The role of Prolactin/Prolactin Receptor polymorphisms and expression in breast cancer susceptibility and outcome.

BACKGROUND: Prolactin (PRL) is a polypeptide hormone secreted by the anterior pituitary to stimulate growth and differentiation of the normal mammary gland. Together with its receptor, prolactin receptor (PRLR) have been shown to play a role in breast cancer. This study aimed to examine the roles of PRL and PRLR polymorphisms and expression in breast cancer risk and aggressiveness in Thai patients. METHODS: PRL (rs3756824 C/G and rs2244502 T/A) and PRLR (rs37364 G/T and rs249537 A/G) polymorphisms were genotyped by real-time PCR and PRLR expression was assessed by immunohistochemistry (IHC) in breast cancer tissues. The correlations between PRL and PRLR polymorphisms and breast cancer susceptibility/aggressiveness as well as the associations between PRLR expression and clinicopathological parameters were determined. RESULTS: Two hundred and twenty-seven breast cancer patients and 119 matched controls were recruited at the Division of Head Neck and Breast Surgery, Department of Surgery, Faculty of Medicine, Siriraj Hospital, Thailand from 2010-2014. PRL and PRLR polymorphisms were not correlated with breast cancer susceptibility and there was no association between PRLR polymorphisms and PRLR expression. PRLR was frequently overexpressed in breast cancer with positive hormone receptors. High expression of PRLR was significantly related to the presence of axillary nodal metastasis and lymphovascular invasion and showed a trend towards poorer outcome. CONCLUSIONS: There was a correlation between high PRLR expression and aggressive features of breast cancer. PRLR expression might be utilized as a prognostic factor for identification of luminal breast cancer with poorer outcome.

Study of the safety of oral Triphala aqueous extract on healthy volunteers.

BACKGROUND: Triphala extract is a well known medicinal herbal formula which is usually prescribed by Thai traditional doctors to adjust the physiological functions of the body. Previous studies have reported that Triphala has antioxidant, anti-inflammatory, antihypercholesterolemia and anticancer properties. Though this herbal recipe is commonly used in Thailand, its human safety, especially in the oral form, has not been studied. We therefore conducted a clinical trial (Phase I). OBJECTIVE: This study evaluated the safety of administering the aqueous extract of Triphala to healthy volunteers at 2500 mg/d. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: An open-label, single-arm trial was conducted at Chulabhorn International College of Medicine, Thammasat University, Pathum Thani, Thailand, between July 2017 and July 2018. The study enrolled 10 male and 10 female healthy volunteers; all were given Triphala (water extract; five capsules of 500 mg each) orally, once a day, at bedtime, for four consecutive weeks. MAIN OUTCOME MEASURES: Signs and symptoms, physical examinations, hematology and blood chemistry were assessed at the beginning of the trial and every week thereafter, for four consecutive weeks. After finishing the trial, on day 28, all volunteers were invited to a follow-up session on day 35 to evaluate the safety of the herbal recipe using the same measurements. RESULTS: At the oral dose of 2500 mg/d, Triphala had no serious adverse effects in healthy volunteers. Moreover, it was found to have significantly improved the volunteers' high-density lipoprotein cholesterol (HDL-C) levels on day 35 and also reduced their blood sugar levels on days 14 and 35. CONCLUSIONS: We conclude that aqueous extract of Triphala is safe for healthy volunteers and that it elevates HDL-C levels and lowers blood sugar. Further clinical study should investigate its effects on HDL-C and blood sugar levels among the dyslipidemic and prediabetic groups. TRIAL REGISTRATION: This trial was registered in the Thai Clinical Trial Registry with the identifier TCTR20180423002.

Pharmacokinetics of cucurbitacin B from Trichosanthes cucumerina L. in rats.

