[Mediating effect of unhealthy lifestyle and depressive symptom on association between life course factors and ageing health].

Objective: To explore the mediating effect of unhealthy lifestyle and depressive symptom on the associations between life course factors and aging health. Methods: The study included 6 217 participants (aged ≥45 years) from the China Health and Retirement Longitudinal Study (CHARLS). We used principal component analysis (PCA) and hierarchical clustering analysis (HCA) to divide participants into six subgroups based on 70 life course factors. Five key life course factors were identified based on correlation analysis and their contribution to aging health. Physiological dysregulation (PD) was calculated by using eight biomarkers in the 2015 CHARLS biomarker dataset. Linear regression, logistic regression, and mediation models were used to explore the complex associations of life course subgroups, key factors, unhealthy lifestyle, depression symptom with PD. Results: Life course subgroups were significantly associated with PD after adjusting chronological age and gender (ß: 0.08-0.17, all P<0.05). Life-course subgroups and key factors, including adverse experiences in adulthood and lower education level, were significantly associated with unhealthy lifestyle (ß: 0.04-0.52, all P<0.05). Life-course subgroups and key factors, including childhood trauma, parental health in childhood, adverse experiences in adulthood, and lower education level, were significantly associated with depression symptom (OR: 1.16-4.76, all P<0.05). Mediation analysis showed that unhealthy lifestyle had partial mediating effect on the association of life course subgroups and key factors, including adverse experiences in adulthood, and lower education levels, with PD (3.1%-3.6%). Depression symptom had partial mediating effect on the association of life course subgroups and key factors, including childhood trauma, adverse experience in adulthood, and lower education level, with PD (6.0%-16.2%). Conclusions: Unhealthy lifestyle and depression symptom has partial mediating effect on the impact of life course factors on aging health. It is important to pay attention to these two modifiable factors while targeting childhood trauma and adverse experience in adulthood.

[Naringenin inhibits thoracic aortic aneurysm formation in mice with Marfan syndrome].

OBJECTIVE: To identify whether naringenin plays a protective role during thoracic aneurysm formation in Marfan syndrome. METHODS: To validate the effect of naringenin, Fbn1C1039G/+ mice, the mouse model of Marfan syndrome, were fed with naringenin, and the disease progress was evaluated. The molecular mechanism of naringenin was further investigated via in vitro studies, such as bioluminescence resonance energy transfer (BRET), atomic force microscope and radioligand receptor binding assay. RESULTS: Six-week-old Fbn1C1039G/+ mice were fed with naringenin for 20 weeks. Compared with the control group, naringenin significantly suppressed the aortic expansion [Fbn1C1039G/+ vs. Fbn1C1039G/++naringenin: (2.49±0.47) mm, n=18 vs. (1.87±0.19) mm, n=22, P < 0.05], the degradation of elastin, and the expression and activity of matrix metalloproteinase 2 (MMP2) and MMP9 in the ascending aorta of Fbn1C1039G/+ mice. Besides, treatment with naringenin for 6 weeks also attenuated the disease progress among the 20-week-old Fbn1C1039G/+ mice with established thoracic aortic aneurysms [Fbn1C1039G/+ vs. Fbn1C1039G/++naringenin: (2.24±0.23) mm, n=8 vs. (1.90±0.17) mm, n=8, P < 0.05]. To understand the underlying molecular mechanisms, we examined the effects of naringenin on angiotensin Ⅱ type 1 receptor (AT1) signaling and transforming growth factor-ß (TGF-ß) signaling respectively, which were the dominant signaling pathways contributing to aortopathy in Marfan syndrome as previously reported. The results showed that naringenin decreased angiotensin Ⅱ (Ang Ⅱ)-induced phosphorylation of protein kinase C (PKC) and extracellular regulating kinase 1/2 (ERK1/2) in HEK293A cell overexpressing AT1 receptor. Moreover, naringenin inhibited Ang Ⅱ-induced calcium mobilization and uclear factor of activated T-cells (NFAT) signaling. The internalization of AT1 receptor and its binding to ß-arrestin-2 with Ang Ⅱ induction were also suppressed by naringenin. As evidenced by atomic force microscope and radioligand receptor binding assay, naringenin inhibited Ang Ⅱ binding to AT1 receptor. In terms of TGF-ß signaling, we found that feeding the mice with naringenin decreased the phosphorylation of Smad2 and ERK1/2 as well as the expression of TGF-ß downstream genes. Besides, the serum level of TGF-ß was also decreased by naringenin in the Fbn1C1039G/+ mice. Furthermore, we detected the effect of naringenin on platelet, a rich source of TGF-ß, both in vivo and in vitro. And we found that naringenin markedly decreased the TGF-ß level by inhibiting the activation of platelet. CONCLUSION: Our study showed that naringenin has a protective effect on thoracic aortic aneurysm formation in Marfan syndrome by suppressing both AT1 and TGF-ß signaling.

