHC) findings (ER, PgR, HER-2, Ki-67 and P53) showed that patients with low 25(OH)D levels (<16 ng/ml) at diagnosis had a significantly higher risk of death (hazard ratio = 2.5-2.9) than the group with high 25(OH)D levels (≥16 ng/ ml). CONCLUSION: A concentration of 25(OH)D below 16 ng/ml was found to be independently associated with poor survival in breast cancer patients, regardless of age, lymph node status, stage or breast cancer subtype. An investigation of potential benefit of 25(OH)D supplements appears warranted.

Impact of Using Intra-Operative Ultrasound Guided Breast- Conserving Surgery on Positive Margin and Re-Excision Rates in Breast Cancer Cases with Current SSO/ASTRO Guidelines.

PURPOSE: To review the impact of using intra-operative ultrasound guided breast conserving surgery with frozen sections on nal pathological margin outcome with the current guidelines set forth by the Society of Surgical Oncology (SSO) and the American Society of Surgical Oncology (ASTRO). MATERIALS AND METHODS: A retrospective review including all cases of intra-operative ultrasound guided breast conserving surgery was performed at the National Cancer Institute Thailand between 2013 and 2016. Patient demographics, tumor variables, intraoperative frozen section and nal pathological margin outcomes were collected. Factors for positive or close margins were analyzed. RESULTS: A total of 86 patients aged between 27 and 75 years with intra- operative ultrasound guided breast conserving surgery were included. Three cases (3.5%) of positive margin were detected by intra-operative frozen section and 4 cases (4.7%) by final pathology reports. There were 18 cases (20.9%) with a close margin (<1 mm). Factors affecting this result comprised multi-foci, presence of invasive ductal carcinoma (IDC) combined with ductal carcinoma in situ (DCIS) and invasive lobular carcinoma (ILC). CONCLUSIONS: With the current SSO/ASTRO for adequate margin guidelines, using intra-operative ultrasound to locate the boundary for resection with breast conserving surgery provided a high success rate in obtaining final pathology free margin outcomes and minimizing re-operation risks especially when combined with intra-operative frozen section assessment. The chance of finding positive or close margins appears higher in cases of IDC combined with DCIS, ILC and with multi-foci cancers.

Association of Vitamin D Level with Clinicopathological Features in Breast Cancer.

A population-based relationship between low vitamin D status and increased cancer risk is now generally accepted. However there were only few studies reported on prognostic impact. To determine the effect of low vitamin D on progression of breast cancer, we conducted a cross-sectional analysis of vitamin D levels and clinico- pathological characteristics in 200 cases of breast cancer diagnosed during 2011-2012 at the National Cancer Institute of Thailand. Vitamin D levels were measured by high-performance liquid chromatography (HPLC). Clinical and pathological data were accessed to examine prognostic effects of vitamin D. We found that the mean vitamin D level was 23.0±6.61 ng/ml. High vitamin D levels (≥32 ng/ml) were detected in 7% of patients, . low levels (<32 ng/ml) in 93% Mean vitamin D levels for stages 1-4 were 26.1±6.35, 22.3±6.34, 22.2±6.46 and 21.3±5.42 ng/ml respectively (P=0.016) and 24.1 and 21.3 ng/ml for lymph node negative and positive cases (P=0.006). Low vitamin D level (<32 ng/ml) was significantly found in majority of cases with advanced stage of the disease (P=0.036), positive node involvement (P=0.030) and large tumors (P=0.038). Our findings suggest that low and decreased level of vitamin D might correlate with progression and metastasis of breast cancer.

Clinical outcome of breast cancer BI-RADS 4 lesions during 2003-2008 in the National Cancer Institute Thailand.

To determine the clinical outcome of breast cancer BI-RADS 4 lesions and seek a more effective management guideline, we conducted a retrospective study of all BI-RADS4 patients diagnosed between 2003-2008 with follow up time not less than 2 years. A total of 392 cases of BI-RADS 4 were identified and 320 could be sub-categorised as 4a, 4b and 4c. Overall malignant positive results were 7.65, 38.7 and 58.percent, respectively. In all cases assigned to the close follow up group, no malignancy was detectable (P<0.02). The results of the study suggested that BI-RADS sub-categories have benefit for cancer diagnosis and treatment decisions of clinicians and it might be possible to set up a safe follow-up guideline in selected groups of patients to minimize un-necessary tissue biopsy for breast cancer detection.