[Pharmacokinetics of 11 active components of Psoraleae Fructus in normal and diabetic rats].

A total of 11 active ingredients including psoralen, isopsoralen, bakuchiol, bavachalcone, bavachinin, corylin, coryfolin, isobavachalcone, neobavaisoflavone, bakuchalcone, and corylifol A from Psoraleae Fructus in the plasma samples of diabetic and normal rats were simultaneously determined by UHPLC-MS/MS. The pharmacokinetic parameters were calculated to elucidate the pharmacokinetic profiles of coumarins, flavonoids, and monoterpene phenols in normal and diabetic rats. The rat model of type 2 diabetes mellitus(T2DM) was induced by a high-sugar and high-fat diet combined with injection of 1% streptozotocin every two days. The plasma samples were collected at different time points after the rats were administrated with Psoraleae Fructus. The proteins in the plasma samples were precipitated by ethyl acetate, and the plasma concentrations of the 11 components of Psoraleae Fructus were determined by UHPLC-MS/MS. The pharmacokinetic parameters were calculated by DAS 3.0. The results showed that the pharmacokinetic beha-viors of 8 components including psoralen, isopsoralen, bakuchiol, and bavachinin from Psoraleae Fructus in both female and male mo-del rats were significantly different from those in normal rats. Among them, the coumarins including psoralen, isopsoralen, and corylin showed lowered levels in the blood of both female and male model rats. The flavonoids(bavachinin, corylifol A, and bakuchalcone) and the monoterpene phenol bakuchiol showed decreased levels in the female model rats but elevated levels in the male model rats. It is suggested that the dosage of Psoraleae Fructus should be reasonably adjusted for the patients of different genders at the time of clinical administration.

Dihydromyricetin Alleviates H9C2 Cell Apoptosis and Autophagy by Regulating CircHIPK3 Expression and PI3K/AKT/mTOR Pathway.

OBJECTIVE: To investigate the effect and potential mechanism of dihydromyricetin (Dmy) on H9C2 cell proliferation, apoptosis, and autophagy. METHODS: H9C2 cells were randomly divided into 7 groups, namely control, model, EV (empty pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector), IV (circHIPK3 interference), Dmy (50 µ mol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 II/I (LC3II/I), phospho-phosphoinositide 3-kinase (p-PI3K), protein kinase B (p-AKT), and phospho-mammalian target of rapamycin (p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells. RESULTS: Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly (P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3 II/I significantly increased (all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3 II/I decreased significantly (all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced (P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed (P<0.05). CONCLUSION: Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.

Preliminary verification of the anti-hypoxia mechanism of Gentiana straminea maxim based on UPLC-triple TOF MS/MS and network pharmacology.

