Differentiation capacity of dental pulp stem cell into inner ear hair cell using an in vitro assay: a preliminary step toward treating sensorineural hearing loss.

PURPOSE: Sensorineural hearing loss (SNHL) is commonly caused by the death or dysfunction of cochlear cell types as a result of their lack of regenerative capacity. However, regenerative medicine, such as stem cell therapy, has become a promising tool to cure many diseases, including hearing loss. In this study, we determined whether DPSCs could differentiate into cochlear hair cell in vitro. METHODS: DPSCs derived from human third molar dental pulp were induced into NSCs using a medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) for 7 days, and then into cochlear hair cell using a medium containing EGF and IGF-1 for the next 14 days. We used the neuroepithelial protein marker nestin and cochlear hair cell marker myosin VIIa as the markers for cells differentiation. Cells expressing the positive markers under the microscope were confirmed to have differentiated into cochlear hair cell. RESULTS: DPSCs were successfully induced to differentiate into NSCs, with mean 24% nestin-positive cells. We found that DPSC-derived NSCs have a great capacity in differentiating into inner ear hair cell-like cells with an average of 81% cells presenting myosin VIIa. Thus, DPSCs have high potential to serve as a good resource for SNHL treatment. CONCLUSION: We found the high potential of DPSCs to differentiate into NSC. The ability of DPSCs in differentiating into neural lineage cell made them a good candidate for regenerative therapy in neural diseases, such as SNHL.

MSCs-released TGFß1 generate CD4+CD25+Foxp3+ in T-reg cells of human SLE PBMC.

BACKGROUND/PURPOSE: Regulatory T-cell (Treg) defects may cause autoreactivity of both T and B cells, leading to autoimmune disease including systemic lupus erythematosus (SLE). The immune response defects in SLE are characterized by the decreased expression of CD4, CD25, and Foxp3, known as inducible Treg (iTreg). Therefore, restoring iTreg expression can reverse autoimmunity states into immune tolerances leading to normal immune responses. Mesenchymal stem cells (MSCs) have immunomodulatory properties to control inflammatory milieu, including in SLE inflammation by releasing TGFß1, IL10, and PGE2, thus MSCs can potentially generate iTreg cells. However, the mechanisms of MSC-released TGFß1 to promote iTreg generation in human SLE remains unclear. This study aims to analyze the role of MSC-released TGFß1 in generating CD4+, CD25+, and Foxp3+ expression in iTreg cells from human SLE peripheral blood mononuclear cells (PBMCs). METHODS: This study used a post-test control group design. MSCs were obtained from human umbilical cord blood and characterized according to their surface antigen expression and multilineage differentiation capacities. PBMCs isolated from SLE patients were divided into five groups, including sham, control, and three treatment groups. The treatment groups were treated by co-culturing MSCs to PBMCs with ratio of 1:1 (T1), 1:25 (T2), and 1:50 (T3) for 72 h incubation. The expression of CD4, CD25, and Foxp3 in Treg was analyzed by flow cytometry assay while TGFß1 level was determined by Cytometric Bead Array (CBA). RESULTS: This study showed that the percentage of CD4+CD25+Foxp3+ iTreg cells was significantly increased in T1 and T2. This finding was aligned with the significant increase of TGFß1 level. CONCLUSION: MSCs promote iTreg cells generation from human SLE PBMCs by releasing TGFß1 to control SLE disease.

The role of mesenchymal stem cells in allergic rhinitis and its relationship with IL-10, plasma cells and regulatory T cells.

Aim Allergic rhinitis (AR) is an IgE-mediated inflammation of the nose. Regulatory T cells (Tregs), plasma cells and inflammatory cytokines have shown to play a critical role in allergic airway inflammation. The aim of the study was to investigate the role of mesenchymal stem cells (MSCs) in generating Treg cells and plasma cells associated with regulating interlukin-10 (IL-10) in AR model. Methods Fifteen male Wistar rats (6 to 8 weeks old) were randomly divided into three groups (control group, sham group, and MSCs treatment group). Ovalbumin (OVA) nasal challenge was conducted daily from day 15 to 21, and MSCs (1x106) were administrated intraperitoneally to OVA-sensitized rats on day 21. Sneezing was observed from day 24 to 27. The rats were sacrificed on day 24 and day 27. The expression of Treg and plasma cells was analysed by flow cytometry assay. The level of IL-10 was analysed under ELISA assay. Results This study showed that the percentage of sneezing and rubbing times significantly decreased in MSCs treatment associated with the regulation of IL-10 level and plasma cell. This finding was aligned with the significant increase of Treg level. Conclusion MSCs administration regulates IL-10 and plasma cell-mediated immune and inflammatory responses while increasing Treg cell production. MSCs may be a promising therapeutic target for treating Treg-mediated allergic diseases.

