Object To apply microwave technique to the extraction of free anthraquinones from Rheum emodi Wall. and the methods of microwave extraction and traditional extraction were comparied. Methods The orthogonal test design was used to arrange sequent experiments to test the effect of the three main factors including microwave power, particle size of herb and extraction time on the extraction efficiency of free anthraquinones and to establish optimal extraction process. A parallel test was carried out using the three different methods including the optimal extraction process, the traditional decocting method and the alcohol refluxing method. Results Particle size of herb and microwave power are the significant factors with P

AIM: To optimize and establish the extraction techn ology of water-soluble polysaccharides from Ulva pertusa. TLC was employed to determine the mainly neutral sugars in the Ulvan. METHODS: The orthogonal test was employed to test the effects of th e four factors including volume of water, temperature of extraction, time of ext raction and pH of water on the yield and total carbohydrate contents of water-s oluble polysaccharides from Ulva pertusa and optimize the extraction technol ogy. Five groups of parallel tests were carried out by the optimum parameters es tablished and the sugar constituents of products were analyzed by TLC. RESULTS: Temperature of extraction and time of extraction have sign ificant effects on the extraction yield of water-soluble polysaccharides from Ulva pertu sa, but, for total carbohydrate content, only temperature of extraction was a significant factor. The mainly neutral sugars were composed of rhamnose, xylose and glucose. CONCLUSION: The optimum extraction technology was A 1B 3C 3D 3 when the yield and total carbohydrate content were considered as preferential pa rameters.

Objective To analyze the property of natural polysaccharides(P) from Porphyra haitanensis T. J. Chang et B. F. Zheng and its degraded polysaccharides(P3),and to investigate their effects on proliferation of lymphocytes. Methods P3 and P were identified by IR spectra. Their effects on immunological function were studied by cell culture technique. Results P3 and P displayed significant effects on promoting the proliferation of T-lymphocytes,and had inhibitory effects on the proliferation of B-lymphocytes in a dose-dependent manner. The effects of P3 were more powerful than P. Conclusion The degraded and natural polysaccharides from Porphyra haitanensis(P3 and P) can regulate the proliferation of lymphocytes,which may be related to their molecular weights.

BACKGROUND:It is unclear whether serial cel passage in vitro influences the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s.
OBJECTIVE:To investigate the effect of cel passage on the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s.
METHODS:Rat bone marrow mesenchymal stem cel s were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cel s at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cel biological characterization was observed and differenitaiton ability into neural stem cel s was observed by detecting Nestin expression in cel s using flow cytometry. Then, the cel s were further induced to differentiate and cel multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression.
RESULTS AND CONCLUSION:Under induction, bone marrow mesenchymal stem cel s at different passages were al differentiated into Nestin-positive neural stem cel s. However, there was a significant difference in differentiation proportion of cel s at different passages (P<0.05). Strongest differentiation ability was found in the passage 6 cel s, with the Nestin expression up to (93.7±2.3)%at 7 days of induction and (96.2±1.8)%at 14 days of induction. The proportion of differentiated cel s at passages 6 and 9 was signfi cantly higher than that at passages 3 and 12. Moreover, adherent cel s were positive for nerve enolase and glial acidic protein. Al these findings indicate that the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s is correlated with cel passage. Cel s at lower or higher passages are both detrimental to cel differentiation.

Objective To investigate the effects of collateral pathways on prognosis of cerebral infarct patients with prior transient ischemic attack (TIA) and its role in neuroprotective mechanism of TIA. Methods A total of 164 cases of the first ever cerebral infarct patients were consecutively allocated into three groups: A , B and C group according to their age.Three groups were divided into two subgroups respectively based on the absence or presence of prior ipsilateral TIA: A1, A2, B1, B2, C1, C2 group. Neurological dysfunctional scores at admission and 1 month after treatment, barthel index, collateral pathways status, and cerebral infarction volume were evaluated respectively. The relationship between development of collateral pathways and prognosis was assessed at the same time. Results The neurological dysfunctional scores and cerebral infarction volume of patients in A1 group and B1 group were significantly lower than those of A2 group and B2 group (P