BACKGROUND: Anoxia is characterized by changes in the morphology, metabolism, and function of tissues and organs due to insufficient oxygen supply or oxygen dysfunction. Gentiana straminea Maxim (G.s Maxim) is a traditional Tibetan medicine. Our previous work found that G.s Maxim mediates resistance to hypoxia, and we found that the ethyl acetate extract had the best effect. Nevertheless, the primary anti-hypoxia components and mechanisms of action remain unclear. METHODS: Compounds from the ethyl acetate extraction of G.s Maxim were identified using UPLC-Triple TOF MS/MS. Then Traditional Chinese Medicine Systematic Pharmacology Database was used to filtrate them. Network pharmacology was used to forecast the mechanisms of these compounds. Male specific pathogen-free Sprague Dawley rats were randomly divided into six groups: (1) Control; (2) Model; (3) 228 mg/kg body weight Rhodiola capsules; (4) 6.66 g/kg body weight the G.s Maxim's ethyl acetate extraction; (5) 3.33 g/kg body weight the G.s Maxim's ethyl acetate extraction; (6) 1.67 g/kg body weight the G.s Maxim's ethyl acetate extraction. After administering intragastric ally for 15 consecutive days, an anoxia model was established using a hypobaric oxygen chamber (7000 m, 24 h). Then Histology, enzyme-linked immunosorbent assays, and western blots were performed to determine these compounds' anti-hypoxic effects and mechanisms. Finally, we performed a molecular docking test to test these compounds using Auto Dock. RESULTS: Eight drug-like compounds in G.s Maxim were confirmed using UPLC-Triple TOF MS/MS and Lipinski's rule. The tumor necrosis factor (TNF) signaling pathway, the hypoxia-inducible factor 1 (HIF-1) signaling pathway, and the nuclear factor kappa-B (NF-κB) signaling pathway was signaling pathways that G.s Maxim mediated anti-anoxia effects. The critical targets were TNF, Jun proto-oncogene (JUN), tumor protein p53 (TP53), and threonine kinase 1 (AKT1). Animal experiments showed that the ethyl acetate extraction of G.s Maxim ameliorated the hypoxia-induced damage of hippocampal nerve cells in the CA1 region and reversed elevated serum expression of TNF-α, IL-6, and NF-κ B in hypoxic rats. The compound also reduced the expression of HIF-1α and p65 and increased the Bcl-2/Bax ratio in brain tissue. These findings suggest that G.s Maxim significantly protects against brain tissue damage in hypoxic rats by suppressing hypoxia-induced apoptosis and inflammation. Ccorosolic acid, oleanolic acid, and ursolic acid had a strong affinity with core targets. CONCLUSIONS: The ethyl acetate extraction of G.s Maxim mediates anti-hypoxic effects, possibly related to inhibiting apoptosis and inflammatory responses through the HIF-1/NF-κB pathway. The primary active components might be corosolic, oleanolic, and ursolic acids.

A pair of novel phenylethanol glycosides from Cistanche tubulosa (Schenk) Wight.

A pair of differential epimers with opposite C-7 configurations, crenatosides A and B (1 and 2), and 10 known phenylethanoid glycosides (PhGs) (3-12) were obtained from the succulent stem of Cistanche tubulosa. The structures were elucidated based on extensive spectral data (UV, IR, 1D and 2D NMR, HR-ESIMS), which are first reported natural products with unique glycoside structures. After acid hydrolysis, the configuration of the sugar was determined by comparing it with the normative sugar by HPLC. The absolute configurations of both compounds were determined by ECD spectrum analysis. All the obtained compounds were examined for their inhibitory effect on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in mouse microglial cells (BV-2 cells), and compounds 1 and 2 showed potent inhibition on NO production with IC50 values of 5.62 µM and 6.30 µM, respectively.

Study on pharmacokinetics of eight active compounds from Bufei-Huoxue Capsule based on UHPLC-MS/MS.

Bufei-Huoxue Capsule (BFHX) was applied to treat chronic obstructive pulmonary disease (COPD) in China. It is composed of Astragali Radix, Paeoniae Radix Rubra, and Psoralea Fructus. A sensitive and reliable ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) method was developed and validated to quantify the eight main bioactive compounds (psoralen, isopsoralen, neobabaisoflavone, corylin, bavachin, astragaloside IV, ononin and formononetin) in rat plasma after oral administration of BFHX. Osthol was used as an internal standard (IS). Plasma samples were pretreated with methanol to precipitate protein. Chromatographic separation was accomplished using Hypersil GOLDTM C18 column (2.1 mm × 100 mm, 1.9 µm) with a gradient elution profile and a mobile phase consisting of (A) 0.1% formic acid in water and (B) acetonitrile and the flow rate was set at 0.2 mL/min. Multiple reaction monitoring (MRM) mode was applied to perform mass spectrometric analyses. All calibration curves were linear (r > 0.9908) in tested ranges. The intra- and inter-day accuracy and precisions of eight compounds at three different concentration levels were within the acceptable limits. The extraction recovery was within the range of 76.4 âˆ¼ 105.2% and the matrix effects were within the range of 88.3 âˆ¼ 115.0% (RSD ≤ 15.6%). The dilution effects were within the range of 90.2 âˆ¼ 114.9%. These 8 compounds were stable under the tested conditions. So the developed method was valid to evaluate the pharmacokinetic study of eight bioactive compounds after oral administration of BFHX.

