The optimization conditions of establishing an H9c2 cardiomyocyte hypoxia/reoxygenation injury model based on an AnaeroPack System.

Ischemia-reperfusion (I/R) injury is a major cause of cardiomyocyte apoptosis after vascular recanalization, which was mimicked by a hypoxia/reoxygenation (H/R) injury model of cardiomyocytes in vitro. In this study, we explored an optimal H/R duration procedure using the AnaeroPack System. To study the H/R procedure, cardiomyocytes were exposed to the AnaeroPack System with sugar and serum-free medium, followed by reoxygenation under normal conditions. Cell injury was detected through lactate dehydrogenase (LDH) and cardiac troponin (c-Tn) release, morphological changes, cell apoptosis, and expression of apoptosis-related proteins. The results showed that the damage to H9c2 cells increased with prolonged hypoxia time, as demonstrated by increased apoptosis rate, LDH and c-Tn release, HIF-1α expression, as well as decreased expression of Bcl-2. Furthermore, hypoxia for 10 h and reoxygenation for 6 h exhibited the highest apoptosis rate and damage and cytokine release; in addition, cells were deformed, small, and visibly round. After 12 h of hypoxia, the majority of the cells were dead. Taken together, this study showed that subjecting H9c2 cells to the AnaeroPack System for 10 h and reoxygenation for 6 h can achieve a practicable and repeatable H/R injury model.

MBD2-mediated Th17 differentiation in severe asthma is associated with impaired SOCS3 expression.

T helper 17 (Th17) cells has proven to be crucial in the pathogenesis of severe asthma. Although it is known that Suppressor of cytokine signaling 3 (SOCS3) is involved in differentiation of Th17 cells but, how it affects severe asthma is uncertain. Since previous studies indicated that Methtyl-CpG binding domain protein 2 (MBD2) null mice was deficient in Th17 cell differentiation, the aim of the present study was to understand how MBD2 interacts with SOCS3 to regulate Th17 cell differentiation in severe asthma. Here, we show that SOCS3 expression was significantly decreased in Th17-mediated severe asthmatic mice, accompanied by elevated STAT3 phosphorylation and RORγt expression. Knock-down of SOCS3 promoted the differentiation of naïve T cells into Th17 cells through STAT3/RORγt pathway. Meanwhile, MBD2 was overexpressed in Th17-mediated severe asthmatic mice. Intervention of MBD2 expression lead to a negative change of SOCS3 expression, whereas the differentiation of Th17 cells showed positive change. In addition, MBD2 knockout (MBD2-KO) mice displayed increased SOCS3 expression and decreased Th17 differentiation after severe asthma modeling. Taken together, our results suggest that MBD2 might facilitate Th17 cell differentiation via down-regulating SOCS3 expression in severe asthma. These findings uncover new roles for SOCS3 and MBD2, and provide a potential target for treatment of severe asthma.

MBD2 Regulates Th17 Cell Differentiation and Experimental Severe Asthma by Affecting IRF4 Expression.

Th17 cells and IL-17 participate in airway neutrophil infiltration characteristics in the pathogenesis of severe asthma. Methyl-CpG binding domain protein 2 (MBD2) expression increased in CD4+ T cells in peripheral blood samples of asthma patients. However, little is known about that epigenetic regulation of MBD2 in both immunological pathogenesis of experimental severe asthma and CD4+ T cell differentiation. Here, we established a neutrophil-predominant severe asthma model, which was characterized by airway hyperresponsiveness (AHR), BALF neutrophil granulocyte (NEU) increase, higher NEU and IL-17 protein levels, and more Th17 cell differentiation. In the model, MBD2 and IRF4 protein expression increased in the lung and spleen cells. Under overexpression or silencing of the MBD2 and IRF4 gene, the differentiation of Th17 cells and IL-17 secretion showed positive changes. IRF4 protein expression showed a positive change with overexpression or silencing of the MBD2 gene, whereas there was no significant difference in the expression of MBD2 under overexpression or silencing of the IRF4 gene. These data provide novel insights into epigenetic regulation of severe asthma.

Recent advances of traditional Chinese medicine against cardiovascular disease: overview and potential mechanisms.

