[Expert consensus on the application of critical care ultrasonography in invasive procedures].

The evolution of critical care medicine is inextricably linked to the development of critical care procedures. These procedures not only facilitate diagnosis and treatment of critically ill patients, but also provide valuable insights into disease pathophysiology. While critical care interventions offer undeniable benefits, the potential for iatrogenic complications necessitates careful consideration. The recent surge in critical care ultrasound (US) utilization is a testament to its unique advantages: non-invasiveness, real-time bedside availability, direct visualization of internal structures, elimination of ionizing radiation exposure, repeatability, and relative ease of learning. Recognizing the need to optimize procedures and minimize complications, critical care utrasound study group of Beijing critical care ultrasound research assocition convened a panel of critical care experts to generate this consensus statement. This document serves as a guide for healthcare providers, aiming to ensure patient safety and best practices in critical care.

[X-ray evaluation of pulp calcification in adult permanent teeth after pulpotomy].

OBJECTIVE: To compare the clinical effects of pulpotomy with two kinds of calcium silicate materials, and to evaluate the formation of dentin bridge and pulp calcification after pulpotomy of adult permanent teeth. METHODS: Patients who visited the General Department of Peking University School and Hospital of Stomatology from November 2017 to September 2019 and planned for pulpotomy on permanent premolars and molars with carious exposed pulp were selected. They were randomly divided into two groups. Bioceramic putty material iRoot BP (iRoot group, n=22) and mineral trioxide aggregate MTA (MTA group, n=21) were used as pulp capping agents, respectively. The patients were recalled after one year and two years. The clinical efficacy, dentin bridge index (DBI) and pulp calcification index (PCI) were recorded. Blinding method was used for the patients and evaluators. RESULTS: There was no significant difference in gender, mean age, dentition and tooth position between the two groups (P>0.05). Seven cases were lost during the first year (4 cases in iRoot group and 3 cases in MTA group). In the iRoot group, 1 case had transient sensitivity at the time of 1-year follow-up. The cure rate of the two groups was 100% at the time of 2-year follow-up. The proportion of dentin bridge formation was 38.9% one year after operation, 55.6% two years after operation. The proportion of partial or even complete disappearance of root canal image was 5.6% before operation, 38.9% and 55.6% one and two years after operation, respectively. The difference was statistically significant by rank sum test (P < 0.05). There was no significant difference in dentin bridge formation and pulp calcification between the two groups (P < 0.05). DBI and PCI after operation was as the same as those before operation (44.4% cases of DBI and 25% cases of PCI) or gradually increased (55.6% cases of DBI and 75% cases of PCI). Spearman's nonparametric correlation analysis showed that age was positively correlated with preoperative pulp calcification index (PCI0, P < 0.05), but not with the dentin bridge index (DBI1, DBI2), pulp calcification index (PCI1, PCI2) and the degree of change (DBI2 vs. DBI1, PCI1 vs. PCI0, PCI2 vs. PCI0) 1-year and 2-year after operation (P>0.05). CONCLUSION: According to this study, good clinical effects were obtained within 2-year after pulpotomy of adult permanent teeth with MTA and iRoot. In some cases, the root canal system had a tendency of calcification aggravation, and there was no statistical difference in the development of this trend between the two groups.

[Evaluation of bioceramic putty repairmen iRoot and mineral trioxide aggregate in mature permanent teeth pulpotomy].