BACKGROUND: There are numerous inducers used in inducing bone marrow mesenchymal stem cells (BMMSCs) differentiate into neuron-like cells, however, due to poisonous, most chemical inducers can not be used in human.OBJECTIVE: To investigate the effect of ligustrazine on differentiation of rat BMMSCs into neuron-like cells in vitro, and to search for the optimal inductive concentration.METHODS: After SD rats were anesthetized, bone marrow was obtained from the femoral and tibial bones, centrifuged, and the supernatant was discarded. The extracted cells were cultured in L-DMEM supplemented with 15% fetal bovine serum. The expression of CD44 and CD45 of the 5~(th) passage of BMMSCs were identified by immunocytochemical technique. Serum-free L-DMEM medium contains 1.00, 1.25 and 1.50 g/L ligustrazine concentrations were used to induce the 5~(th) passage of BMMSCs in vitro. Morphology changes of BMMSCs were observed under an inverted phase microscope. Expression of nestin, neuron-specific enolase and glial fibrillary acidic protein were identified by immunocytochemical technique, and the expression ratio of neuron-like cells' surface antigens induced by different concentrations of ligustrazine were compared.RESULTS AND CONCLUSION: ①Most primarily cultured BMMSCs adhered to the wall at 3 days after culture, which proliferated faster after passaged, and the 5~(th) passage of cells were mostly purified into BMMSCs, spread radially or vortex-likely. ②The 5~(th) passage of BMMSCs was positive expressed (98.02±0.81)% CD44, but negative for CD45. ③Neuron-like cells with prominence and bifurcation could be seen after induction. The immunocytochemical method showed that nestin and neuron-specific enolase in most induced cells were positive expressed, especially received a highest ration of neuron-specific enolase expressing in the induced group with 1.25 g/L concentration of ligustrazine. It revealed that ligustrazine can induce BMMSCs differentiated into neuron-like cells, and 1.25 g/L is the optimal inductive concentration.

Abstract: Objective To research the relationship between promoter polymorphism-438 A/G of thrombin-activatable fibrinolysis inhibitorgene (TAFI-438 A/G) and atherosclerotic cerebral infarction (ACI) in Chinese Han population. Methods TAFI-438A/G genotypes andtheir allele frequencies were identified with the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in225 ACI patients and 184 healthy controls. Results There was no significant difference of TAFI-438A/G polymorphism between ACI groupand control group. Stratified by gender, in males, the incidence of A allele was 28.6% in ACI group, and 20.6% in control (P=0.039); of theAA genotype was 9.0% in ACI group, and 1.9% in control (P=0.019). But no significant difference was found in females. ConclusionTAFI-438A/G polymorphism is associated with the risk of ACI in males, that AA genotype may increase the risk of ACI.

ObjectiveTo investigate the relationship between Parkinson disease (PD) and intron rs12138592 A/G polymorphism of ubiquitin specific proteases (USP24) gene in Han population of the Western Guangdong province in China. Methods81 PD cases and 100 ethnically matched controls were investigated USP24 gene rs12138592 A/G polymorphism with polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). ResultsThe incidence of GG genotype was 77.8% in the cases and 62.0% in the controls (χ2=5.213,P=0.022), and the G allele was 88.3% in the cases, 79.5% in the controls (χ2=4.980，P=0.026). ConclusionThe G allele and GG genotype of USP24 gene rs12138592 A/G polymorphism can increase the risk of suffering from PD.