BACKGROUND: Cucurbitacin B is the major bioactive constituent in Trichosanthes cucumerina L. fruits, which the pharmacological properties have been studied for decades particularly an anti-tumor activity. The pharmacokinetic profile of this compound is still limited and investigation is needed for further phytopharmaceutical product development. This study aimed to investigate the pharmacokinetic profile of cucurbitacin B after administering the compound at different doses and routes to rats. METHODS: Male Wistar rats (n = 6) were treated by cucurbitacin B extracted from Trichosanthes cucumerina L. The cucurbitacin B was administered at 0.1 mg/kg intravenously or by oral gavage at 2-4 mg/kg. Blood samples and internal organs were collected serially within 24 h after administration. Urine and feces were collected from time 0 to 48 h. The level of cucurbitacin B in biological samples was determined by liquid chromatography-tandem mass spectrometry. RESULTS: The absolute oral bioavailability of cucurbitacin B was approximately 10%. The maximum concentration in plasma after normalization by dose ranged from 4.85-7.81 µg/L and the time to reach maximum value was approximately within 30 min after oral dosing. The level of cucurbitacin B in plasma increased proportionally to the given dose. After intravenous administration, cucurbitacin B had a large volume of distribution of about 51.65 L/kg and exhibited a high tissue to plasma concentration ratio, approximately 60 to 280-fold in several organs. Negligible amount of unchanged cucurbitacin B could be detected in urine and feces and accounted less than 1% of administered dose. CONCLUSION: Cucurbitacin B had low oral bioavailability, but could be distributed extensively into internal organs with a high volume of distribution and tissue to plasma ratio. Only negligible amounts of unchanged cucurbitacin B were excreted via urine and feces suggesting that the compound might be biotransformed before undergoing an excretion. Further studies of the metabolic pathway and tissue uptake mechanism are required to strategize the future development of cucurbitacin B into clinical studies.

Association of Estrogen Receptor Alpha and Interleukin 6 Polymorphisms with Lymphovascular Invasion, Extranodal Extension, and Lower Disease-Free Survival in Thai Breast Cancer Patients.

Breast cancer is the most frequent type of cancer diagnosed among women worldwide and also in Thailand. Estrogen and estrogen receptors exert important roles in its genesis and progression. Several cytokines have been reported to be involved in the microenvironment that promotes distant metastasis via modulation of immune and inflammatory responses to tumor cells. Estrogen receptor genetic polymorphisms and several cytokines have been reported to be associated with breast cancer susceptibility and aggressiveness. To investigate roles of genetic polymorphisms in estrogen receptor alpha (ESR1) and interleukin 6 (IL6), breast cancer patients and control subjects were recruited from the Division of Head, Neck and Breast Surgery (Siriraj Hospital, Bangkok, Thailand). Polymorphisms in ESR1 (rs3798577) and IL6 (rs1800795 and rs1800797) were evaluated by real-time PCR in 391 breast cancer patients and 79 healthy controls. Associations between genetic polymorphisms and clinicopathological data were determined. There was no association between genetic polymorphisms and breast cancer susceptibility. However the ESR1 rs3798577 CT genotype was associated with presence of lymphovascular invasion (OR=2.07, 95%CI 1.20-3.56, p=0.009) when compared to the TT genotype. IL6 rs1800795 CC genotype was associated with presence of extranodal extension (OR= 2.30, 95%CI 1.23-4.31, p=0.009) when compared to the GG genotype. Survival analysis showed that IL6 rs1800797 AG or AA genotypes were associated with lower disease-free survival. These findings indicate that polymorphisms in ESR1 and IL6 contribute to aggressiveness of breast cancer and may be used to identify high risk patients.

Antibacterial and Antiproliferative Activities of Plumericin, an Iridoid Isolated from Momordica charantia Vine.

Plumericin, an iridoid lactone, was isolated with relatively high yield from Momordica charantia vine using the supercritical fluid extraction (SFE) and the separation box (Sepbox) comprising dual combination of high-performance liquid chromatography and solid phase extraction. This compound showed antibacterial activity against Enterococcus faecalis and Bacillus subtilis with minimum inhibitory concentration (MIC) values better than cloxacillin. Plumericin potently inhibited proliferation of two leukemic cancer cell lines: they were acute and chronic leukemic cancer cell lines, NB4 and K562, with the effective doses (ED50) of 4.35 ± 0.21 and 5.58 ± 0.35 µg/mL, respectively. In addition, the mechanism of growth inhibition in both cell lines was induced by apoptosis, together with G2/M arrest in K562 cells.

Ribosomal protein L11- and retinol dehydrogenase 11-induced erythroid proliferation without erythropoietin in UT-7/Epo erythroleukemic cells.