OBJECTIVE: To evaluate the clinical characteristics and effectiveness of pulpotomy in mature permanent teeth with bioceramic putty repairmen iRoot and mineral trioxide aggregate (MTA). METHODS: Pulpotomy was performed on mature permanent premolars and molars with carious exposures at the Department of General Dentistry of Peking University School and Hospital of Stomatology, from November 2017 to September 2019. The patients were randomly divided into 2 groups, Group iRoot (n=22) and Group MTA (n=21). In Group iRoot, bioceramic putty repairmen iRoot was used as pulp capping agent, while in Group MTA, mineral trioxide aggregate was used as pulp capping agent. All the patients had signed informed consent forms. The clinical efficacy was evaluated by clinical examinations (temperature and electrical activity test) and imaging examinations 3, 6, and 12 months after surgery. Blinding was used for the patients and evaluators, but due to the obvious differences in the properties of the two pulp capping agents, the blinding method was not used for the treatment provider (the attending physician). RESULTS: There was no significant difference in gender, average age, dentition and tooth position distribution between the two groups (P>0.05). In the study, 7 cases were lost to follow-up 12 months after operation (4 cases in Group iRoot, and 3 cases in Group MTA). One case in each of the two groups had transient sensitivity at the end of the 3-month follow-up, and the pulp vitality was normal at the end of the 6-month follow-up. One case in Group iRoot showed sensitivity at the end of the 12-month follow-up. The success rates of the two groups at the end of 12-month follow-up were 100%, and the cure rates were 94.4% (Group iRoot) and 100% (Group MTA), respectively, and the difference was not statistically significant (P>0.05). No cases in Group iRoot had obvious crown discoloration, while 3 cases in Group MTA had. CONCLUSION: The clinical characteristics and effectiveness of pulpotomy in mature permanent teeth with bioceramic putty repairmen iRoot were similar with MTA. Bioceramic putty repairmen iRoot is an acceptable material when used in pulpotomy of mature permanent teeth. Because it is not easy to cause tooth discoloration after treatment and is convenient to operate, bioceramic putty repairmen iRoot has a better clinical application prospect.

Gentiana straminea supplementation improves feed intake, nitrogen and energy utilization, and methane emission of Simmental calves in northwest China.

OBJECTIVE: Native plants can be used as additives to replace antibiotics to improve ruminant feed utilization and animal health. An experiment was conducted to evaluate the effects of Gentiana straminea (GS) on nutrient digestibility, methane emissions, and energy metabolism of Simmental calves. METHODS: Thirty-two (5-week-old) male Simmental clves, with initial body weight (BW) of 155±12 kg were fed the same basal diet of concentrates (26%), alfalfa hay (37%), and oat hay (37%) and were randomly separated into four treatment groups according to the amount of GS that was added to their basal diet. The four different groups received different amounts of GS as a supplement to their basal diet during whole experiment: (0 GS) 0 mg/kg BW, the control; (100 GS) 100 mg/kg BW; (200 GS) 200 mg/kg BW; and (300 GS) 300 mg/kg BW. RESULTS: For calves in the 200 GS and 300 GS treatment groups, there was a significant increase in dry matter (DM) intake (p<0.01), average daily gain (ADG) (p<0.05), organic matter intake (p<0.05), DM digestibility (p<0.05), neutral detergent fibre (NDF) digestibility (p<0.05), and acid detergent fibre (ADF) digestibility (p<0.05). Dietary GS supplementation result in quadratic increases of DM intake (p<0.01), ADG (p<0.05), NDF intake (p<0.05), and ADF intake (p<0.05). Supplementing the basal diet with GS significantly increased nitrogen (N) retention (p<0.001) and the ratio of retention N to N intake (p<0.001). Supplementing the basal diet with GS significantly decreased methane (CH4) emissions (p<0.01), CH4/BW0.75 (p<0.05) and CH4 energy (CH4-E) (p<0.05). Dietary GS supplementation result in quadratic increases of CH4 (p<0.01) and CH4/DM intake (p<0.01). Compared with 0 GS, GS-supplemented diets significantly improved their gross energy intake (p<0.05). The metabolizable energy and digestive energy intake were significantly greater for calves in the 100 GS and 200 GS calves than for 0 GS calves (p<0.05). CONCLUSION: From this study, we conclude that supplementing calf diets with GS could improve utilization of feed, energy, and N, and may reduce CH4 emissions without having any negative effects on animal health.

Research Progress on Accumulated Degree Days for PMI Estimation.

ABSTRACT: Among many factors that affect postmortem interval （PMI）, temperature is undoubtedly the most important factor, but simply considering temperature in estimation of PMI has limitations. Accumulated degree days （ADD） is an important indicator related to biological growth and development, which considers two major variables, environmental temperature and elapsed time. Therefore, the establishment of mathematical model of ADD and some evaluation indicators to estimate PMI, is obviously better than single temperature variable. This article briefly describes the research history of ADD, its application in PMI estimation, and the problems it faces, to provide reference for the further development of ADD to improve the accuracy of PMI estimation.

Sequential Changes of Total Adenine Nucleotide and Adenylic-Acid Energy Charge in Muscles of Rats after Death.