Effect of one bout of local vibration exercise with blood flow restriction on neuromuscular and hormonal responses.

This study aimed to investigate the effects of single local vibration (LV) with and without blood flow restriction (BFR) on muscle activity and hormonal responses. A total of 12 physically inactive males were exposed to 10 sets of intermittent LV (35-40 Hz) on unilateral mid-quadriceps in the supine lying position and LV + BFR (inflated to 140 mmHg) sessions in a repeated-measures randomized crossover design, with a 1-week interval separating the sessions. The results indicated that the electromyography values from the rectus femoris during LV + BFR were greater than those during LV (p < 0.05). LV + BFR caused a minor increase in the lactate (LA) response (p < 0.05); LV with or without BFR failed to elicit change in growth hormone (GH) and testosterone (T) levels (p > 0.05). Cortisol (C) levels were decreased postexercise in both the sessions (p < 0.05). In conclusion, BFR elicited higher increase in muscle activity and metabolic response, but it did not induce hormonal responses. The exposure of LV and LV + BFR may only have a relief effect as detected by the reduction in C levels, probably because the LV did not elicit sufficient stimulus to the muscles.

The role of miR-146b-5p in TLR4 pathway of glomerular mesangial cells with lupus nephritis.

OBJECTIVE: MicroRNAs (miRNAs) are now recognized as important regulators of gene expression. The aim of the study was to investigate the role of miR-146b-5p in the TLR4 pathway and provide the basis for the treatment of lupus nephritis. MATERIALS AND METHODS: The glomerular mesangial cells were cultured in vitro and divided into 3 groups: control group, a group of miR-146b-5p mimic added, and a group of miR-146b-5p inhibitor added. The levels of IL-6 and IL-8 in the cell culture supernatant of the three groups were detected by ELISA. The cell proliferation was detected by MTT. The expressions of MiR-146b-5p and TLR4 pathway-associated factor TRAF6 were detected by RT- PCR. The expression of TRAF6 and IRAK1 protein was detected by Western blot. RESULTS: The overexpression of miR-146b-5p could reduce the level of IL6 and IL8 in cell culture and inhibit glomerular mesangial cell proliferation in some degree. Also, the overexpression of miR-146b-5p could inhibit the expressions of TLR4 pathway-associated factor TRAF6 and IRAK1mRNA, and the expressions of TRAF6 and IRAK1 protein. CONCLUSIONS: MiR-146b-5p attenuated the inflammatory response of glomerular mesangial cells by inhibiting the expressions levels of TRAF6 and IRAK1 in lupus nephritis.

NAC attenuates adriamycin-induced nephrotic syndrome in rats through regulating TLR4 signaling pathway.

OBJECTIVE: Nephrotic syndrome (NS) is a detrimental renal disease that affects a large population. It is suggested that Toll-like Receptor 4 (TLR4) signaling pathway plays an important role in NS. The aim of this study was to evaluate the immunosuppressive effect of N-acetylcysteine (NAC) in the treatment of NS elucidate its interaction with TLR4 pathway in a rat model. MATERIALS AND METHODS: Rat NS model was constructed using the Bertain method by injecting adriamycin (4.5 mg/kg) intravenously at day 1, and injecting 2 mg/kg adriamycin (ADR) at day 7. NS rats were treatment with NAC of 150 mg/kg daily through gavage. Control rats received equivalent amounts of saline daily. Quantitative Real-time PCR was used to evaluate TLR4 expression in kidney tissues after treatments. Western blot analysis was used to evaluate NF-κBp65 expression. ELISA was used to evaluate the expression of immunological factors, including TNF-α, IL-6, and IL-1ß. RESULTS: Rat NS models demonstrated higher protein levels in urine, accompanied by an increased in the TLR4 level. After NAC treatment, TLR4 level was reduced. NAC treatment also attenuated the NF-κBp65 overexpression in NS rats. Concomitantly, TNF-α, IL-6, and IL-1ß levels, which are indicators of immunological and informatory responses, were also decreased after NAC treatment. CONCLUSIONS: NAC treatment ameliorated nephrotic syndrome in NS rat models by suppressing TLR4 signaling, as well as immunological and inflammatory responses.