Erythropoiesis is the process of proliferation, differentiation, and maturation of erythroid cells. Understanding these steps will help to elucidate the basis of specific diseases associated with abnormal production of red blood cells. In this study, we continued our efforts to identify genes involved in erythroid proliferation. Lentivirally transduced UT-7/Epo erythroleukemic cells expressing ribosomal protein L11 (RPL11) or retinol dehydrogenase 11 (RDH11) could proliferate in the absence of erythropoietin, and their cell-cycle profiles revealed G0/G1 prolongation and low percentages of apoptosis. RPL11-expressing cells proliferated more rapidly than the RDH11-expressing cells. The antiapoptotic proteins BCL-XL and BCL-2 were expressed in both cell lines. Unlike the parental UT-7/Epo cells, the expression of hemoglobins (Hbs) in the transduced cells had switched from adult to fetal type. Several signal transduction pathways, including STAT5, were highly activated in transduced cells; furthermore, expression of the downstream target genes of STAT5, such as CCND1, was upregulated in the transduced cells. Taken together, the data indicate that RPL11 and RDH11 accelerate erythroid cell proliferation by upregulating the STAT5 signaling pathway with phosphorylation of Lyn and cyclic AMP response element-binding protein (CREB).

Vascular endothelial growth factor ­634G/C polymorphism is associated with increased breast cancer risk and aggressiveness.

Polymorphisms in the promoter and 5' untranslated region of vascular endothelial growth factor (VEGF) have been associated with VEGF levels. To investigate the role of VEGF polymorphisms in breast cancer, the VEGF ­2578C/A, ­1498C/T, ­1154G/A and ­634G/C polymorphisms were genotyped in 483 breast cancer patients and 524 healthy controls. VEGF mRNA levels in breast cancer tissue were determined using semi­quantitative RT­PCR. The genotypes, ­634G/C and ­634C/C, were associated with an increased risk for breast cancer when compared with the ­634G/G genotype. The VEGF ­634G/C genotype was associated with tumor size >20 mm, perineural invasion and stage II­IV. Individuals with ­634C/C had lower disease­free survival. Patients with the VEGF ­634C/C genotype exhibited the highest VEGF mRNA levels. High VEGF mRNA expression correlated with tumor size >20 mm, presence of lymphovascular invasion and axillary nodal metastasis. These observations suggested that VEGF ­634G/C polymorphisms have a significant role in breast cancer susceptibility and aggressiveness.

Antiproliferative Effect and the Isolated Compounds of Pouzolzia indica.

Previous report showed the high potent antiproliferative effect of the methanolic part extracted from the aerial parts of Pouzolzia indica on NB4 and HT93A acute leukemic cell lines with the IC50 values of 28.5 and 49.8 µ g/mL, respectively. The bioassay-guided fractionation of the methanolic part gave 5 fractions, that is, FFI-FFV. FFII, FFIII, and FFIV inhibited the above leukemic cell lines with the IC50 values of 15.1 (FFII), 14.4 (FFIII), 32.1 (FFIV), and 31.0 (FFII), 9.7 (FFIII), 10.5 (FFIV) µ g/mL, respectively. The compounds in these fractions were isolated using chromatographic technique. FFII contained friedelin 1, 28-hydroxy-3-friedelanone 2, and 7-methoxy-coumarin 3. FFIII contained 6, 7-dimethoxy-coumarin 4, scopoletin 5, methyl caffeate 6. FFIV contained sitosteryl glucoside 7 and a supposed glycosphingolipid 8. The chemical structures were elucidated by spectroscopic methods.

Significant increase in cytotoxic T lymphocytes and natural killer cells by triphala: a clinical phase I study.

Background. Searching for drugs or herbal formulations to improve the immunity of HIV/AIDS positive people is an important issue for researchers in this field. Triphala, a Thai herbal formulation, is reported to have immunomodulatory effects in mice. However, it has not yet been investigated for immunostimulatory and side effects in healthy human volunteers. Objective. To evaluate the immunostimulatory and side effects of Triphala in a clinical phase I study. Materials and Methods. All volunteers took Triphala, 3 capsules per day for 2 weeks. Complete physical examination, routine laboratory analysis, and immunological studies were performed before ingestion and after initial meeting for 4 consecutive weeks. Results. We found that Triphala demonstrated significant immunostimulatory effects on cytotoxic T cells (CD3(-)CD8(+)) and natural killer cells (CD16(+)CD56(+)). Both of them increased significantly when compared with those of the control samples. However, no significant change in cytokine secretion was detected. All volunteers were healthy and showed no adverse effects throughout the duration of the study. Conclusion. Triphala has significant immunostimulatory effects on cellular immune response, especially cytotoxic T cells and natural killer cells. Increases in the absolute number of these cells may provide a novel adjuvant therapy for HIV/AIDS positive people in terms of immunological improvement.