The effect of honey on post-tonsillectomy pain relief: a randomized clinical trial.

OBJECTIVE: The study aimed to evaluate the effects of honey on the incidence of post-operative pain in patients undergoing tonsillectomy. METHODS: This study is a double-blind and randomized controlled trial design. Twenty-four adult male patients underwent tonsillectomy surgery and were randomized assigned into three groups consist of honey group, placebo group, and control group. All subjects were given standard analgesia and antibiotics, also honey for the honey group and placebo for the placebo group, and only standard post-operative regimens for the control group. This study used silk-cotton tree or kapok tree honey (Ceiba pentandra). Honey was used by gargling every six hours for ten days. Likewise, the same method was applied in the placebo group. Pain scale was assessed for ten days using the Visual Analogue Scale questionnaire, and the frequency of analgesic drugs was recorded on days 1, 2, 4, 7, and 10. RESULT: Honey group showed significantly higher pain reduction when compared to placebo and control groups, with a significant reduction in the pain scale on day 1, 2, 4, 7 and 10 (p = 0.034; p = 0.003; p < 0.001; p = 0.001; p = 0.001) gradually; Significant differences were also observed in analgesic use, especially on day 2, 4 and 7 (p = 0.028; p = 0.001; p = 0.003). CONCLUSIONS: Administration of Kapok tree honey (C. pentandra) after tonsillectomy might reduce post-operative pain and reduce the need for analgesia. Therefore, honey can be considered a complementary medicine and can be administered routinely as adjunctive therapy for post-operative patients.

The Effect of the Intra-Arterial Heparin Flushing (IAHF) Procedure on Hearing Threshold in Sensorineural Hearing Loss Patients.

Background: Sensorineural hearing loss (SNHL) is a frequent problem in Indonesia but its treatment is still limited. This type of hearing loss is related to oxidative stress and decreased vascularization, which can damage the hair cell. The intra-arterial heparin flushing (IAHF) is a procedure that can recover circulation and its agent, namely heparin, also has antioxidant activity. Therefore, the IAHF procedure has the potential to improve hearing function and can be considered an alternative therapy for SNHL. Objective: The study evaluates the effect of the IAHF on hearing improvement based on the difference in hearing threshold values before and after the procedure. Methods: This experimental study used a Pretest-Posttest One-Group Only design. A total of 17 patients with sensorineural hearing loss who met inclusion and exclusion criteria were subjected to pure tone audiometry tests before and 4 hours after the IAHF procedure. The mean difference in hearing threshold was analyzed using paired Students t-test for normally distributed data and Wilcoxon for non-normally distributed data. Results: There was a decrease in the means of hearing threshold in the right and left ear 4 hours after the IAHF procedure. However, based on the paired Students t-test, there was not a significant difference in hearing threshold before and after the procedure (p-value > 0.05). Conclusion: There was hearing threshold improvement 4 hours after the IAHF procedure. This study showed that the IAHF procedure can have a therapeutic effect on sensorineural hearing loss patients.

The Effect of Secreted IL-10 from Mesenchymal Stem Cell on Immune Checkpoint Molecules.

Background: Immunosuppression in sepsis is hypothesized to result from the increased expression of the immune checkpoint molecules programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1). PD-1 and PD-L1 blockade therapies have been reported to increase survival in septic animals. Currently, the interleukin (IL)-10 within mesenchymal stem cell (MSC) secretome is known for its immunomodulatory capacity. Objective: To study the effect of IL-10 within MSC secretome on the expression of immune checkpoints in the rat model of sepsis. Methods: We used 48 male Rattus norvegicus rats in this research and divided them into four groups: sham (rats without sepsis induction and treatment), control (sepsis-induced rats without treatment), T1 (sepsis-induced rats treated with 150 µL of secreted IL-10 from MSC), and T2 (sepsis-induced rats treated with 300 µL of secreted IL-10 from MSC). Forty-eight hours after sepsis induction, we terminated the rats and collected the blood to examine the PD-1 and PD-L1 expression levels. Results: We found a decrease in the relative expression of PD-1 in the septic rat group given 150 µL and 300 µL of secreted IL-10 from MSC compared to the control group, but the decrease was not significant. We also found a decrease in the relative expression of PD-L1 mRNA in the septic rat group given 150 µL and 300 µL of secreted IL-10 from MSC compared to the control group. Conclusion: Administering secreted IL-10 from MSC reduces the expression of PD-1 and PD-L1 in sepsis. These findings suggest that MSC secretome can improve the immunosuppression in sepsis.