Effectiveness of Melodic Intonation Therapy in Chinese Mandarin on Non-fluent Aphasia in Patients After Stroke: A Randomized Control Trial.

Melodic intonation therapy (MIT) positively impacts the speech function of patients suffering from aphasia and strokes. Fixed-pitch melodies and phrases formulated in MIT provide the key to the target language to open the language pathway. This randomized controlled trial compared the effects of music therapy-based MIT and speech therapy on patients with non-fluent aphasia. The former is more effective in the recovery of language function in patients with aphasia. Forty-two participants were enrolled in the study, and 40 patients were registered. The participants were randomly assigned to two groups: the intervention group (n = 20; 16 males, 4 females; 52.90 ± 9.08 years), which received MIT, and the control group (n = 20; 15 males, 5 females; 54.05 ± 10.81 years), which received speech therapy. The intervention group received MIT treatment for 30 min/day, five times a week for 8 weeks, and the control group received identical sessions of speech therapy for 30 min/day, five times a week for 8 weeks. Each participant of the group was assessed by a Boston Diagnostic Aphasia Examination (BDAE) at the baseline (t1, before the start of the experiment), and after 8 weeks (t2, the experiment was finished). The Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD) were also measured on the time points. The best medical care of the two groups is the same. Two-way ANOVA analysis of variance was used only for data detection. In the spontaneous speech (information), the listening comprehension (right or wrong, word recognition, and sequential order) and repetitions of the intervention group were significantly higher than the control group in terms of the cumulative effect of time and the difference between groups after 8 weeks. The intervention group has a significant time effect in fluency, but the results after 8 weeks were not significantly different from those in the control group. In terms of naming, the intervention group was much better than the control group in spontaneous naming. Regarding object naming, reaction naming, and sentence completing, the intervention group showed a strong time accumulation effect. Still, the results after 8 weeks were not significantly different from those in the control group. These results indicate that, compared with speech therapy, MIT based on music therapy is a more effective musical activity and is effective and valuable for the recovery of speech function in patients with non-fluent aphasia. As a more professional non-traumatic treatment method, MIT conducted by qualified music therapists requires deeper cooperation between doctors and music therapists to improve rehabilitating patients with aphasia. The Ethics Committee of the China Rehabilitation Research Center approved this study (Approval No. 2020-013-1 on April 1, 2020) and was registered with the Chinese Clinical Trial Registry (Registration number: Clinical Trials ChiCTR2000037871) on September 3, 2020.

[Acupuncture of "Zhibian (BL 54) through Shuidao (ST 28)" for female stress urinary incontinence: a randomized controlled trial].

OBJECTIVE: To observe effect of "Zhibian (BL 54) through Shuidao (ST 28)" acupuncture for quality of life of female patients with stress urinary incontinence. METHODS: A total of 90 female patients were randomly divided into an observation group and a control group, 45 cases in each group. The patients in the observation group were treated with "Zhibian (BL 54) through Shuidao (ST 28)" acupuncture method, and the patients in the control group were treated with non-permeable sham acupuncture at Zhibian (BL 54). The needles were retained for 30 min in both groups, once a day, and the treatment was totally given 10 times. The score of urinary incontinence quality of life questionnaire (I-QOL) was recorded before and after treatment and during the follow-up 1 month after treatment in the two groups, and the 1 h urine pad test and the 72 h urination diary card were used to evaluate the 1 h urine leakage and the 24 h urine leakage frequency of the two groups. RESULTS: After treatment and during follow-up, the I-QOL scores in the observation group were higher than those before treatment (P<0.05), and were higher than those in the control group (P<0.05). After treatment and during follow-up, the 1 h urine leakage and the 24 h urine leakage frequency in the observation group were lower than those before treatment (P<0.05), and less than those in the control group (P<0.05). CONCLUSION: Acupuncture of "Zhibian (BL 54) through Shuidao (ST 28)" can improve the quality of life of female patients with stress urinary incontinence, and improve the volume and frequency of urine leakage.