Pachymic acid ameliorates sepsis-induced acute kidney injury by suppressing inflammation and activating the Nrf2/HO-1 pathway in rats.

OBJECTIVE: To investigate the protective effects and underlying mechanisms of pachymic acid (PA) on sepsis-induced acute kidney injury (AKI). MATERIALS AND METHODS: Sepsis-induced AKI model was made by cecal ligation and puncture (CLP) surgery in SD rats. Animals were randomly divided into 5 groups: a sham group, a CLP group, and three PA-treated groups, which received intraperitoneal injection of PA at the dosage of 5, 20 and 50 mg/kg.bw, respectively. Kidney index, Cre and BUN contents were determined to evaluate the renal function. Pathological changes of kidney tissue were observed by HE staining. Levels of inflammatory mediators (TNF-α, IL-6) were measured to assess the inflammation in renal tissue. Moreover, the expression levels of iNOS, Nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were studied by Real-time PCR and Western blot. RESULTS: PA treatment can significantly decrease the kidney index, and notably drop the contents of Cre and BUN. Renal pathological damage was also found to be effectively improved by PA in a dose-dependent manner. PA treatment was observed to inhibit the renal inflammation by reducing the TNF-α and IL-6 levels. Besides, PA treatment significantly decreased the expression levels of iNOS, and enhanced the expression of Nrf2 and HO-1 in the kidney. CONCLUSIONS: PA had potential therapeutic effects on sepsis-induced AKI in rats, and the activity may be associated with the anti-inflammatory function and antioxidant effect via activating Nrf2/ HO-1 pathway.

The value of the bulbocavernosus reflex and pudendal nerve somatosensory evoked potentials in distinguishing between multiple system atrophy and Parkinson's disease at an early stage.

OBJECTIVES: This study was designed to investigate the clinical value of the bulbocavernosus reflex (BCR) and pudendal nerve somatosensory evoked potentials (PSEPs) in the differential diagnosis between multiple system atrophy (MSA) and Parkinson's disease (PD) in early stage. MATERIALS AND METHODS: A total of 31 patients with MSA, 45 patients with PD, and 60 healthy participants were included in this study. A Keypoint EMG/EP system was used for BCR and PSEP measurements. Electrophysiological parameters were collected for statistical analysis. RESULTS: The BCR elicitation rates were significantly lower in the patients with MSA than in the patients with PD (P<.05). Prolonged BCR latencies were found in the MSA group compared to the PD and control groups (P<.05). Bulbocavernosus reflex latencies were significantly prolonged in patients with MSA compared with PD patients showing early urogenital symptoms (P<.05). There was no significant difference in PSEP P41 latencies among the three groups (P=.434 in males, P=.948 in females). Both BCR and PSEP amplitudes were significantly lower in the MSA/PD group than in the control group (P<.001). CONCLUSIONS: Pudendal nerve damage is more severe in MSA than in PD. Prolonged BCR latency may be valuable for distinguishing between MSA and PD in the early stages. BCR and PSEP testing may also contribute to localized and qualitative diagnosis of the distribution of neurodegenerative pathologies in these two disorders.

Development of polymorphic microsatellite markers based on expressed sequence tags in Populus cathayana (Salicaceae).

Populus cathayana occupies a large area within the northern, central, and southwestern regions of China, and is considered to be an important reforestation species in western China. In order to investigate the population genetic structure of this species, 10 polymorphic microsatellite loci were identified based on expressed sequence tags from de novo sequencing on the Illumina HiSeq 2000 platform. All microsatellite primers were tested on 48 P. cathayana individuals from four locations on the Qinghai-Tibet Plateau. The observed heterozygosity ranged from 0.000 to 1.000, and the null-allele frequency ranged from 0.000 to 0.904. These microsatellite markers may be a useful tool in genetic studies on P. cathayana and closely related species.

[Effects of Rad9 mutants with impaired DNA mismatch repair function on tumorigenesis of colorectal cancer].