Cutaneous involvement by colonic extranodal NK/T-cell lymphoma mimicking mycosis fungoides: a case report*.

We report a 51-year-old woman with cutaneous involvement by extranodal NK/T-cell lymphoma (TCL) of the colon that microscopically mimicked mycosis fungoides (MF). She had a history of fever of unknown origin for 2 months and then developed multiple erythematous papules on her trunk and extremities. A skin biopsy revealed superficial infiltration by atypical small to medium-sized lymphocytes with epidermotropism and Pautrier collections. Immunohistochemical studies showed expression of CD3 and TIA-1 with lack of expression (double negative) of CD4 and CD8. Initially, we reported the diagnosis as MF, cytotoxic variant. Thereafter, computerized tomography scan incidentally identified a colonic mass. A colonic biopsy revealed infiltration of atypical lymphoid cells with the same morphology and immunophenotype as those found in the skin. Additionally, CD56 and Epstein-Barr virus-encoded RNA in situ hybridization in both skin and colonic biopsies were diffusely positive. Thus, extranodal NK/TCL was diagnosed. Delta T-cell receptor (TCR) gene rearrangement was documented in the skin biopsy by polyacrylamide gel electrophoresis and fluorescence capillary gel electrophoresis methods. There was no TCR gene rearrangement detected in the colonic biopsy. Unfortunately, the patient died within 2 months of diagnosis.

Aberrant antigenic expression in extranodal NK/T-cell lymphoma: a multi-parameter study from Thailand.

BACKGROUND: Extranodal NK/T-cell lymphoma, nasal type (ENKTL) is not common worldwide, but it is the most common T- and NK-cell lymphomas in many Asian countries. Immunophenotypic profiles were studied based on limited series. The authors, therefore, studied on ENKTL according to characterize immunophenotypic profiles as well as the distribution of EBV subtype and LMP-1 gene deletion. METHODS: By using tissue microarray (TMA), immunohistochemical study and EBV encoded RNA (EBER) in situ hybridization were performed. T-cell receptor (TCR) gene rearrangement, EBV subtyping, and LMP-1 gene deletion were studied on the available cases. RESULTS: There were 22 cases eligible for TMA. ENKTL were positive for CD3 (91%), CD5 (9%), CD7 (32%), CD4 (14%), CD56 (82%), TIA-1 (100%), granzyme B (95%), perforin (86%), CD45 (83%), CD30 (75%), Oct2 (25%), and IRF4/MUM1 (33%). None of them was positive for ßF1, CD8, or CD57. TCR gene rearrangement was negative in all 18 tested cases. EBV was subtype A in all 15 tested cases, with 87% deleted LMP-1 gene. Cases lacking perforin expression demonstrated a significantly poorer survival outcome (p = 0.008). CONCLUSIONS: The present study demonstrated TIA-1 and EBER as the two most sensitive markers. There were a few CD3 and/or CD56 negative cases noted. Interestingly, losses of CD45 and/or CD7 were not uncommon while Oct2 and IRF4/MUM1 could be positive in a subset of cases. Based on the present study in conjunction with the literature review, determination of PCR-based TCR gene rearrangement analysis might not be a useful technique for making diagnosis of ENKTL.

Comparison of Pouzolzia indica methanolic extract and Virkon against cysts of Acanthamoeba spp.