ABSTRACT: Objective To determine the purine adenylate [adenosine triphosphate （ATP）, adenosine diphosphate （ADP）, adenosine monophosphate （AMP）] content in the muscles of both hind limbs of rats at different postmortem interval （PMI）, calculate the changes in the total adenine nucleotide （TAN） content and the adenylic-acid energy charge （AEC）, and explore their relationship with PMI. Methods Healthy rats were sacrificed by cervical dislocation and kept at 20 ℃. The muscles of their hind limbs were extracted at 0, 24, 48, 72, 96, 120, 144, and 168 h after death. Reversed-phase high performance liquid chromatography was used to determine the content of purine adenylates, the TAN and AEC of the muscles of the both hind limbs were calculated, and the related regression equations of their relationship with PMI were established. Results Within 168 h of death of rats, the trend of ATP change was different from ADP, and the content of AMP continuously increased. The TAN value gradually increased with the extension of PMI, and the AEC showed a downward trend within 168 h after death. Among them, the patterns of AEC changes with PMI were obvious, the correlation coefficient was high （ R2=0.903）, and the curve fitting relationship was good; the fitting relationship between ATP, ADP, AMP, TAN and PMI was poor （ R2=0.198-0.754）. Conclusion The postmortem change patterns of AEC provide new research ideas for PMI estimation in the forensic field.

[Association between the accumulation of metabolic syndrome abnormal components and arterial pulse wave velocity among adult individuals undergoing routine health examination].

Objective: To explore the association between the accumulation of metabolic syndrome (MS) components and abnormal brachial ankle pulse wave velocity (baPWV), and to investigate the effect of accumulation of abnormal metabolic components on abnormal baPWV among adult individuals undergoing routine health examination. Methods: It's a cross-sectional study. Data from 9 201 stratified sampled subjects, aged between 25 to 75 years old, who took part in the annual health checkups in 11 cities of Zhejiang Province from January to December 2016, were analyzed. Blood pressure(BP), fasting plasma glucose(FPG), waist circumference(WC), triglyceride(TG), high-density lipoprotein cholesterols(HDL-C) were defined as MS components. The baPWV was measured by VP-1000 (BP-203RPE Ⅲ) and ΔbaPWV (measured baPWV-reference baPWV/reference baPWV) ×100 (%)> 10% was defined as abnormal. The relationship between MS components and ΔbaPWV was analyzed by linear regression, and impact of accumulation of MS components on ΔbaPWV was analyzed by logistic regression, and the ΔbaPWV in subjects with different levels of MS components were analyzed by analysis of variance. Results: (1) The linear regression analysis demonstrated a positive correlation between ΔbaPWV and MS components including systolic blood pressure, diastolic blood pressure, WC, FPG, TG, while ΔbaPWV was negatively correlated with HDL-C. The ß values were 0.518, 0.616, 0.208, 2.778, 1.862, -1.339, respectively (all P<0.001), indicating a strong association between ΔbaPWV and systolic blood pressure, diastolic blood pressure, and FPG. (2) The logistic regression analysis showed that the proportion of abnormal ΔbaPWV was 2.595 times higher in MS individuals than in non-MS individuals (P<0.001). Abnormal ΔbaPWV increased in proportion with increase in numbers of the metabolic components (OR=1.913, 2.884, 3.833, 6.161, 11.176 in individuals with 1,2,3,4,5 metabolic components, respectively, all P<0.001). (3) The logistic analysis suggested that each component of MS could affect baPWV,and OR was 4.68, 2.45 for systolic blood pressure and/or diastolic blood pressure and FPG. All combinations of 2 components of MS also significantly affected baPWV,and OR value was the highest (5.104(95%CI 4.281-6.085), P<0.001) for FPG+BP. All combinations of 3 components of MS significantly affected baPWV,and OR value was the highest(5.385(95%CI 4.245-6.831), P<0.001) for BP+FPG+TG. All combinations of 4 components of MS affected baPWV,and OR value was the highest (6.519(95%CI 4.731-8.984), P<0.001) for BP+FPG+WC+TG. (4) Finally, every component of MS was divided into 3 levels, their impact on abnormal ΔbaPWV was analyzed. Prevalence of abnormal ΔbaPWV significantly increased with the increasing levels of the metabolic components expect for HDL-C, the F values were 1 224.66, 832.89, 192.72, 112.79 and 56.22, respectively (all P<0.001), indicating that higher levels of metabolic components significantly affected the ΔbaPWV. Conclusions: MS and accumulation of abnormal MS components are closely related with abnormal ΔbaPWV, and the combination of BP and FPG have the greatest impact on the abnormal ΔbaPWV. Analysis on the accumulation of MS components might serve as an early indicator of arteriosclerosis.