Proteome and Transcriptome Analysis of the Antioxidant Mechanism in Chicken Regulated by Eucalyptus Leaf Polyphenols Extract.

Eucalyptus leaf polyphenols extract (EPE) has been proved to have various bioactivities, but few reports focus on its antioxidant mechanism in vivo. The purpose of this study was to elucidate the effect and mechanism of EPE dietary supplements on antioxidant capacity in chicken. A total of 216 chickens were randomly selected for a 40-day experiment. Four treatment groups received diets including the control diet only, the control diet + low EPE (0.6 g/kg), the control diet + moderate EPE (0.9 g/kg), and the control diet + high EPE (1.2 g/kg). Compared with control group, the glutathione peroxidase (GSH-Px) activity and glutathione (GSH) content in the breast muscle of the moderate EPE treatment group was significantly higher (p < 0.05), while the malonaldehyde (MDA) content in the moderate EPE group was reduced (p < 0.05). Moreover, proteomic and transcriptomic analyses of the breast muscle revealed that glutathione metabolism and the peroxisome were the two crucial metabolic pathways responsible for increased antioxidant capacity of the muscle. Accordingly, nine candidate genes and two candidate proteins were identified related to improved antioxidant status induced by EPE supplements. This research provides new insights into the molecular mechanism of antioxidant capacity in chickens treated with EPE dietary supplements.

RNA-seq-based quanitative transcriptome analysis of meat color and taste from chickens administered by eucalyptus leaf polyphenols extract.

To evaluate how eucalyptus leaf polyphenol extract (EPE) affects chicken meat color and taste, we added different levels of EPE (0%, 0.06%, 0.09%, and 0.12%) to chicken feed. The redness (a* value) and the myoglobin content of breast muscle in EPE group were remarkably higher. Furthermore, the guanosine monophosphate, histidine, and glycine muscle contents were also enhanced. Transcriptome analysis showed that 10 candidate genes related to meat quality were affected by EPE treatment. The identified genes, with functions critical to chicken meat color and taste, will help to determine the molecular mechanisms of EPE.

Chai-Qin-Cheng-Qi Decoction and Carbachol Improve Intestinal Motility by Regulating Protein Kinase C-Mediated Ca2+ Release in Colonic Smooth Muscle Cells in Rats with Acute Necrotising Pancreatitis.

Chai-Qin-Cheng-Qi decoction (CQCQD) improves intestinal motility in acute pancreatitis (AP), but the mechanism(s) require elucidation. We investigated the effects of CQCQD and carbachol, a prokinetic agent, on colonic smooth muscle cells (SMCs) in L-arginine-induced necrotising AP model in rats. In treatment groups, intragastric CQCQD (20 g/kg, 2 hourly × 3 doses) or intraperitoneal carbachol (60 µg/kg) was given 24 hours after induction of AP. Both CQCQD and carbachol decreased the severity of pancreatic and colonic histopathology (all P < 0.05). Both CQCQD and carbachol reduced serum intestinal fatty acid binding protein, vasoactive intestinal peptide, and substance P and increased motility levels. CQCQD upregulated SMC phospholipase C-beta 1 (PLC-ß1) mRNA and PLC protein (both P < 0.05), while both treatments upregulated protein kinase C-alpha (PKC-α) mRNA and PKC protein and downregulated adenylate cyclase (AC) mRNA and protein compared with no treatment (all P < 0.05). Neither treatment significantly altered L-arginine-induced PKC-ß1 and PKC-ε mRNA reduction. Both treatments significantly increased fluorescence intensity of SMC intracellular calcium concentration [Ca2+]i (3563.5 and 3046.9 versus 1086.9, both P < 0.01). These data suggest CQCQD and carbachol improve intestinal motility in AP by increasing [Ca2+]i in colonic SMCs via upregulating PLC, PKC and downregulating AC.