The Role of Mesenchymal Stem Cell Secretome in the Inflammatory Mediators and the Survival Rate of Rat Model of Sepsis.

In sepsis, simultaneously elevated levels of pro-inflammatory cytokines and interleukin (IL)-10 indicate immune response dysregulation, increasing the mortality of the host. As mesenchymal stem cell (MSC) secretome is known to have immunomodulatory effects, we aim to assess the role of MSC secretome in the inflammatory mediators (NF-κB p65 and p50, TNF-α, IL-10) and the survival rate of a rat model of sepsis. In this study, forty-eight male Rattus norvegicus rats were divided into one sham group and three groups with sepsis induction: the control group and the sepsis-induced rat groups treated with 150 µL (T1) and 300 µL (T2) of secretome. The survival rate was observed per 6 h for 48 h and plotted using the Kaplan-Meier method. Compared to the control group, T2 showed a significant decrease in the relative expression of NF-κB and the serum TNF-α level, and a significant increase in the serum IL-10 level. Meanwhile, T1 showed a significant decrease in the serum TNF-α level compared to the control group. The Kaplan-Meier Log Rank test did not show significance in the distribution of survival between T1, T2, and the control group. However, from the 18th to the 36th hour, the survival rate of T2 was lower than the survival rate of the control group and T1, with a noticeable difference between T2 and the control group, as well as T1 at the 36th hour. At the 42nd hour, the survival rate of T2 was the same as the control group and remained lower than T1. In conclusion, MSC secretome regulated the inflammatory mediators in rat model of sepsis, with a dose of 150 µL being more effective.

Water Extract Have Superior Cytotoxic Effect Than Ethanolic Extract of Clinacanthus Nutans Leaves in Breast Cancer Stem Cells.

Background: The rapid development of medical technology in managing breast cancer patients still cannot solve the problem of recurrence and resistance. One of the causes of recurrence and molecular resistance is the presence of breast cancer stem cells (BCSCs). Clinacanthus nutans (C.nutans) is a plant found in Medan, North Sumatra, Indonesia. This plant is believed to have anticancer activity in community. Objective: Our study aimed to assess phytochemical of C.nutans leaves, isolate breast cancer stem cells and determine the cytotoxic effects of the ethanolic extract and water extract of C.nutans leaves on breast cancer stem cells at 24, 48, and 72 h of observation. Methods: We underwent the cytotoxic test by using MTT assay and isolated breast cancer stem cells by using MACS and validated them by mammosphere test. Results: We found alkaloids, flavonoids, glycosides and tannins in simplicia and all extracts. BCSCs was valid with the diameter of the mammosphere BCSCs was > 60 µm. The IC50 values of 100%, 60%, 40%, 20% EE, and WE of C.nutans leaves were 227.30; 46.05; 31.12; 98.54, and 16.16 µg/ml respectively in the first 24 hours. In administering WE of C.nutans leaves, BCSCs viability was decreased at 24,48 and 72 hours of observation, namely 69.29±26%; 75.82 ± 21.02% and 38.94±9.34 % (p < 0.0001). Conclusion: The WE of C.nutans leaves had more substantial cytotoxic potential against BCSCs than the EE. The capability of WE C.nutans leaves to suppress BCSC's viability was time-dependent. The anticancer activity were believed originate from alkaloid and flavonoid group.

Correlation Between Intensity of Vimentin Immuno-expression in Young Women with Triple Negative Breast Cancer and Its Cliniocopathological Parameters.