[Technical specification for clinical application of critical ultrasonography].

Critical ultrasonography(CUS) is different from the traditional diagnostic ultrasound, the examiner and interpreter of the image are critical care medicine physicians. The core content of CUS is to evaluate the pathophysiological changes of organs and systems and etiology changes. With the idea of critical care medicine as the soul, it can integrate the above information and clinical information, bedside real-time diagnosis and titration treatment, and evaluate the therapeutic effect so as to improve the outcome. CUS is a traditional technique which is applied as a new application method. The consensus of experts on critical ultrasonography in China released in 2016 put forward consensus suggestions on the concept, implementation and application of CUS. It should be further emphasized that the accurate and objective assessment and implementation of CUS requires the standardization of ultrasound image acquisition and the need to establish a CUS procedure. At the same time, the standardized training for CUS accepted by critical care medicine physicians requires the application of technical specifications, and the establishment of technical specifications is the basis for the quality control and continuous improvement of CUS. Chinese Critical Ultrasound Study Group and Critical Hemodynamic Therapy Collabration Group, based on the rich experience of clinical practice in critical care and research, combined with the essence of CUS, to learn the traditional ultrasonic essence, established the clinical application technical specifications of CUS, including in five parts: basic view and relevant indicators to obtain in CUS; basic norms for viscera organ assessment and special assessment; standardized processes and systematic inspection programs; examples of CUS applications; CUS training and the application of qualification certification. The establishment of applied technology standard is helpful for standardized training and clinical correct implementation. It is helpful for clinical evaluation and correct guidance treatment, and is also helpful for quality control and continuous improvement of CUS application.

Optically pumped lasing with a Q-factor exceeding 6000 from wet-etched GaN micro-pyramids.

We report the observation of room-temperature optically pumped lasing modes from a single GaN pyramid microcavity on a metallic mirror. The mode at 367.2 nm exhibits a low threshold (0.4-0.5 MW/cm2) and a narrow linewidth (0.054 nm), by which the quality factor can be estimated to be >6000. These lasing behaviors can be attributed to the specific wet-etching approach by selectively etching away defects and pyramid geometry with bottom Ag reflectors for better light confinement. Optical resonances in these pyramids are further investigated in combination with three-dimensional finite-difference time-domain simulations.

Gleason sum upgrading between biopsy and radical prostatectomy in Chinese population: Updated nomograms. / Infragradación de la biopsia de próstata respecto a la pieza de prostatectomía la población china: nomogramas actualizados.

INTRODUCTION: To assess the risk factors of Gleason sum upgrading between biopsy and radical prostatectomy (RP) and update the nomogram for the prediction of Gleason sum upgrading. METHODS: The study cohort consisted of 237 Chinese prostate adenocarcinoma patients who underwent 10-core prostate biopsy and subsequently received RP in Huashan Hospital from February 2011 to May 2015. The main outcome of our study was Gleason sum upgrading between biopsy and RP pathology. Univariate and multivariate logistic regression models were conducted to explore the potential predictors, and ultimately to build the nomograms. The prediction model was further evaluated for its ability to predict significant upgrading in patients with biopsy Gleason sum<8. RESULTS: In the main cohort of all the patients, Gleason sum upgrading was observed in 62 (26.16%) patients. The pre-operative prostate-specific antigen (PSA) level, biopsy Gleason sum, and digital rectal examination were used in building the nomogram, which was validated internally with a bootstrap-corrected concordance index of 0.787. In the sub-cohort of 115 patients with standardized biopsy details, Gleason sum upgrading was observed in 31 (26.96%) patients. The pre-operative PSA level, biopsy Gleason sum, and number of positive cores were used in the nomogram, which was also validated internally with a bootstrap-corrected concordance index of 0.833. These two nomograms both demonstrated satisfactory statistical performance for predicting significant upgrading. CONCLUSIONS: Updated nomograms to predict Gleason sum upgrading in Chinese population between biopsy and RP were developed, demonstrating good statistical performance upon internal validation.

Evaluation of the clinical significance of classification of traumatic anterior shoulder instability using double-contrast computed tomography arthrography.