Scorpion venoms in gastric cancer.

Venom secretions from snakes, scorpions, spiders and bees, have been widely applied in traditional medicine and current biopharmaceutical research. Possession of anticancer potential is another novel discovery for animal venoms and toxins. An increasing number of studies have shown the anticancer effects of venoms and toxins of snakes, and scorpions in vitro and in vivo, which were achieved mainly through the inhibition of cancer growth, arrest of cell cycle, induction of apoptosis and suppression of cancer metastasis. However, more evidence is needed to support this concept and the mechanisms of anticancer actions are not clearly understood. The present review is focused on the recant updates on anticancer venom research.

[Effects of qishenyiqi gutta pills on calcium/calmodulin dependent protein kinase II in rats with renal hypertension].

OBJECTIVE: To explore the effects of qishenyiqi gutta pills on myocardial hypertrophy of left ventricle and calcium/calmodulin dependent protein kinase II (CAMK II) in rats with renal hypertension and elucidate its intervention mechanism for myocardial hypertrophy. METHODS: A total of 50 Wistar rats were randomly divided into 5 groups of sham-operation, control, high-dose qishenyiqi gutta pills, low-dose qishenyiqi gutta pills and valsartan (n = 10 each). The rat model of myocardial hypertrophy with renal hypertension was established by the 2-kidney 1-clip (2K1C) method. The experimental animals were divided into control, high-dose, low-dose and valsartan groups. At Week 5 postoperation, valsartan group received an oral dose of valsartan (30 mg×kg(-1)×d(-1)), high-dose and low-dose groups took qishenyiqi gutta pills (250 and 125 mg×kg(-1)×d(-1)) while sham-operation and control groups had the same dose of normal saline solution. Tail arterial pressure was detected weekly and continued for 8 weeks. At the end of Week 12, the animals were sacrificed to harvest myocardial tissue of left ventricle for detecting left ventricular mass index (LVMI). The collagen volume fraction (CVF) of myocardium was examined by Van Gieson staining, the activities of superoxide dismutase (SOD) and reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA) and the expression of CAMK II was detected by immunohistochemistry and Western blot. RESULTS: (1) Blood pressures were significantly higher in high-dose, low-dose and control groups than those in sham-operation and valsartan groups ((167.66 ± 11.48), (166.72 ± 13.51), (174.34 ± 14.52) vs (119.57 ± 6.30), (131.80 ± 12.49) mm Hg, P < 0.01). The changes of blood pressure had no significant difference between high-dose and low-dose groups. (2) LVMI and CVF increased significantly in high-dose, low-dose and valsartan groups versus sham-operation group (LVMI: (1.98 ± 0.16), (2.09 ± 0.14), (1.97 ± 0.17) vs (1.74 ± 0.17) g/kg; CVF: 0.94% ± 0.22%, 2.53% ± 0.61%, 0.81% ± 0.20% vs 0.45% ± 0.13%) (P < 0.01, P < 0.05), but decreased significantly versus control group (LVMI: (1.98 ± 0.16), (2.09 ± 0.14), (1.97 ± 0.17) vs (2.28 ± 0.28) g/kg; CVF: 0.94% ± 0.22%, 2.53% ± 0.61%, 0.81% ± 0.20% vs 4.73% ± 1.04%) (P < 0.01, P < 0.05). (3) The expression of CAMK II was significantly higher in high-dose, low-dose, valsartan and control groups than that in sham-operation group (65.9%, 95.3%, 84.8%, 160.1% vs 67.7%). And it was significantly lower in high-dose, low-dose and valsartan groups than that in control group (65.9%, 95.3%, 84.8% vs 160.1%). There was no statistical difference among high-dose, low-dose and valsartan groups. CONCLUSIONS: Qishenyiqi gutta pills may retard myocardial hypertrophy of left ventricle in rats with renal hypertension. And the mechanism is probably be correlated with its antioxidant activity and inhibited expression of myocardial CAMK II.