OBJECTIVE: The aim of this study was to investigate the effects of Rad9 mutants with impaired DNA mismatch repair (MMR) function on the tumorigenesis of colorectal cancer. METHODS: The colorectal cancer tumor samples were collected from 100 patients. The mutation profiles of human Rad9 (hRad9) gene in these samples were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing. The plasmid of pFLAG-hRad9 (L101M) was constructed following the QuickChange mutagenesis procedure and transfected into mRad9-deleted mouse cells (mRad9(-/-) cells). The expression of hRad9 protein was measured by western blot analysis. The MMR activity in live cells was detected by flow cytometry using the reporter plasmid for MMR function. RESULTS: Mutation from Leu to Met at the residue 101 (L101M) of hRad9 gene was detected in 7 of the 100 samples. The mismatch repair efficiency of mRad9(-/-)+ L101M cells (mRad9-deleted mouse cells with ectopic expression of L101M hRad9 gene) was (34.0±5.6)%, which was significantly lower than that in the mRad9(-/-)+ hRad9 cells [mRad9-deleted mouse cells with ectopic expression of hRad9 gene, (48.0±7.5)%, P<0.05]. After N-nitroso-N-methylurea (MNU) treatment, the survival rate of mRad9(-/-)+ L101M cells was (33.7±5.9)%, which was significantly higher than that in the mRad9(-/-)+ hRad9 cells [(21.3±4.7)%, P<0.05]. Thus, ectopic expression of L101M hRad9 gene resulted in significantly reduced MMR activity and increased resistance to MNU. Furthermore, ectopic expression of hRad9 gene with mutation at the target residues of post-translational modification in mRad9(-/-) cells also led to a reduced MMR activity. CONCLUSION: Rad9 mutants with impaired DNA mismatch repair function may promote tumorigenesis of colorectal cancer.

Changes in astrocyte functional markers and ß-amyloid metabolism-related proteins in the early stages of hypercholesterolemia.

Cholesterol is an essential substance for maintaining normal structure and function of the brain. But unfortunately, a long-term high-cholesterol diet can lead to a variety of pathological changes of the brain such as ß-amyloid (Aß) accumulation, Tau hyperphosphorylation, reactive gliosis, neuroinflammation, neuronal death and synaptic degeneration. These pathological changes have complex internal relations with one other, causing memory impairment and participating in the pathogenesis of Alzheimer's disease (AD). However, early hypercholesterolemia-induced events that lead to brain deterioration are not clear. To address this, 6-month-old female mice were fed a 3% cholesterol diet for 8weeks, followed by behavioral, biochemical and neuropathological analyses. The high-cholesterol-fed mice did not show neuronal and synaptic impairment or cognitive deficits compared with mice given a normal diet, but astrocytes were mildly activated with increased expression of functional markers including apolipoprotein E and aquaporin 4 in the hippocampus. Hippocampal interleukin-1ß expression slightly increased, but interleukin-6 (IL-6) and tumor necrosis factor-α did not change significantly compared with those in the control group. Levels of Aß, and its precursor protein, were unaffected, but levels of presenilin 1 and insulin-degrading enzyme (IDE), that initiate Aß generation and degradation, respectively, increased in the hippocampus of the model mice. In addition, Tau phosphorylation levels were not different between the control and model groups. These results suggest that changes in astrocyte functional markers and Aß metabolism proteins, which contribute to maintaining brain cholesterol and Aß homeostasis, are early events in the process of hypercholesterolemia-related neuropathological changes.

A normobaric gas exposure animal chamber and its validation in hyperoxia exposure.

We are living in an environment full of gases, and any change in the concentration of a component of the air or contaminants (usually toxic) in the air may significantly threaten human health. Thus, to investigate the influence of gases in animal models it is helpful to elucidate the pathogenesis of gas-related injury. Although there are devices used for gas exposure in animals, there are still limitations in the establishment of these animal models, such as the change in gas concentration during the refreshing of water, food and litter, and the contamination of toxic gases released by animals. Herein, we freshly prepared a chamber for normobaric gas exposure. During the exposure in this chamber, the refreshing of water, food and litter does not require opening of the chamber. The chamber gases are continuously circulated and filtered, and the gas concentration remains very stable. To validate the feasibility of this chamber, rats were exposed to pure oxygen as an example. Results showed that rats with hyperoxia-induced lung injury simulated by pure oxygen exposure displayed the representative characteristics as observed in humans: shortness of breath, lung edema, alveolar septal rupture, infiltration of inflammatory cells, oxidative and inflammatory injury. This suggests that it is feasible to establish animal models using this chamber for the investigation of gas toxicity.

First Report of Alternaria alternata Causing Black Leaf Spot of Rubber Tree in China.