This study evaluated the clinical significance of traumatic anterior shoulder instability (TASI) classification using double-contrast computed tomography (CT) arthrography. Patient were randomly assigned to two groups: group 1 (n = 62); and group 2 (n = 63). TASI symptom severity in group 1 was assessed using physical signs of shoulder trauma and conventional X-ray, CT and magnetic resonance imaging; these patients received either conservative management (with physical rehabilitation) or standard surgery. Group 2 underwent double-contrast CT arthrography to classify TASI; its findings formed the basis of subsequent management. At 24 months post-therapy, significant improvements in clinical outcomes were observed in group 2: Constant scores were higher and Western Ontario Shoulder Instability Index scores were lower. At 24 months, recurrence rates were 21.0% (13/62) in group 1 and 7.9% (5/63) in group 2. Findings suggested that TASI classification using double-contrast CT arthrography provided meaningful information thereby improving treatment efficacy.

Vascular regression and survival are differentially regulated by MT1-MMP and TIMPs in the aortic ring model of angiogenesis.

This study was designed to investigate the role of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) in the reabsorption of neovessels in collagen gel cultures of rat and mouse aortic rings. Aortic angiogenesis was associated with collagen lysis and production of the matrix-degrading enzymes MMP-2, MMP-9, and membrane-type MMP (MT1-MMP, or MMP-14). Vascular growth and regression were not affected by disruption of MMP-2 or MMP-9. In addition, no effect on vascular regression was observed by blocking plasmin, a protease implicated in the activation of MMPs, with epsilon-aminocaproic acid or by adding plasminogen, which caused a modest increase in vascular proliferation. Conversely, angiogenesis was blocked and vessels stabilized by inhibiting MT1-MMP with neutralizing antibodies, TIMP-2, TIMP-3, or TIMP-4. TIMP-1, which blocks MMP-2 and MMP-9 but is a poor inhibitor of MT1-MMP, had no antiangiogenic effect. However, TIMP-1 prolonged the survival of neovessels following angiogenesis. Vascular regression was accelerated in aortic cultures from TIMP-1- and TIMP-2-deficient mice. The vascular survival effect of anti-MT1-MMP antibodies and TIMPs with MT1-MMP inhibitory activity was associated with complete inhibition of collagen lysis. In contrast, TIMP-1 had no anticollagenolytic effect. These results indicate that MT1-MMP plays a critical role not only in angiogenesis but also in vascular regression and demonstrate that TIMPs with anti-MT1-MMP activity have opposite effects on angiogenic outcomes depending on the stage of the angiogenic process. This study also suggests the existence of a TIMP-1-mediated alternate pathway of vascular survival that is unrelated to MT1-MMP inhibitory activity.

A putative mechanism on remission of multiple sclerosis during pregnancy: estrogen-induced indoleamine 2,3-dioxygenase by dendritic cells.

The basis for the reduced relapse rate of multiple sclerosis (MS) during pregnancy remains unexplained but, if defined, could create novel treatment options. Estrogen constitutes one candidate molecule, but the mechanism by which estrogen may affect MS during pregnancy is unclear. In this study, we used monocyte-derived dendritic cells (DCs) from MS patients to explore the estrogen (17-beta-estradiol)-related pathway of immune modulation. Estrogen induced the expression of indoleamine 2,3-dioxygenase (IDO) on DCs, limiting T-cell proliferation and both Th1 and Th2 cytokine production. The suppression of T-cell proliferation mediated by estrogen-exposed DCs was partly abolished by the IDO-inhibitor, 1-methyl-dl-tryptophan, indicating that estrogen-exposed DCs induced IDO-dependent T-cell suppression. Our data support the hypothesis that the change in the clinical course of MS observed in pregnancy may be related to the estrogen-DC-IDO axis, which could represent a novel target for MS therapy.

Use of vascular explants for ex vivo neovascularization of biomaterials.

Biomaterial polymers have been proposed as scaffolds for cell assembly in vascular bioengineering. We describe here a new method for the neovascularization of polyurethane meshes from explants of rat aorta. Aortic rings embedded in collagen-permeated polyurethane meshes and cultured in medium supplemented with fetal bovine serum and vascular endothelial growth factor generated florid microvascular outgrowths that efficiently vascularized the available spaces between polyurethane fibers. The neovessels could be identified in the live cultures by phase-contrast microscopy, and in formalin-fixed preparations by the ABC peroxidase procedure, using the endothelial-specific Griffonia isolectin B4. The aortic outgrowths were successfully labeled with the intravital fluorescent dyes Calcein AM or SPDiOC(18), which are nontoxic and can be used for tracking studies. This study shows that artificial biomaterial meshes can be colonized ex vivo with histotypic microvascular networks, and provides the proof of concept for the future development of stably vascularized devices for in vivo implantation.