The present study was conducted to investigate the morphological and structural changes of Acanthamoeba cysts after being treated with various concentrations of Pouzolzia indica methanolic extract fraction 3 (methanol eluted) and Virkon solution. Changes in the Acanthamoeba cysts were detected by light microscopy, scanning electron microscopy and transmission electron microscopy. The results show Acanthamoeba cysts were killed by Pouzolzia indica methanolic extract fraction 3 at a concentration of 1:8 and by Virkon solution at a concentration of 0.25%, with a minimal cysticidal concentration (MCC) by 24 hours. Both agents caused similar structural damage to Acanthamoeba cysts in the same sequence. Step by step structural alterations occurred within the cyst. First, the cyst shrank, collapsed and had clumping of cytoplasmic stuctures inside the cyst walls. Second, the cysts began to bulge, swell, have a decrease in wrinkles in the cyst walls and spill the cytoplasmic contents into the environment. Finally, the cyst walls broke into small pieces. This study may be beneficial to compare with future studies of pharmaceutical agents against Acanthamoeba keratitis.

Detection of monoclonal immunoglobulin heavy chain gene rearrangement (FR3) in Thai malignant lymphoma by High Resolution Melting curve analysis.

UNLABELLED: Malignant lymphoma, especially non-Hodgkin lymphoma, is one of the most common hematologic malignancies in Thailand. The diagnosis of malignant lymphoma is often problematic, especially in early stages of the disease. Detection of antigen receptor gene rearrangement including T cell receptor (TCR) and immunoglobulin heavy chain (IgH) by polymerase chain reaction followed by heteroduplex has currently become standard whereas fluorescent fragment analysis (GeneScan) has been used for confirmation test. In this study, three techniques had been compared: thermocycler polymerase chain reaction (PCR) followed by heteroduplex and polyacrylamide gel electrophoresis, GeneScan analysis, and real time PCR with High Resolution Melting curve analysis (HRM). The comparison was carried out with DNA extracted from paraffin embedded tissues diagnosed as B- cell non-Hodgkin lymphoma. Specific PCR primers sequences for IgH gene variable region 3, including fluorescence labeled IgH primers were used and results were compared with HRM. In conclusion, the detection IgH gene rearrangement by HRM in the LightCycler System showed potential for distinguishing monoclonality from polyclonality in B-cell non-Hodgkin lymphoma. INTRODUCTION: Malignant lymphoma, especially non-Hodgkin lymphoma, is one of the most common hematologic malignancies in Thailand. The incidence rate as reported by Ministry of Public Health is 3.1 per 100,000 population in female whereas the rate in male is 4.5 per 100,000 population 1. At Siriraj Hospital, the new cases diagnosed as malignant lymphoma were 214.6 cases/year 2. The diagnosis of malignant lymphoma is often problematic, especially in early stages of the disease. Therefore, detection of antigen receptor gene rearrangement including T cell receptor (TCR) and immunoglobulin heavy chain (IgH) by polymerase chain reaction (PCR) assay has recently become a standard laboratory test for discrimination of reactive from malignant clonal lymphoproliferation 34. Analyzing DNA extracted from formalin-fixed, paraffin-embedded tissues by multiplex PCR techniques is more rapid, accurate and highly sensitive. Measuring the size of the amplicon from PCR analysis could be used to diagnose malignant lymphoma with monoclonal pattern showing specific and distinct bands detected on acrylamide gel electrophoresis. However, this technique has some limitations and some patients might require a further confirmation test such as GeneScan or fragment analysis 56.GeneScan technique or fragment analysis reflects size and peak of DNA by using capillary gel electrophoresis. This technique is highly sensitive and can detect 0.5-1% of clonal lymphoid cells. It measures the amplicons by using various fluorescently labeled primers at forward or reverse sides and a specific size standard. Using a Genetic Analyzer machine and GeneMapper software (Applied Bioscience, USA), the monoclonal pattern revealed one single, sharp and high peak at the specific size corresponding to acrylamide gel pattern, whereas the polyclonal pattern showed multiple and small peak condensed at the same size standard. This technique is the most sensitive and accurate technique; however, it usually requires high technical experience and is also of high cost 7. Therefore, rapid and more cost effective technique are being sought.LightCycler PCR performs the diagnostic detection of amplicon via melting curve analysis within 2 hours with the use of a specific dye 89. This dye consists of two types: one known as SYBR-Green I which is non specific and the other named as High Resolution Melting analysis (HRM) which is highly sensitive, more accurate and stable. Several reports demonstrated that this new instrument combined with DNA intercalating dyes can be used to discriminate sequence changes in PCR amplicon without manual handling of PCR product 1011. Therefore, current investigations using melting curve analysis are being developed 1213.In this study, three different techniques were compared to evaluate the suitability of LightCycler PCR with HRM as the clonal diagnostic tool for IgH gene rearrangement in B-cell non-Hogdkin lymphoma, i.e. thermocycler PCR followed by heteroduplex analysis and PAGE, GeneScan analysis and LightCycler PCR with HRM.