The role of ß-catenin signaling pathway on proliferation of rats neural stem cells after hyperbaric oxygen therapy in vitro.

ß-catenin, a protein that functions in both cell adhesion and Wnt signaling, plays a key role in mammalian neural development. To investigate the role of ß-catenin in hyperbaric oxygen therapy (HBO)-induced neurogenesis after hypoxic ischemic brain damage (HIBD), we transfected ß-catenin siRNA and negative control siRNA into neural stem cells (NSCs) after HIBD. We found that HBO promoted NSCs differentiate into neurons or oligodendrocytes, and inhibited NSCs differentiate into astrocytes; HIBD brain tissue extract conditioned cultures promoted NSCs differentiate into neurons; ß-Catenin siRNA decreased the NSE-positive neurons and increased GFAP-positive astrocytes in the NSCs in vitro. Furthermore, the expression of Ngn1 protein and mRNA in NSCs was increased when HBO promoted NSCs differentiate into neurons after HIBD, and the expression of BMP-4 protein and mRNA was decreased when HBO depressed NSCs differentiate into astrocytes after HIBD. These results showed that ß-catenin-mediated transcriptional activation functions in the decision of NSCs to proliferate neurogenesis during HBO-induced after HIBD, and suggested that HBO therapy promotes the proliferation of neural stem cells in vitro, an effect that may be correlated with ß-catenin protein and HBO therapy could promote neurogenesis by ß-catenin-induced activated Ngn1 gene and repress astrocytogenesis by ß-catenin-induced down-regulated BMP-4 gene.

[Studies on the chemical constituents from Xinjiang Punica granatum].

OBJECTIVE: To study the chemical constituents from Punica granatum. METHODS: The chemical constituents were isolated by column chromatography on silica gel and MCI-gel CHP 20P, Sephadex LH-20. Their structures were elucidated on the basis of spectral analysis and physical constants. RESULTS: Ten compounds were isolated and elucidated as: gallic acid (1), methyl gallate (2), ellagic acid (3), (+) catechin (4), isoquerecitrin (5), D-mannitol (6), ursolic acid (7), Oleanolic acid (8), beta-Sitosterol (9) and Daucosterol (10). CONCLUSION: Compounds 1-10 are isolated from this plant for the first time.

[Immunological mechanism of exfoliative tongue fur in children with asthma].

OBJECTIVE: To explore the immunological mechanism of exfoliative tongue fur in children with asthma. METHODS: Thirty-nine children with asthma, twenty-eight children with repetitive respiratory tract infection (non-asthma) and eleven healthy children were divided into five groups, which were asthma with exfoliative fur or with non-exfoliative fur groups, non-asthma with exfoliative fur or with non-exfoliative fur groups and normal control group. The concentrations of keratin 13 and bcl-2 in cells exfoliated from tongue fur were detected by immunohistochemical method. The expression levels of blood cell chemokine receptor-3 (CCR-3) and CD4(+) were examined by flow cytometry, and the levels of serum cortisol and IgE were detected by radioimmunoassay. RESULTS: The levels of blood CD4(+) and CCR-3 of children with asthma and exfoliative fur were higher than those in the asthma with non-exfoliative fur group and the normal control group (P<0.05). The serum level of cortisol in the groups of asthma with exfoliative fur and non-asthma with exfoliative fur were lower than that in the other groups (P<0.05). The serum levels of IgE in asthma with exfoliative fur or with non-exfoliative fur groups were higher than that in the other groups (P<0.05). Concentrations of keratin 13 in the cells exfoliated from tongue fur in the groups of asthma or non-asthma with exfoliative fur were lower than that of the other groups (P<0.05). There was no significant difference of expression level of bcl-2 in the cells exfoliated from tongue fur among these five groups. CONCLUSION: There is a reasonably close relationship between the formation of exfoliative tongue fur and the immune system such as low level of serum cortisol and high levels of blood CD4(+) and CCR-3, which may all promote the formation of exfoliative fur. The disability of keratinization and apoptosis of epithelial cells of tongue may also be one cause for its formation.