Cardiovascular disease (CVD) is the leading cause of human mortality worldwide. Despite Western medicine having made encouraging results in the clinical management of CVD, the morbidity, mortality, and disability rates of the disease remain high. Modern pharmacology has confirmed that traditional Chinese medicine (TCM), characterized by its multi-component, multi-target, and integrity, plays a positive and important role in the prevention and treatment of various CVDs in China, which has notable advantages in stabilizing disease, improving heart function, and enhancing the quality of life. Importantly, TCM is gradually being accepted by the international community due to its low cost, high safety, versatile bioactivity, and low toxicity. Unfortunately, comprehensive studies on the therapeutic effect of TCM on CVD and its mechanisms are very limited, which may restrict the clinical application of TCM in CVD. Therefore, this review is performed to analyze the pathogenesis of CVD, including inflammatory response, oxidative stress, mitochondrial dysfunction, pyroptosis, ferroptosis, dysbiosis of gut microbiota, etc. Moreover, we summarized the latest progress of TCM (formulas, extracts, and compounds) in curing CVD according to published literature from 2018 to 2023, as well as its mechanisms and clinical evidence. In conclusion, this review is expected to provide useful information and reference for the clinical application of TCM in the prevention and treatment of CVD and further drug development of CVD.

A study of pharmacists-joint total parenteral nutrition in haematopoietic cell transplantation in accord with diagnosis related groups: A retrospective clinical research.

Background: Within Diagnosis Related Groups, based on service capability, efficiency, and quality safety assessment, clinical pharmacists contribute to promoting rational drug utilisation in healthcare institutions. However, a deficiency of pharmacist involvement has been observed in the total parenteral nutrition support to patients following haematopoietic cell transplantation (HCT) within DRGs. Methods: This study involved 146 patients who underwent HCT at the Department of Haematology, the Second Affiliated Hospital of Dalian Medical University, spanning from January 2020 to December 2022. Results: Patients were allocated equally, with 73 in the control group and 73 in the pharmacist-involved group: baseline characteristics showed no statistics significance, including age, body mass index, nutrition risk screening-2002 score, liver and kidney function, etc. Albumin levels, prealbumin levels were significantly improved after a 7-day TPN support (34.92 ± 4.24 vs 36.25 ± 3.65, P = 0.044; 251.30 ± 95.72 vs 284.73 ± 83.15, P = 0.026). The body weight was increased after a 7-day support and before discharge (58.77 ± 12.47 vs 63.82 ± 11.70, P = 0.013; 57.61 ± 11.85 vs 64.92 ± 11.71, P < 0.001). The length of hospital stay, costs and the rate of re-admissions were significantly shortened (51.10 ± 1.42 vs 46.41 ± 1.86, P = 0.048; 360,162.67 ± 91,831.34 vs 324,070.16 ± 112,315.51, P = 0.035; 61.64% vs 43.84%, P = 0.046). Conclusions: Pharmacist-joint TPN support enhances the service efficiency score of medical units, ensuring the fulfilment of orders and rational medication.

How Rhizosphere Microbial Assemblage Is Influenced by Dragon Fruits with White and Red Flesh.

The synthesis of betalain using microorganisms is an innovative developmental technology, and the excavation of microorganisms closely related to betalain can provide certain theoretical and technical support to this technology. In this study, the characteristics of soil microbial community structures and their functions in the rhizospheres of white-fleshed dragon fruit (Hylocereus undatus) and red-fleshed dragon fruit (Hylocereus polyrhizus) were analyzed. The results show that the soil bacterial and fungal compositions in the rhizospheres were shaped differently between H. undatus and H. polyrhizus. Bacterial genera such as Kribbella and TM7a were the unique dominant soil bacterial genera in the rhizospheres of H. undatus, whereas Bradyrhizobium was the unique dominant soil bacterial genus in the rhizospheres of H. polyrhizus. Additionally, Myrothecium was the unique dominant soil fungal genus in the rhizospheres of H. polyrhizus, whereas Apiotrichum and Arachniotus were the unique dominant soil fungal genera in the rhizospheres of H. undatus. Moreover, TM7a, Novibacillus, Cupriavidus, Mesorhizobium, Trechispora, Madurella, Cercophora, and Polyschema were significantly enriched in the rhizospheres of H. undatus, whereas Penicillium, Blastobotrys, Phialemonium, Marasmius, and Pseudogymnoascus were significantly enriched in the rhizospheres of H. polyrhizus. Furthermore, the relative abundances of Ascomycota and Penicillium were significantly higher in the rhizospheres of H. polyrhizus than in those of H. undatus.