Effects of stress factors, bioregulators, and synthetic precursors on indole alkaloid production in compact callus clusters cultures of Catharanthus roseus.

Compact callus cluster (CCC) cultures established from Catharanthus roseus consist of cohesive callus aggregates displaying certain levels of cellular or tissue differentiation. CCC cultures synthesize about two-fold more indole alkaloids than normal dispersed-cell cultures. Our studies here show that additions of KCl, mannitol, and a variety of synthetic precursors and bioregulators to the CCC cultures markedly improved indole alkaloid production and release of these alkaloids into the medium. Treatment with 250 mM mannitol and 4 g/l KCl yielded 42.3 mg l(-1) and 33.6 mg l(-1)of ajmalicine, respectively; these amounts were about four-fold higher than the control. Succinic acid, tryptamine, and tryptophan feedings also significantly increased ajmalicine (41.5 mg l(-1), 36.9 mg l(-1), and 31.8 mg l(-1), respectively) and catharanthine (21.1 mg l(-1), 17.2 mg l(-1), and 18 mg l(-1), respectively) production by the CCC cultures, while geraniol feeding inhibited biomass and alkaloid accumulation. We also found that tetramethyl ammonium bromide could significantly improve ajmalicine production (49.3 mg l(-1)) and catharanthine production (18.3 mg l(-1)) in C. roseus CCC cultures. The mechanisms responsible for these treatment effects are discussed herein.

Selection of fungal elicitors to increase indole alkaloid accumulation in catharanthus roseus suspension cell culture.

Various fungal elicitors derived from 12 fungi were tested to improve indole alkaloid production in Catharanthus roseus cell suspension cultures. Results show that different fungal mycelium homogenates stimulate different kinds of indole alkaloid (ajmalicine, serpentine and catharanthine) accumulation, which ranged from 2- to 5-fold higher than the control. Some fungal culture filtrates also efficiently elicited the biosynthesis of different indole alkaloids. The optimal elicitor addition and exposure time for the maximal alkaloid production were on day 7 after subculture and for 3 days of treatment but different fungal elicitors showed the different optimal treatment dosages. Additions of elicitor at the doses ranging from 5 mg/l to 30 mg/l of carbon hydrate equivalent resulted in varieous amounts and kinds of indole alkaloid accumulation. Exposed to a same fungal elicitor, several different cell lines generated the different responses regarding as growth rate, culture color and alkaloid production.

Enhanced catharanthine production in catharanthus roseus cell cultures by combined elicitor treatment in shake flasks and bioreactors.

Chemical and fungal elicitors were added to Catharanthus roseus cell suspension cultures so as to improve the production of indole alkaloids. A synergistic effect on alkaloid accumulation was observed in C. roseus cell cultures when treated with some combined elicitors of fungal preparations and chemicals. Among them, the combination of tetramethyl amminium bromide and Aspergillum niger mycelial homogenate gave the highest ajmalicine yield (63 mg l(-1)) and an improved catharanthine accumulation (17 mg l(-1)). The combined elicitors of malate and sodium alginate resulted in the highest catharanthine yield (26 mg l(-1)) and a high ajmalicine accumulation (41 mg l(-1)) in the cell cultures. Based on the synergistic effect of malate and sodium alginate, a process with enhanced catharanthine production in Catharanthus roseus cell cultures was developed in shake flasks and a bioreactor. After 10 days of culture, 25 mg l(-1), 32 mg l(-1) and 22 mg l(-1) catharanthine yield were obtained in 500-ml flasks, 1000-ml flasks and in a 20-l airlift bioreactor, respectively. Upon malate-alginate combining treatments, peroxidase, catalase and superoxide dismutase activities decreased in elicited cells but phenylalanine ammonia lyase and lipoxygenase activities increased dramatically. That suggests a typical defense responses took place in the combined elicitors-treated cell cultures. Furthermore, the combined elicitors also caused a significant increase of malondialdehyde level in cell cultures, which suggests a serious lipid peroxidation occurred in the elicited cell cultures. Comparison of these results suggests that malate and alginate combining treatment also stimulates defense responses, such as lipid peroxidation, in all C. roseus culture processes and this may mediate the indole alkaloid production via jasmonate pathway.