We first reported Alternaria heveae (E.G. Simmons ) to be the pathogen that caused black leaf spot of rubber tree (Hevea brasiliensis Muell. Arg) in Heikou county in July 2014 (1). Black leaf spots that resembled the symptoms caused by A. heveae were observed on the leaves of rubber trees of the whole propagule collection nursery in Jingping County (22°68' N and 103°05' E) of Yunnan Province. Black foliar spots (0.1 to 2 mm in diameter) surrounded by a yellow halo with lesions slightly sunken on the leaf surface were observed. To confirm whether the disease was caused by the same pathogen, 5-mm2 sections were removed from the leading edge of the lesion and were surface-sterilized in 75% ethanol, air-dried, plated on potato carrot agar (PCA), and incubated at 28°C in the dark. Colonies of the fungus on PCA had round margins and little aerial mycelia with gray-black coloration after 6 days of growth on PCA (2). Medium brown conidia were found to be in short chains of two to eight spores, ovoid, obclavate, and obpyriform, with or without a short conical or cylindrical-shaped apical beak. Conidia ranged from 22.5 to 67.5 µm long (mean 39.9 µm) × 10 to 15 µm wide (mean 12.5 µm; 100 colodia were measured), with three to six transverse septa and zero to three longitudinal or oblique septa. Morphological characteristics matched the descriptions of A. alternata [(Fries) Keissler] (4).The ITS1-5.8S-ITS2 region of one single-spore isolate, Ah02JP1, was amplified with primers ITS1 and ITS4. The PCR product was sequenced directly and deposited in GenBank (Accession No. KM111289). A BLAST search of the GenBank database revealed 100% similarity with A. alternata isolates KJ829535.1, KJ677246.1, and KF813070.1. Therefore, the pathogen was identified as A. alternata on the basis of its morphological characteristics and ITS sequence. Pathogenicity of a representative isolate, Ah02JP1 was confirmed using a field rubber tree inoculation method. Three rubber plants (the clone of rubber tree Yunyan77-4) were grown to the copper-colored leaf stage. Leaves were spray-inoculated (104 conidia per milliliter spore suspension) until drops were equally distributed using a manual pressure sprayer. Three rubber plants sprayed with sterile distilled water were used as controls. After inoculation, the plants were covered with plastic bags to maintain high relative humidity. The plastic bags were removed 2 days post-inoculation (dpi), and the plants were monitored daily for symptom development. Five days post-inoculation, spots similar to the original ones seen on the field trees developed on all inoculated leaves, while control leaves remained symptomless. A. alternata was re-isolated from spray-inoculated leaves, confirming Koch's postulates. A. alternata has been reported as the causal agent of leaf blight of rubber tree in India, which initially appeared as minute spots on leaves and enlarged with the growth of the leaves (3). However, in the present study, the symptoms (black leaf spots) remained small over time after inoculation. To our knowledge, this is the first report of A. alternata on rubber tree in China. Correct identification of pathogens is essential for disease management strategies. This report will establish a foundation for the further study of Alternaria alternata to address the disease effectively. References: (1) Z. Y. Cai et al. Plant Dis. 98:1011, 2014. (2) E. Mirkova. J. Phytopathol. 151:323, 2003. (3) C. B. Roy et al. J. Plantation Crops 34:499, 2006. (4) T. Y. Zhang. Page 32 in: Flora Fungorum Sinicorum, Vol. 16: Alternaria. Science Press, Beijing, 2003.

First Report of Alternaria heveae Causing Black Leaf Spot of Rubber Tree in China.