Pouzolzia indica methanolic extract fraction 2 and povidone-iodine induced changes in the cyst of Acanthamoeba spp.: light and electron microscopic studies.

OBJECTIVE: To compare the minimal cysticidal concentration (MCC) between Pouzolzia indica methanolic extract fraction 2 and Povidone-lodine (PVP-I) on the Acanthamoeba cyst and to illustrate the morphological changes of the cyst after being treated by light and electron microscopies. MATERIAL AND METHOD: Acanthamoeba spp were isolated from patients with Acanthamoeba keratitis and cultured on a non-nutrient agar plate (NNA) seeded with heat killed Escherichia coli (NNA-E.coli) at 37 degrees C for 7 days, adjusted to a final concentration of 10(4) cysts/ml. Several concentrations of PVP-I and fraction 2 of Pouzolzia indica methanolic extract were tested to find the minimal cysticidal concentrations (MCC) of both agents, at these concentrations there was no viable cyst which was confirmed by no excystment after further incubation for 7 days. The cysts were prepared for routine transmission and scanning electron microscopic studies. RESULTS: Structural damages of the treated cysts by MCC of PVP-I and fraction 2 of Pouzolzia indica methanolic extract showed a series of damages. Starting from shrinkage, destruction or rupture of the cyst walls and opercula, withdrawal of the cytoplasm or edema cyst by the outside solution passed through the damaged wall caused a decrease in wrinkle ridges of the ectocyst. Then the cyst was ripped and torn into small pieces CONCLUSION: MCC of PVP-I solution and the fraction 2 of Pouzolzia indica methanolic extract were 0.04% and 1:4, respectively. The structural damages were somewhat similar, such as the shrinkage, ruptured cyst wall and opercula, edema and end by breaking up of the cyst wall and degeneration of the inside cytosol. Pouzolzia indica may be modified as an effective disinfectant solution for a contact lens case if the active ingredients are more purified.

Cytotoxic properties of root extract and fruit juice of Trichosanthes cucumerina.

The root extract of Trichosanthes cucumerina L. and bryonolic acid (1), its main constituent, as well as the fruit juice and cucurbitacin B (3), its main constituent, were tested for cytotoxicity against four human breast cancer cell lines (SKBR3, MCF7, T47D, and MDA-MB435), two lung cancer cell lines (A549 and SK-LU1), and one colon cancer cell line (Caco-2). The root extract had higher IC (50) values than bryonolic acid (1) against three breast cancer cell lines (MCF7 = 267/121, T47D = 316/124, MDA-MB435 = 140/90 microL/mL) and one lung cancer cell line (A549 = 106/100 microL/mL). The fruit juice also had higher IC (50) values than cucurbitacin B (3) against the four breast cancer cell lines (131/73, 375/35, 249/60, and 156/26 microL/mL, respectively) and one lung cancer cell line (141/41 microL/mL) as shown above, as well as against the colon cancer cell line (101/1.5 microL/mL). However, the root extract inhibited SK-LU1 more strongly than did the fruit juice, cucurbitacin B (3), and bryonolic acid (1) (149/169/180/>500 microL/mL, respectively). The root extract inhibited the two lung and three breast cancer cell lines (SKBR3, MDA-MB435, and MCF7) more strongly than the fruit juice. Bryonolic acid (1) inhibited MDA-MB435 somewhat better than the other tested human cancer cell lines. The fruit juice inhibited the colon cancer cell line (Caco-2) more strongly than the root extract. Cucurbitacin (3) inhibited human cancer cell lines, especially Caco-2, much more strongly than bryonolic acid (1). In addition to bryonolic acid (1), bryononic acid (2), cucurbitacin B (3), and dihydrocucurbitacin B (4) also were isolated from the root extract.

Prevalence of 60 kda and 52 kda Ro/SS-A autoantibodies in anti-Ro positive Thai sera in Siriraj Hospital.