Background: Young breast cancer patients (≤40 years) have different unifying oncogenic signaling pathways when compared to older people. Vimentin is a filament intermediate. Vimentin expression has an important role in epithelial-to-mesenchymal transition (EMT) and cancer progression. Objective: The aim of this study was to assess the correlation between vimentin intensity and clinicopathological parameters in patients with triple negative breast cancer aged 40 years. Methods: Samples were taken from 45 paraffin blocks of patients with young woman with triple negative invasive breast cancer NOS type that met the inclusion criteria, immunohistochemical examination was performed with vimentin. Vimentin intensity was assessed using ImageJ analysis. Results: Samples were taken from paraffin blocks of patients with triple negative invasive breast cancer NOS type that met the inclusion criteria, immunohistochemical examination was performed with vimentin. Vimentin intensity was assessed using ImageJ analysis. Conclusion: This study shows that vimentin intensity is strongly associated with tumor progression, tumor mass size and tumor invasion so that it can be used as a prognostic factor in young triple negative breast cancer patients.

Anti-Aging Activity of Andaliman (Zanthoxylum Acanthopodium DC) Fruit Ethanol Extract on Brain Weight and p16INK4a Expression of Hippocampus in Aging Model Rats.

Background: Physiological aging and due to oxidative stress in long term will have an impact on cellular response disorders, can caused aging of hippocampus and senility. Brain weight is known to decrease with age and p16INK4a as aging biomarkers have been investigated. Andaliman is one of typical herbal plants from North Sumatra has been widely used as an antioxidant, anti-inflammatory and anti-aging. Objective: This study was evaluated effect of andaliman (Zanthoxylum acanthopodium DC) fruit ethanol extract (AEE) on brain weight and p16INK4a expression in aging model rats. Methods: This study was carried out experimentally of 24 male wistar rats. The treatment group consisted of 4 groups; KN= negatif control (normal), KP= positif control (aging model rat), P1 and P2= aging model rat + AEE at dose 150 and 300mg/kgbw, respectively. The aging model rats were D-galactose-induced at dose of 150mg/kgbw for 8 weeks. Brain weigth were recorded by digital scales. p16INK4a expression using immunohistochemical methods. The data analysis with Anova test. Results: This study showed differences brain weight between groups (p=0.523). Brain weight in P1 (1.34±0,06) and P2 (1.30±0.09) tendency increased than KP (1.29±0.62). The p16INK4a expression between groups significant difference (p=0.041), continued with post hoc Least Significant Difference (LSD) showed p16INK4a expression in KN significant decreased than KP (p=0.027). Likewise, p16INK4a expression in P2 was significant decreased than KP (p=0.010). Conclusion: Andaliman ethanol extract at a dose 300mg/kgbw for 8 weeks was improved aging process caused D-galactose induced.

Umbilical Cord-Derived Mesenchymal Stem Cells Improve TGF-ß, α-SMA and Collagen on Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats.

Background: A Erectile dysfunction (ED) is one of the well-known comorbidities in males with diabetes mellitus (DM), whose pathogenesis might be induced by dysregulation of corpus cavernosum smooth muscle cells. UC-MSCs are multipotent cells that attract considerable interest due to immunoregulatory properties and might be a potential strategy to regulate and recover the functional cells and tissues, including tissue improvement in DMED. Objective: This study aims to determine the efficacy of UC-MSCs in improving the erectile function of DMED rats through analyzing the expression of TGF-ß, α-SMA, and collagen. Methods: Total number of 30 male Sprague-Dawley rats (6 to 8 weeks old) were randomly divided into four groups (negative control group, positive control group, T1 group, and T2 group). After 16 h fast, 24 rats were randomly selected and intraperitoneally injected with streptozotocin to induce DM. At 8 weeks after STZ injection, rats with DMED were identified by unresponsive erectile stimulation within 30 min. PC group received 500 µL; T1 rats treated with 500 µL PBS containing 1x106 UC-MSCs; T2 rats treated with 500 µL PBS containing 3x106 UC-MSCs. After MSCs treatment, the rats were sacrificed and the corpus cavernosum tissues were prepared for histological observations. Results: This study resulted in the administration of UC-MSCs could downregulate the expression of TGF-ß, α-SMA, and collagen leading to the improvement of DMED. Conclusion: UC-MSCs improve the expression of TGF-ß, α-SMA, and collagen on erectile dysfunction in streptozotocin-induced diabetic rats.