Rubber tree (Hevea brasiliensis Muell. Arg.) is an important industrial crop of tropical areas for natural rubber production. In October 2013, foliar spots (0.1 to 0.4 mm in diameter), black surrounded by a yellow halo, and with lesions slightly sunken were observed on the rubber tree leaf in a growing area in Heikou County of Yunnan Province. Lesion tissues removed from the border between symptomatic and healthy tissue were surface sterilized in 75% ethanol and air-dried, plated on PDA plates, and incubated at 28°C with alternating day/night cycles of light. The pathogen was observed growing out of many of the leaf pieces, and produced abundant conidia. Colonies 6.1 cm in diameter developed on potato carrot agar (PCA) after 7 days, with well-defined concentric rings of growth. Colonies on PCA were composed of fine, dark, radiating, surface and subsurface hyphae. Conidia produced in PCA culture were mostly solitary or in short chains of 2 to 5 spores, long ovoid to clavate, and light brown, 40 to 81.25 × 8 to 20 µm (200 colonies were measured), with 3 to 6 transverse septa and 0 to 2 longitudinal or oblique septa. Morphological characteristics were similar to those described for Alternaria heveae (3,4). A disease of rubber tree caused by Alternaria sp. had been reported in Mexico in 1947 (2). DNA of Ah01HK13 isolate was extracted for PCR and sequencing of the ITS region with ITS1 and ITS4 primers was completed. From the BLAST analysis, the sequence of Ah01HK13 (GenBank Accession No. KF953884), had 97% similarity to A. dauci, 96% identical to A. macrospora (AY154701.1 and DQ156342.1, respectively), indicating the pathogen belonged to Alternaria genus. According to morphological characteristics, this pathogen was identified as A. heveae. Pathogenicity of representative isolate, Ah01HK13 was confirmed using a field rubber tree inoculation method. Three rubber plants (the clone of rubber tree Yunyan77-4) were grown to the copper-colored leaf stage and inoculated by spraying spore suspension (concentration = 104 conidia/ml) to the copper-colored leaves until drops were equally distributed on it using manual pressure sprayer. Three rubber plants sprayed with sterile distilled water were used as controls. After inoculation, the plants were covered with plastic bags. The plastic bags were removed after 2 days post-inoculation (dpi) and monitored daily for symptom development (1). The experiment was repeated three times. The typical 0.1 to 0.4 mm black leaf spots were observed 7 dpi. No symptoms were observed on control plants. A fungus with the same colony and conidial morphology as A. heveae were re-isolated from leaf lesions on inoculated rubber plants, but not from asymptomatic leaves of control plants, fulfilling Koch's postulates. Based on these results, the disease was identified as black spot of rubber tree caused by A. heveae. To our knowledge, this is the first report of A. heveae on rubber tree in China. References: (1) Z. Y. Cai et al. Microbiol Res. 168:340, 2013. (2) W. J. Martin. Plant Dis. Rep. 31:155, 1947. (3) E. G. Simmons. Mycotaxon 50:262, 1994. (4) T. Y. Zhang. Page 111 in: Flora Fungorum Sinicorum: Alternaria, Science Press, Beijing, 2003.

Comparison of methylation procedures for conjugated linoleic acid and artifact formation by commercial (trimethylsilyl) diazomethane.

Four different methods for methylating conjugated linoleic acid (CLA) were compared. The HCl/MeOH and BF(3)/MeOH methods were tested under different time and temperature combinations. Increasing temperature and/or incubation time for either method decreased the cis-9,trans-11 and trans-10,cis-12 isomers, but trans-9,trans-11/trans-10,trans-12 isomers and artifacts (allylic methoxide) were increased. In addition, the triacylglyceride form of CLA was tested using the above methods and NaOMe at various temperatures for 20 min. The NaOMe did not generate methoxy artifacts. However, there were impurities in GC after methylation with NaOMe as well as with BF(3)/MeOH. The (trimethylsilyl)diazomethane method, which is a mild and easy alternative, was tested. Free forms of fatty acids were easily, but not completely, methylated by this method. Also, the method generated artifacts (trimethylsilyl CLA esters) and impurities (trimethylsilyl) that would interfere with short-chain fatty-acid analysis by GC.

Huperzine-A in capsules and tablets for treating patients with Alzheimer disease.

AIM: To compare the efficacy and safety between huperzine-A (Hup) in capsules and tablets for treating patients with Alzheimer disease (AD). METHODS: Using multicenter, prospective, double-blind, double-mimic, parallel, positive controlled and randomized methods, 60 patients meeting with the NINCDS-ARDRA criteria of AD were divided into 2 equal groups. Patients in the capsule group received 4 capsules of Hup (each contains 50 micrograms) and 4 tablets of placebo (lactose and starch inside); while the tablet group received 4 tablets of Hup (each contains 50 micrograms) and 4 capsules of placebo, p.o., twice a day for 60 d. All the patients were evaluated with a lot of related ranting scales, and physiological and laboratory examination. RESULTS: There were significant differences (P < 0.01) on all the psychological evaluations between 'before' and 'after' the 60-d trial of 2 groups, but there was no significant difference between 2 groups by group t test (P > 0.05). The changes of oxygen free radicals in 2 groups showed marked improvement. No severe side effect besides moderate to mild nausea was found in both groups. CONCLUSION: There is equal efficacy and safety between Hup in capsule and tablet for treating patients with AD, and Hup can reduce the pathological changes of the oxygen free radicals in the plasma and erythrocytes of patients with AD.