BACKGROUND: Anti-Ro antibody may directly react against either Ro60 or Ro52 or both antigens. To be more applicable for routine laboratory practice, the specific antigen type for antibody detection should be identified before test application. OBJECTIVE: Investigate the prevalence of 60 kDa and 52 kDa Ro/SS-A antibodies in Thai patients' sera in Siriraj Hospital. MATERIAL AND METHOD: Specimens for anti-Ro were requested between June and December 2005. They were tested with EUROLINE test kit for prevalence determination. The principle of the test is a qualitative in-vitro-assay that contains test strips coated with parallel lines of 14 highly purified antigens. Of 84 specimens requested for anti-Ro antibody, 76 were collected and tested with the EUROLINE test kits and eight were excluded due to inadequacy. RESULTS: The prevalence of anti-Ro60 and anti-Ro52 of all sera tested for anti-Ro by EUROLINE test kit were 30% (95% CI: 20-40%) and 26% (95% CI: 16-36%), respectively; and, those in anti-Ro positive Thai sera were 82% (95% CI: 68-96%) and 71% (95% CI: 54-88%), respectively. The prevalence of anti-Ro52 alone in anti-Ro positive Thai sera and all specimens requested for anti-Ro was about 18% (95% CI: 4-32%) and 7% (95% CI: 1-13%), respectively. The agreement and Kappa value between the two methods were 0.9 and 0.77, respectively. The study suggests that the test for anti-Ro detection should provide both Ro 60 and Ro 52 antigens. CONCLUSION: The prevalence of both anti-Ro 60 and anti-Ro 52 were quite common, therefore, the test for this specific antibody should provide both antigens for antibody detection.

Antiproliferative effect of Erycibe elliptilimba on human breast cancer cell lines.

Erycibe elliptilimba Merr. & Chun., family Convolvulaceae, is a Thai traditional medicine which has long been prescribed for various infectious and malignant diseases. Bio-assays of extracts from Erycibe elliptilimba Merr. & Chun. showed that a fraction (fraction 3) from an methanolic extract had an antiproliferative effect on SKBR3 and MDA-MB435 human breast cancer cells. The ED50 value of Erycibe elliptilimba Merr. & Chun. fraction 3 was 56.07 and 30.61 microg/ml for SKBR3 and MDA-MB435, respectively. After 48 h of exposure, this fraction at a concentration of 100 microg/ml significantly reduced cell proliferation in both cancer cells. In MDA-MB435 cells, cell cycle analysis showed that the herb extract fraction 3 induced the accumulation of cells in G2/M phase, whereas no significant change in cell cycle was detected in SKBR3 cells. The results indicated that the extract fraction 3 could induce cell cycle arrest in some way. However, further investigation is needed to assess the molecular mechanisms mediated anticancer activities of this plant.

Antibacterial activities of four Thai medicinal plants.

Medicinal plants have long been used and prescribed in Thailand for centuries. Some of them have been used for treating various diseases including infectious diseases. Pouzolzia pentandra Benn., Gelonium multiflorum A. Juss., Erycibe elliptilimba Merr. and Chun., Balanophora abbreviate Bl. are Thai medicinal plants from the Thai pharmacopoeia that have been prescribed for treating unknown fevers including some specific infectious diseases. This investigation demonstrated the effects of these Thai medicinal plants for their antibacterial activities by using the macrodilution assay. Based on the present study, the water methanol fraction (fraction 2) of Balanophora abbreviate Bl. showed the antibacterial activity at the MIC level of 250 microg/ml but the activity was bacteriostatic in its effects. Therefore, the use of these medicinal plants in controlling fever and infectious diseases appears to be justified and further investigations may be required to obtain more information.

Molecular mechanism of herbs in human lung cancer cells.

Herbs have been considered natural and valuable sources for anticancer drug discovery. Herbal medicine has been prescribed in many countries over centuries for treating various diseases including infectious and malignant diseases. Nowadays, many of the drugs that have been used for treatment of malignant diseases are derived from natural products such as Taxol, a natural product isolated initially from Pacific Yew (Taxus brevifolia). This review article describes research on molecular mechanisms, especially cytotoxic effect of natural products from plant sources, primarily preclinical studies, involving human lung cancer cells in vitro for providing more knowledge and issues for potential drug development from medicinal herbs in the future.