Targeted reversal of multidrug resistance in ovarian cancer cells using exosome­encapsulated tetramethylpyrazine.

The objective of the present study was to develop exosomes (EXOs) encapsulating tetramethylpyrazine (TMP) for the reversal of drug resistance in ovarian cancer therapy. Human A2780 cells were incubated with TMP for 48 h. Purified TMP­primed EXOs (EXOs­TMP) were isolated through ultracentrifugation. The developed EXOs­TMP were characterized using techniques such as transmission electron microscopy, nanoparticle tracking analysis, Fluorescence microscopy and western blotting. Subsequently, MTT, western blotting and flow cytometry assays were performed to evaluate the biological effects in drug­resistant A2780T cells. The results demonstrated that the incorporation of TMP into EXOs exhibited an anti­ovarian cancer effect and markedly enhanced the antitumor efficacy of paclitaxel (PTX). Furthermore, it was identified that the ability of EXO­TMP to reverse cell resistance was associated with the downregulation of multidrug resistance protein 1, multidrug resistant­associated protein 1 and glutathione S­transferase Pi protein expression. Flow cytometry analysis revealed that EXO­TMP induced apoptosis in drug­resistant cells and enhanced the apoptotic effect when combined with PTX. EXOs are naturally sourced, exhibit excellent biocompatibility and enable precise drug delivery to target sites, thereby reducing toxic side effects. Overall, EXO­TMP exhibited direct targeting capabilities towards A2780T cells and effectively reduced their drug resistance. EXOs­TMP provide a novel and effective drug delivery pathway for reversing drug resistance in ovarian cancer.

The Role of Multifidus in the Biomechanics of Lumbar Spine: A Musculoskeletal Modeling Study.

BACKGROUND: The role of multifidus in the biomechanics of lumbar spine remained unclear. PURPOSE: This study aimed to investigate the role of multifidus in the modeling of lumbar spine and the influence of asymmetric multifidus atrophy on the biomechanics of lumbar spine. METHODS: This study considered five different multifidus conditions in the trunk musculoskeletal models: group 1 (with entire multifidus), group 2 (without multifidus), group 3 (multifidus with half of maximum isometric force), group 4 (asymmetric multifidus atrophy on L5/S1 level), and group 5 (asymmetric multifidus atrophy on L4/L5 level). In order to test how different multifidus situations would affect the lumbar spine, four trunk flexional angles (0°, 30°, 60°, and 90°) were simulated. The calculation of muscle activation and muscle force was done using static optimization function in OpenSim. Then, joint reaction forces of L5/S1 and L4/L5 levels were calculated and compared among the groups. RESULTS: The models without multifidus had the highest normalized compressive forces on the L4/L5 level in trunk flexion tasks. In extreme cases produced by group 2 models, the normalized compressive forces on L4/L5 level were 444% (30° flexion), 568% (60° flexion), and 576% (90° flexion) of upper body weight, which were 1.82 times, 1.63 times, and 1.13 times as large as the values computed by the corresponding models in group 1. In 90° flexion, the success rate of simulation in group 2 was 49.6%, followed by group 3 (84.4%), group 4 (89.6%), group 5 (92.8%), and group 1 (92.8%). CONCLUSIONS: The results demonstrate that incorporating multifidus in the musculoskeletal model is important for increasing the success rate of simulation and decreasing the incidence of overestimation of compressive load on the lumbar spine. Asymmetric multifidus atrophy has negligible effect on the lower lumbar spine in the trunk flexion posture. The results highlighted the fine-tuning ability of multifidus in equilibrating the loads on the lower back and the necessity of incorporating multifidus in trunk musculoskeletal modeling.

Increased Proportion of Dual-Positive Th2-Th17 Cells Promotes a More Severe Subtype of Asthma.