Revealing the decrease of indoleamine 2,3-dioxygenase as a major constituent for B cells survival post-mesenchymal stem cells co-cultured with peripheral blood mononuclear cell (PBMC) of systemic lupus erythematosus (SLE) patients.

Aim Mesenchymal stem cells (MSCs) have potent immunosuppressive properties to control systemic lupus erythematosus (SLE) disease by inhibiting indoleamine 2,3-dioxygenase (IDO), and increasing regulatory T cells (Treg) to control innate and adaptive immune cells. However, the interaction and mechanism regarding IDO and B cells in the co-culture of MSC and SLE peripheral blood mononuclear cell (PBMCs) remain unclear. This study aimed to investigate the effects of MSCs in controlling B cells through IDO expression in PBMC of SLE patients. Methods This study used a post-test control group design. MSCs were obtained from human umbilical cord blood and characterized according to their surface antigen expression and multilineage differentiation capacities. PBMCs isolated from SLE patients were divided into five groups: sham, control, and three treatment groups. The treatment groups were treated by co-culturing MSCs to PBMCs with a ratio of 1:10, 1:25, and 1:40 for 72 h incubation. The B cell levels were analysed by flow cytometry with cytometric bead array (CBA) and the IDO levels were determined by ELISA. Results The percentages of B cells decreased significantly in groups treated by dose-dependent MSCs, particularly in T1 and T2 groups. These findings were aligned with the significant decrease of the IDO level. Conclusion MSCs control B cells-mediated by a decrease of IDO in PBMC of SLE patients.

Rats' umbilical-cord mesenchymal stem cells ameliorate mast cells and Hsp70 on ovalbumin-induced allergic rhinitis rats.

Aim Allergic rhinitis (AR) is a heterogeneous condition that has been associated with inflammatory responses and is characterized by clinical typical symptoms of nasal itching, sneezing, watery discharge and congestion. Mesenchymal stem cells (MSCs) are multipotent stem cells that have the immunoregulatory ability by secreting various cytokines which potent as a promising therapeutic modality for allergic airway diseases, including AR. The aim of this study was to investigate the effect of rat UC-MSCs on the number of mast cells, the expression of Hsp70 indicated by the nasal symptoms allergic, particularly nasal rubbing in ovalbumininduced AR rats. Methods Fifteen male Wistar rats (6 to 8 weeks old) were randomly divided into three groups (control group, sham group, and OVA+MSCs group). OVA nasal challenge was conducted daily from day 15 to 21, and UC-MSCs (1x106 ) were administrated intraperitoneally to OVA-sensitized rats on day 21. Nasal rubbing was observed from day 22 to 28. The rats were sacrificed on day 22 and day 28. The nasal cavity tissues were prepared for histological observations. Results The administration of UC-MSCs could reduce the number of mast cells and the expression of Hsp70 leading to reduction of nasal symptoms allergic, particularly nasal rubbing. Conclusion Based on this finding, MSCs present a promising immediate curative effect to the inflammatory reaction in AR rats.

The Benefits of Curcuminoid to Expression Nuclear Factor Erythroid 2 Related Factor 2 (NRF2) and Signal to Noise Ratio (SNR) Value in the Noise Exposed Organ of Corti of Rattus Norvegicus.

BACKGROUND: Noise-induced hearing loss (NIHL) can cause damage to the cochlea. Curcumin and nuclear factor erythroid 2-related factor 2 (NRF2) are transcription activators that play a crucial role in defence mechanisms against oxidative stress. The aim of this study was to determine the effect of curcuminoid administration on NRF2 expression, in the organ of Corti of cochlea of Rattus norvegicus that were exposed to noise, from the results of the distortion product otoacoustic emission (DPOAE) examination. METHODS: We divided 36 rats into six groups including Group 1 (control); Group 2 (noise exposure without curcuminoid administration); Group 3 (noise exposure+curcuminoid dose 100 mg/day for four days); Group 4 (noise exposure+curcuminoid dose 200 mg/day for four days); Group 5 (curcuminoid dose of 100 mg/day for 14 days followed by two days of noise exposure); Group 6 (curcuminoid dose 200 mg/day for 14 days followed by two days of noise exposure). RESULTS: Following noise exposure in rats, there was an effect/correlation between NRF2 expression, the SNR values obtained from DPOAE and curcuminoid administration. CONCLUSION: There was a correlation between curcuminoid administration, NRF2 expression and DPOAE treatment. Following noise exposure in rats (Rattus norvegicus), SNR values obtained from DPOAE showed improved cochlear function.