Objective:To investigate the effect and mechanism of Sufentanil (Suf) on oxidative stress injury in osteoporosis (OP) rats.Methods:Rats were randomly separated into normal group, OP group, low concentration Suf group, high concentration Suf group, alendronate sodium group, and high concentration Suf+AMPK inhibitor (Dorsomorphin) group, with 12 rats in each group. The normal group of rats only removed a small amount of adipose tissue around the ovaries. Except for the normal group, rats in other groups constructed OP models by removing both ovaries. After successful modeling, they were administered once a day for 12 weeks. Micro CT scanners were applied to measure bone density and trabecular structure parameters in rats, including trabecular quantity, trabecular thickness, and trabecular separation. ELISA was applied to detect levels of osteocalcin, alkaline phosphatase (ALP), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-B ligand (RANKL) in rat serum. HE staining was applied to detect pathological changes in femoral tissue. Reagent kit was applied to detect levels of malondialdehyde (MDA), reactive oxygen species (ROS), and superoxide dismutase (SOD) in femoral tissue. Western blot analysis of phosphorylated adenylate activated protein kinase (p-AMPK), peroxisome proliferator-activated receptor-γ-coactivator 1α (PGC-1α), silence-information regulatory factor2-associated enzyme 1 (SIRT1) proteins in femur tissue.Results:Compared with the normal group, the bone trabecular structure of rats in the OP group was thin and scattered, with fewer bone trabeculae and widened trabecular gaps, mainly in the form of rods, the bone density, number of trabeculae, thickness of trabeculae, levels of ALP, osteocalcin, OPG in serum, the level of SOD, and p-AMPK, PGC-1α, and SIRT1 proteins in femoral tissue decreased ( q=38.00, 67.81, 47.91, 48.32, 28.42, 21.23, 52.22, 30.86, 24.11, 24.55, P all<0.001), the bone trabecular separation, level of RANKL in serum, and the levels of MDA and ROS in femoral tissue increased ( q=42.00, 37.97, 50.56, 72.01, P all<0.001). Compared with the OP group, the number of bone trabeculae in rats in the low concentration Suf group, high concentration Suf group, and alendronate sodium group increased, the gap decreased, and the bone trabecular structure gradually recovered to a plate-like shape, the bone density, number of trabeculae, thickness of trabeculae, levels of ALP, osteocalcin, OPG in serum, the level of SOD, and p-AMPK, PGC-1α, and SIRT1 proteins in femoral tissue increased ( F=214.40, 889.60, 396.10, 480.30, 168.60, 38.09, 367.80, 187.70, 91.17, 101.00, P all<0.001), the bone trabecular separation, level of RANKL in serum, and the levels of MDA and ROS in femoral tissue decreased ( F=174.10, 173.40, 298.90, 607.10, P all<0.001). Dorsomorphin reversed the improvement effect of high concentration of Suf on oxidative stress injury in OP rats. Conclusion:The inhibitory effect of Suf on oxidative stress injury in OP rats may be related to the activation of the AMPK/SIRT1/PGC-1α signaling pathway.

Objective:To observe the effects of self-made Qingyuan Shenghua decoction on coagulation dysfunction in patients with sepsis, and to explore its possible mechanism.Methods:Eighty patients with sepsis and coagulation dysfunction admitted to the department of critical care medicine of Chengdu First People's Hospital from March 2018 to April 2020 were enrolled. The patients were divided into control group and observation group according to random number table method, with 40 cases in each group. Patients in both groups received basic treatment for sepsis. On this basis, the observation group was administrated with self-made Qingyuan Shenghua decoction, one dose a day, 100 mL in the morning and 100 mL in the evening; the control group was given the same amount of normal saline. Both groups were treated for 7 days. Prothrombin time (PT), activated partial thromboplastin time (APTT), international normalized ratio (INR), fibrinogen (Fib), D-dimer, platelet count (PLT), white blood cell count (WBC), C-reactive protein (CRP), and procalcitonin (PCT) were measured before and after treatment, and acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and sequential organ failure assessment (SOFA) were calculated. The length of intensive care unit (ICU) stay, the incidence of multiple organ dysfunction syndrome (MODS) and 28-day mortality was recorded.Results:The indexes of coagulation function and inflammation in the two groups were significantly improved after treatment, the improvement of various indexes in the observation group were better than those in the control group [PT (s): 16.01±1.08 vs. 19.21±1.38, APTT (s): 55.33±15.29 vs. 79.41±12.69, INR: 1.30±0.21 vs. 1.65±0.22, Fib (g/L): 2.87±0.89 vs. 5.44±1.13, D-dimer (mg/L): 2.56±1.67 vs. 6.41±2.42, PLT (×10 9/L): 125.79±18.51 vs. 95.46±18.50, WBC (×10 9/L): 7.50±0.78 vs. 12.75±4.09, CRP (mg/L): 21.27±9.32 vs. 65.44±13.40, PCT (μg/L): 1.15±0.58 vs. 6.31±1.29], and the differences were statistically significant (all P < 0.05). After treatment, APACHEⅡ and SOFA scores in the two groups decreased significantly compared with those before treatment, and the decrease in the observation group were more obvious than those in the control group (APACHEⅡ score: 10.29±1.86 vs. 15.35±2.06, SOFA score: 5.51±1.08 vs. 7.65±1.58, both P < 0.05). The length of ICU stay was shortened in the observation group than that in the control group (days: 12.22±9.48 vs. 20.22±15.35, P < 0.05). The incidence of MODS [35.0% (14/40) vs. 47.5% (19/40)] and the 28-day mortality [45.0% (18/40) vs. 47.5% (19/40)] was lower than that of the control group, but there was no statistical difference (both P > 0.05). Conclusion:Self-made Qingyuan Shenghua decoction can effectively improve the prognosis of patients with coagulation dysfunction and sepsis, and its mechanism may be related to inhibition of inflammatory reaction and improvement of coagulation function.