Asthma is a heterogeneous disease, and abnormal activation of T cells is the driving link of asthma's pathophysiological changes. Dual-positive Th2-Th17 cells, as newly discovered T-helper cells, have the functions of Th2 and Th17 cells and can coproduce Th2 and Th17 cytokines. Previous studies have shown that dual-positive Th2-Th17 cells increase the chances of asthma and correlate with asthma severity. However, the exact role of dual-positive Th2-Th17 cells in asthma is not known. Since there is no mature differentiation method for dual-positive Th2-Th17 cells, the present study aimed to clarify the strict differentiation conditions and reveal how dual-positive Th2-Th17 cells regulate asthma phenotypes. In this study, we confirmed that IL-1ß, IL-6, anti-IFN-γ, and IL-21 promoted biphenotypic cell differentiation. Moreover, more proportion of dual-positive Th2-Th17 cells can be obtained by conditioned differentiation of mouse CD4+ T cells after classical allergic asthma modeling. Before asthma modeling, adoptive dual-positive Th2-Th17 cells promoted T cells to differentiate into the same biphenotype cells and exacerbated the severity of asthma. Together, our results clarify the differentiation conditions of dual-positive Th2-Th17 cells and further confirm that it stimulates asthma T cells to differentiate into the same biphenotype cells, leading to exacerbation of asthma.

Nano Porous Carbon Derived from Citrus Pomace for the Separation and Purification of PMFs in Citrus Processing Wastes.

The by-product of citrus juice processing is a huge source of bioactive compounds, especially polymethoxyflavones (PMFs) and fibers. In this study, a method for the separation and purification of PMFs from citrus pomace was established based on citrus nanoporous carbon (CNPC) enrichment. Different biomass porous carbons were synthesized, their adsorption/desorption characteristics were evaluated, and the CNPCs from the peel of Citrus tangerina Tanaka were found to be best for the enrichment of PMFs from the crude extracts of citrus pomace. Using this method, six PMF compounds including low-abundant PMFs in citrus fruits such as 5,6,7,4'-tetramethoxyflavone and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone can be simultaneously obtained, and the purities of these compounds were all higher than 95%, with the highest purity of nobiletin reaching 99.96%. Therefore, CNPCs have a great potential for the separation and purification of PMFs in citrus processing wastes, potentially improving the added value of citrus wastes. We also provide a method reference for disposing of citrus pomace in the future.

MicroRNA-16 inhibits the proliferation and metastasis of human lung cancer cells by modulating the expression of YAP1.

PURPOSE: Accounting for significant human morbidity and mortality across the globe, lung cancer is the most prevalent type of cancer as far as incidence and mortality is concerned. MicroRNAs (miRs) have shown an amazing potential to act as therapeutic agents for the management of several human diseases. This study investigated the function of miR-16 in lung cancer. METHODS: The normal lung cancer cell line MRC3 and lung cancer cell lines SK-MES-1, A549, MS-53 and SK-LU-1 were used in the present study. The qRT-PCR was used for expression profiling of miR-16 and yes associated protein 1 (YAP1). WST-1 assay was used to monitor the proliferation rate. Flow cytometry was used for cell cycle analysis. Apoptosis was examined by DAPI and annexin V/propidium iodide (PI) staining. TargetScan analysis was performed to identify the potential target of miR-16 and western blot analysis was done to estimate the protein expression. RESULTS: The gene expression analysis showed miR-16 to be suppressed in lung cancer tissues and cell lines. Overexpression of miR-16 inhibited the growth and metastasis of the DMS-53 lung cancer cells via induction of the apoptotic cell death. Bioinformatic approaches revealed miR-16 exerts its effects by targeting YAP1. YAPI expression was found elevated in lung cancer tissues and its silencing halted the growth of the DMS-53 lung cancer cells. Nonetheless, YAP1 overexpression could reverse the growth inhibitory effects of miR-16. CONCLUSION: Taken together, miR-16 may serve as novel therapeutic target for the treatment of lung cancer.

Bronchial epithelial cells of young and old mice directly regulate the differentiation of Th2 and Th17.