The CD4+CD25+FoxP3+ Regulatory T Cells Regulated by MSCs Suppress Plasma Cells in a Mouse Model of Allergic Rhinitis.

BACKGROUND: Allergic Rhinitis (AR) is the most common immunological disease that has been associated with inflammatory responses and is characterized by sneezing. Previous studies found that AR's allergen exposure significantly induces plasma cells and reduces regulatory T (Treg) cells, a population that contributes to control AR. Therefore, upregulating Treg expression can regulate plasma cells leading to inhibit sneezing in AR. Mesenchymal stem cells (MSCs) are multipotent stem cells that have the immunoregulatory and antiinflammation ability by secreting various cytokines including IL-10 and TGF-ß which potent as a promising therapeutic modality for allergic airway diseases, including AR. OBJECTIVE: To investigate the role of MSCs in generating CD4+, CD25+, and Foxp3+ Regulatory T cells associated with suppressing plasma cell in AR model. METHODS: In this study, fifteen male Wistar rats (6 to 8 weeks old) were randomly divided into three groups (control group, sham group, and MSCs treatment group). OVA nasal challenge was conducted daily from day 15 to 21, and MSCs (1x106) were administrated intraperitoneally to OVA-sensitized rats on day 21. Sneezing was observed from day 22 to 28. The rats were sacrificed on day 22 and day 28. The expression of CD4+ CD25+ Foxp3+ in Treg and plasma cells was analyzed by flow cytometry assay. RESULTS: This study showed that the percentage of plasma cell and sneezing times significantly decreased in MSCs treatment. This finding was aligned with the significant increase of CD4+CD25+Foxp3+ Treg level. CONCLUSION: MSCs administration suppress plasma cells population and sneezing times by up regulating Treg to control AR.

In vitro regulation of IL-6 and TGF-ß by mesenchymal stem cells in systemic lupus erythematosus patients.

Aim To analyse the ability of mesenchymal stem cells (MSCs) to regulate interleukin 6 (IL-6) and transforming growth factor (TGF-ß) expression in vitro under co-culture conditions in human systemic lupus erythematosus (SLE). Method This study used a post-test group design that used peripheral blood mononuclear cells (PBMCs) from SLE patients at Kariadi Hospital, Semarang, Indonesia, and MSCs from a human umbilical cord. The cells were divided into two groups. The control group of PBMCs was treated with a standard medium, and the treatment group was co-cultured with the MSCs at a 1:40 ratio. Following 24 h incubation, the levels of IL-6 and TGF-ß released in the culture medium were measured using a specific ELISA assay. Results This study showed a significant decrease in IL-6 level (p<0.05) and a significant increase in TGF-ß level (p<0.001) following 24 h of co-culture incubation of human SLE PBMCs cells and MSCs. Conclusion The PBMCs-to-MSCs ratio of 1:40 can regulate the IL-6 and TGF-ß levels in human SLE PBMCs.

Effect of Poguntano leaves extract (Picria fel-terrae Merr.) to procalcitonin level in acute bacterial rhinosinusitis model of Wistar mice.

Aim Acute rhinosinusitis (AR) is of viral aetiology and only 0.5- 2% develop into acute bacterial rhinosinusitis. Herbal therapy is a promising alternative in acute bacterial rhinosinusitis treatment. The aim of this study was to evaluate the effect of ethanol extract of Poguntano leaves (EEPL) to procalcitonin level and the amount of bacteria in acute bacterial rhinosinusitis mice model. Methods Experimental research with posttest only control group design in 32 Wistar mice that were divided into 4 groups, 3 of which were being inoculated with Staphylococcus aureus by inserting a sponge to right nasal cavity of the mice (group K2, K3, and K4); another one was the negative control group (K1). Group K2 was not given any kind of therapy (positive control), group K3 was given 10 mg/kd EEPL for 5 days orally during an induction, and group K4 was given 10 mg/kd EEPL for 5 days orally on the 10th day after induction. Mice in the groups K2 and K3 were sacrificed on the 10th day after induction, while mice in group K4 were sacrificed on the 15th day after induction. Result A statistically significant decrease in procalcitonin level (p<0.001) and amount of bacterial colony (p<0.001) was found in four groups. Conclusion Poguntano leaves extract can lower procalcitonin and amount of bacteria colony, showing an anti-inflammatory and antibacterial effect.