[Effect of Bushen recipe on T-cell subsets and their function in patients with chronic hepatitis B].

OBJECTIVE: To observe the regulatory effect of Bushen recipe (BSR) on immune function in patients with chronic hepatitis B. METHODS: Thirty patients with HBeAg+ and abnormal alanine transaminase (ALT) level were treated with BSR for 6 months. The following parameters before treatment, 3 months and 6 months after treatment were tested: (1) HBV DNA by fluorescence quanitiative PCR; (2) Levels of interleukin-10 (IL-10) and interferon-gamma in supernatant of cultured peripheral blood mononuclear cell (PBMC) after had been cultured with HBeAg, HBcAg and phytohemagglutinin (PHA) for 48 hrs; (3) Blood levels of T-cells, including CD4+ , CD8+ , CD8+ CD28+ , CD8+ CD28- and CD28+ , and their ratio, as well as the percentage of CD8+ CD28+ in the cell culture after being co-cultured with PHA for 48 hrs. RESULTS: In patients effectively treated by BSR, after 3 months treatment, level of IFN-gamma in PBMC culture significantly increased, CD8+ CD28+ , CD28+ T-cells significantly enhanced and IL-10 and CD8 + CD28+ T-cells significantly decreased (P < 0.05, P < 0.01). CONCLUSION: BSR could significantly strengthen the expression of Th1 type cytokine, decrease the expression of Th2 type cytokine and promote the expression of CD8+ cytotoxic T-lymphocyte, which may be one of mechanisms in suppressing HBV DNA replication.

[Relationship between CD8+ T-cell CD28 expression and TCM differentiation type in patients with chronic aplastic anemia].

OBJECTIVE: To explore the relationship between CD8+ T-cell CD28 molecular expression in peripheral blood and TCM type in patients with chronic aplastic anemia (CAA). METHODS: Using flow cytometry to detect the CD28 expression in 45 in-patients or out-patients and 24 healthy subjects for control. And the relation with TCM type was analyzed from the immunological aspect. RESULTS: (1) The levels of CD8, CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients than those in the healthy subjects (P < 0.05 or P < 0.01). (2) The levels of CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients of Shen-Yin deficiency type than those in the CAA patients of Shen-Yang deficiency type (P < 0.05 or P < 0.01). CONCLUSION: (1) The abnormal high expression of peripheral blood co-stimulatory molecules CD28 suggested CD28 disorder may play an important role in immuno-pathogenesis of CAA. (2) The levels of peripheral CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- can be taken as an objective indexes for TCM typing of CAA, which was disordered more severe in patients of Shen-Yin deficiency type than in those of Shen-Yang deficiency type.

[Role of cyclin kinase inhibitor p27 in inhibition of emodin on mesangial cell proliferation induced by tumor necrosis factor-alpha].

OBJECTIVE: To investigate the role of p27 in the inhibition of emodin on the mesangial cell (MC) proliferation induced by tumor necrosis factor-alpha(TNF-alpha). METHODS: p27 protein of MC was detected with western blotting analysis. The degree of MC proliferation was estimated through [(3)H] thymidine ([(3)H] TdR) incorporation. Different dosage of emodin (50 mg/L,100 mg/L) was added into MC stimulated by TNF-alpha. RESULTS: TNF-alpha (200 kU/L) decreased p27 level of MC cultured in serum-free DMEM for 24 hours and increased[(3)H] TdR incorporation. Emodin increased p27 level of MC stimulated by TNF-alpha and decreased [(3)H] TdR incorporation. The more the emodin was added, the greater the above-mentioned effect of emodin. CONCLUSION: The increment of p27 level maybe play an important role in the inhibition of emodin on MC proliferation induced by TNF-alpha.