To determine whether or not house dust mite (HDM) and HDM+lipopolysaccharide (LPS) exposure causes a difference in T-cell subsets from young and old mice. The bronchial epithelial cells (BECs) from young and old mice were divided into three groups (PBS (control), HDM, and HDM+LPS). CD4+ naive T cells from the spleen and lymph nodes were collected after 24 h of co-culture with BECs. The number of Th2 and Th17 cells was elevated in the HDM and HDM+LPS groups compared with the control group; these responses were exacerbated when exposed to HDM+LPS. The number of HDM- and HDM+LPS-specific Th2/Th17 cells in young mice was higher than old mice; however, the Th2:Th17 cell ratio was greater in young mice, whereas the Th17:Th2 cell ratio was greater in old mice. The expression of GATA-3 and RORc was increased in the HDM+LPS and HDM groups compared with the PBS group and exhibited most in HDM+LPS group. The expression of HDM+LPS-specific GATA-3 in young mice was higher, while the expression of HDM+LPS-specific RORc in old mice was higher. Murine BECs directly regulated CD4+ naive T-cell differentiation under allergen exposure.

Advances in Application of Mechanical Stimuli in Bioreactors for Cartilage Tissue Engineering.

Articular cartilage (AC) is the weight-bearing tissue in diarthroses. It lacks the capacity for self-healing once there are injuries or diseases due to its avascularity. With the development of tissue engineering, repairing cartilage defects through transplantation of engineered cartilage that closely matches properties of native cartilage has become a new option for curing cartilage diseases. The main hurdle for clinical application of engineered cartilage is how to develop functional cartilage constructs for mass production in a credible way. Recently, impressive hyaline cartilage that may have the potential to provide capabilities for treating large cartilage lesions in the future has been produced in laboratories. The key to functional cartilage construction in vitro is to identify appropriate mechanical stimuli. First, they should ensure the function of metabolism because mechanical stimuli play the role of blood vessels in the metabolism of AC, for example, acquiring nutrition and removing wastes. Second, they should mimic the movement of synovial joints and produce phenotypically correct tissues to achieve the adaptive development between the micro- and macrostructure and function. In this article, we divide mechanical stimuli into three types according to forces transmitted by different media in bioreactors, namely forces transmitted through the liquid medium, solid medium, or other media, then we review and summarize the research status of bioreactors for cartilage tissue engineering (CTE), mainly focusing on the effects of diverse mechanical stimuli on engineered cartilage. Based on current researches, there are several motion patterns in knee joints; but compression, tension, shear, fluid shear, or hydrostatic pressure each only partially reflects the mechanical condition in vivo. In this study, we propose that rolling-sliding-compression load consists of various stimuli that will represent better mechanical environment in CTE. In addition, engineers often ignore the importance of biochemical factors to the growth and development of engineered cartilage. In our point of view, only by fully considering synergistic effects of mechanical and biochemical factors can we find appropriate culture conditions for functional cartilage constructs. Once again, rolling-sliding-compression load under appropriate biochemical conditions may be conductive to realize the adaptive development between the structure and function of engineered cartilage in vitro.

Novel amphiphilic folic acid-cholesterol-chitosan micelles for paclitaxel delivery.

In order to decrease the toxicity of paclitaxel (PTX) and increase the efficiency, we developed an amphiphilic PTX injection system using a biodegradable and biocompatible polymer synthesized by folic acid, cholesterol, and chitosan (FACC). This FACC-based polymer had a low critical concentration (64.13µg/ml) and could self-assemble in aqueous condition to form nanoscale micelles. The particle sizes of FACC-PTX micelles were 253.2±0.56 nm, the encapsulation efficiency and loading capacity of these FACC-PTX micelles were 65.1±0.23% and 9.1±0.16%, respectively. The cumulative release rate was about 85% at pH 5.0 which was higher than that at pH 7.4 (76%). This pH-dependent release behavior was highly suggesting that PTX release from FACC-PTX micelles might be higher in a weak acidic tumor microenvironment and lower toxic for normal cells. The anti-cancer effectiveness of FACC-PTX micelles was investigated by in vitro cytotoxicity and targeting study. The results revealed that FACC micelles have non-toxic on cells as evidenced by high cell viability found (86% to 100%) in the cells cultured with various concentrations of FACC micelles (1 to 500 µg/ml). Targeting study indicated that the cytotoxic efficacy of FACC-PTX micelles was significantly higher than that with Taxol® in the Hela cells (folate receptor-positive cells). These findings indicated that the anticancer efficiency of PTX can be enhanced by adding some cancer cell positive receptor into drug carrier and the FACC micelle was a potential tumor targeting carrier for PXT delivery.