Association of endothelial nitric oxide synthase gene (G894T) polymorphism and hypertension in diabetic Bataknese patients.

Aim To assess endothelial nitric oxide synthase (eNOS) gene (G894T) polymorphism and nitric oxide (NO) level in hypertensive diabetic Bataknese patients. Methods A hospital-based, case control study (hypertensive and normotensive diabetic patient) was conducted. Genotyping of eNOS gene (G894T) was done using polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP). Nitric oxide was quantified by sandwich enzyme-linked immunosorbent assay (Sandwich ELISA). Results GT polymorphism and T allele were higher in the hypertensive diabetic patients, 37.5% (p=0.6) and 54% (p=0.39), respectively. Nitric oxide level tended to be lower in the hypertensive diabetic patients (88.87µmol/L) comparing to the normotensive (95.42 µmol/L (p=0.54), as well as GT and TT polymorphism type (p=0.75). Conclusion eNOS gene (G894T) polymorphism is not associated with NO level and hypertension in the diabetic Bataknese patients.

Poguntano effect on cell viability in cholesteatoma keratinocyte of chronic suppurative otitis media.

Aim Chronic Suppurative Otitis Media (CSOM) with or without cholesteatoma is a global major problem and it is becoming a burden especially in developing countries. Studies have found that ethanol extracts of Puguntano leaves obtained by the percolation and socletation methods have phytochemical contents that provide anti-inflammatory effect. The aim of this study was to investigate the effects of ethanol extract and Puguntano on the viability of cholesteatoma keratinocytes. Methods This in vitro experimental study included 8 groups of the cholesteatoma keratinocyte culture: not given puguntano leaf ethanol extract, three positive control groups given different concentrations of dexamethasone, and four groups that were given different concentrations of ethanol extract of leaf Puguntano. Results A statistically significant decrease in procalcitonin level (p<0.001) and an amount of bacterial colonies (p<0.001) in four groups were found. Conclusion This study showed that Puguntano leaf extract has the same effect as dexamethasone in terms of suppressing cell viability and has lower side effects compared to dexamethasone as other herbal medicines. Thus, the ethanol extract of Puguntano leaves can be used as an alternative for the prevention of cholesteatoma.

Hypoxia-Mesenchymal Stem Cells Inhibit Intra-Peritoneal Adhesions Formation by Upregulation of the IL-10 Expression.

BACKGROUND: Intra-peritoneal adhesions (IPAs) common occurre in post abdominal surgical. Athough many methods have been developed for controlling IPAs, including mesenchymal stem cells (MSCs) application, however, there is none completely preventing in due to the mesothelial structure may promote the prolonged inflammations leading. Nevertheless hypoxia-MSCs (H-MSCs) have more potent in controlling the inflammation than normoxia-MSCs (N-MSCs) by releasing several anti-inflamation particularly IL-10, however the H-MSCs application to inhibit IPAs remain unclear. AIM: The aim of this study was to investigate the effectiveness of H-MSCs in preventing the AIPs event by releasing IL-10 on the ileum abrasion sutured omental patch as the animal model of peritoneal adhesion. METHODS: Using 24 IPAs animal model were randomly divided into 4 groups: Sham (Sh), Control (C), H-MSCs at high dose (T1) and H-MSCs at low dose (T2). H-MSCs were incubated under hypoxic conditions (5% O2), 37°C and 5% CO2 for 24 hours. The expression level of IL-10 was performed using RT-PCR analysis. The macroscopic appearance of IPAs was evaluated using Nair's scale base on the absence/presence of adhesion, whereas the microscopic by Zuhlke's scale at Hematoxylin and eosin (H&E) staining. RESULTS: This study showed a significanly increase in IL-10 expression (p < 0.05) at all T groups. In line with this, we also found a significant difference in IPAs between T groups and Control as well as a Sham (p < 0.05) either in the macroscopic or microscopic analysis. CONCLUSION: H-MSCs has a robust ability in inhibiting severe IPAs characterized by the decreased of adhesion formation and the enhanced expression of IL-10.