A novel dual-frequency loading system for studying mechanobiology of load-bearing tissue.

In mechanobiological research, an appropriate loading system is an essential tool to mimic mechanical signals in a native environment. To achieve this goal, we have developed a novel loading system capable of applying dual-frequency loading including both a low-frequency high-amplitude loading and a high-frequency low-amplitude loading, according to the mechanical conditions experienced by bone and articular cartilage tissues. The low-frequency high-amplitude loading embodies the main force from muscular contractions and/or reaction forces while the high-frequency low-amplitude loading represents an assistant force from small muscles, ligaments and/or other tissue in order to maintain body posture during human activities. Therefore, such dual frequency loading system may reflect the natural characteristics of complex mechanical load on bone or articular cartilage than the single frequency loading often applied during current mechanobiological experiments. The dual-frequency loading system is validated by experimental tests using precision miniature plane-mirror interferometers. The dual-frequency loading results in significantly more solute transport in articular cartilage than that of the low-frequency high-amplitude loading regiment alone, as determined by quantitative fluorescence microscopy of tracer distribution in articular cartilage. Thus, the loading system can provide a new method to mimic mechanical environment in bone and cartilage, thereby revealing the in vivo mechanisms of mechanosensation, mechanotransduction and mass-transport, and improving mechanical conditioning of cartilage and/or bone constructs for tissue engineering.

Evaluation of a risk factor scoring model in screening for undiagnosed diabetes in China population.

OBJECTIVE: To develop a risk scoring model for screening for undiagnosed type 2 diabetes in Chinese population. METHODS: A total of 5348 subjects from two districts of Jinan City, Shandong Province, China were enrolled. Group A (2985) included individuals from east of the city and Group B (2363) from west of the city. Screening questionnaires and a standard oral glucose tolerance test (OGTT) were completed by all subjects. Based on the stepwise logistic regression analysis of Group A, variables were selected to establish the risk scoring model. The validity and effectiveness of this model were evaluated in Group B. RESULTS: Based on stepwise logistic regression analysis performed with data of Group A, variables including age, body mass index (BMI), waist-to-hip ratio (WHR), systolic pressure, diastolic pressure, heart rate, family history of diabetes, and history of high glucose were accepted into the risk scoring model. The risk for having diabetes increased along with aggregate scores. When Youden index was closest to 1, the optimal cutoff value was set up at 51. At this point, the diabetes risk scoring model could identify diabetes patients with a sensitivity of 83.3% and a specificity of 66.5%, making the positive predictive value 12.83% and negative predictive value 98.53%. We compared our model with the Finnish and Danish model and concluded that our model has superior validity in Chinese population. CONCLUSIONS: Our diabetes risk scoring model has satisfactory sensitivity and specificity for identifying undiagnosed diabetes in our population, which might be a simple and practical tool suitable for massive diabetes screening.

Appropriate insulin initiation dosage for insulin-naive type 2 diabetes outpatients receiving insulin monotherapy or in combination with metformin and/or pioglitazone.

BACKGROUND: Few studies have given suggestions on appropriate initiation insulin dosage when combined with oral antidiabetic drugs (OADs). This research was to investigate appropriate initiation insulin doses for insulin-naive type 2 diabetes patients with different combinations and the relationship between insulin dosage and relevant factors. METHODS: This was a randomized, open-label, treat to target study. The target was 20% decrease of both fasting plasma glucose (FPG) and 2 hours post-breakfast blood glucose (P2hBG). One hundred and forty-seven insulin-naive Chinese patients recruited were randomly assigned to 3 groups: group A, patients received insulin monotherapy; group B, received insulin plus metformin (0.5 g, tid) and group C, received insulin plus metformin (0.5 g, tid) and pioglitazone (15 mg, qd). Insulin doses were initiated with a dose of 0.3 U×kg(-1)×d(-1) and titrated according to FPG and P2hBG till reached the targets. RESULTS: Both the time of getting 20% reduction of FPG and P2hBG showed significant differences among the three groups. The time was shortest in Group C. The insulin doses needed to achieve glucose reduction of 20% in three treatment groups were (0.40 ± 0.04) U×kg(-1)×d(-1) for Group A, (0.37 ± 0.04) U×kg(-1)×d(-1) for Group B, and (0.35 ± 0.03) U×kg(-1)×d(-1) for Group C, respectively. Multiple linear stepwise regression analysis showed that insulin doses correlated with body weight, FPG, diabetes duration, age and history of sulfonylurea treatment. The standardized regression coefficients were 0.871, 0.322, 0.089, 0.067 and 0.063 (with all P < 0.05). CONCLUSIONS: To achieve blood glucose's reduction of 20% within safety context, initial insulin doses were recommended as the following: 0.40 U×kg(-1)×d(-1) for insulin mono-therapy, 0.37 U×kg(-1)×d(-1) for insulin plus metformin treatment, and 0.35 U×kg(-1)×d(-1) for insulin plus metformin and pioglitazone treatment in Chinese type 2 diabetes outpatients. Body weight is found the most closely related factor to the insulin dosage.

[Association of coxsackie virus infection and T lymphocyte subset changes with type 1 diabetes].

OBJECTIVE: To investigate the relationship between coxsackievirus infection and type 1 diabetes mellitus (T1DM), and observe the changes of T lymphocyte subsets in the development of T1DM. METHODS: We detected Coxsackievirus RNA by reverse transcription PCR, and measured the change in T-lymphocyte subsets by flow cytometry in 22 cases of newly diagnosed T1DM (group I), 30 patients with diabetes for some time (group II), and 30 healthy subjects (group III). RESULTS: The positivity rate of coxsackie virus RNA in groups I, II, and III was 55.55%, 23.33%, and 6.67%, respectively, showing a significant difference among the 3 groups (P<0.01). Patients with upper respiratory tract infection had a higher positivity rate for coxsackie virus RNA than those without upper respiratory tract infection in group I (P<0.05). Compared with the control group, the percentage of CD3, CD4 and CD4/CD8 ratio decreased significantly in groups I and II (P<0.01 or P<0.05). CD3, CD4 and CD4/CD8 tended to increase in group II in comparison with group I, and there was an significant difference in CD3 and CD4 between the two groups (P<0.01 or P<0.05). Compared with the control group and CVBRNA-negative group, CVBRNA-positive group showed significantly lowered CD3, CD4, CD8 and CD4/CD8 (P<0.01 or P<0.05). CONCLUSION: The occurrence and development of type 1 diabetes is closely related to coxsackie virus infection, and the changes in T lymphocyte subsets serves as a probable mechanism of its pathogenicity.

Protective effects of Salvia plebeia compound homoplantaginin on hepatocyte injury.

Salvia plebeia R. Br is a traditional Chinese herb which has been considered as an inflammatory mediator used for treatment of many infectious diseases including hepatitis. Previously, the compound homoplantaginin was isolated in our group. Hence, we evaluated the protective effects of homoplantaginin on hepatocyte injury. Homoplantaginin displayed an antioxidant property in a cell-free system and showed IC(50) of reduction level of DPPH radical at 0.35 microg/ml. In human hepatocyte HL-7702 cells exposed to H(2)O(2), the addition of 0.1-100 microg/ml of homoplantaginin, which did not have a toxic effect on cell viability, significantly reduced lactate dehydrogenase (LDH) leakage, and increased glutathione (GSH), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in supernatant. In vivo assay, we employed the model of Bacillus Calmette-Guérin (BCG)/lipopolysaccharide (LPS)-induced hepatic injury mice to evaluate efficacy of homoplantaginin. Homoplantaginin (25-100mg/kg) significantly reduced the increase in serum alanine aminotranseferase (ALT) and aspartate aminotransferase (AST), decreased the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1). The same treatment also reduced the content of thiobarbituric acid-reactive substances (TBARS), elevated the levels of GSH, GSH-Px and SOD in hepatic homogenate. The histopathological analysis showed that the grade of liver injury was ameliorated with reduction of inflammatory cells and necrosis of liver cells in homoplantaginin treatment mice. These results suggest that homoplantaginin has a protective and therapeutic effect on hepatocyte injury, which might be associated with its antioxidant properties.