ABSTRACT
Objective:To investigate the effects of ursolic acid (UA) on proliferation, migration and iron death of ectopic endometrial stromal cells (EESCs) and its mechanism.Methods:Mouse model of endometriosis was established and the primary EESCs were isolated. The cells were treated with UA at different concentrations (0, 2.5, 5, 10, 20, 40, 50, 80, 100, 200 μmol/L). The cells were divided into Control group (normal culture), 2.5 μmol/L UA group (2.5 μmol/L UA treatment), 5.0 μmol/L UA group (5.0 μmol/L UA treatment), 10.0 μmol/L UA group (10 μmol/L UA treatment), and UA+DUSP19 group (10 μmol/L UA+50 μmol/L JAK2/STAT3 signal pathway activator DUSP19 treatment). Cell survival rate was detected by CCK-8 method. Cell proliferation was detected by plate cloning method. Transwell chamber assay was used to detect cell migration. The levels of Fe 2+ and the contents of malondialdehyde (MDA), reactive oxygen species (ROS) and superoxide dismutase (SOD) were detected by kit. Protein expression levels of Ki67, PCNA, CyclinD1, p-JAK2, p-STAT3, JAK2 and STAT3 were detected by western blot. Results:The number of clones in Control, 2.5 μmol/L UA, 5.0 μmol/L UA and 10.0 μmol/L UA groups were as follows: 152.22±15.47, 121.22±11.54, 92.00±5.54, 66.44±6.88; Ki67 protein expression was 1.08±0.10, 0.73±0.07, 0.61±0.06, 0.45±0.02, respectively; The expression of PCNA protein was 0.85±0.07, 0.64±0.05, 0.41±0.03, 0.31±0.05, respectively; CyclinD1 protein expression levels were 0.98±0.11, 0.65±0.06, 0.51±0.05, 0.42±0.07, respectively. The migration numbers were 92.78±6.27, 62.22±2.20, 50.22±4.59 and 39.11±4.33, respectively; Fe 2+ levels were (1.06±0.07) μmol/g, (1.21±0.11) μmol/g, (1.33±0.08) μmol/g, (1.47±0.09) μmol/g, respectively; MDA content was (0.48±0.06) μmol/g, (0.65±0.07) μmol/g, (0.85±0.08) μmol/g, (1.03±0.11) μmol/g, respectively; ROS contents were (19.85±1.21) %, (24.83±2.79) %, (29.04±1.86) %, (33.87±2.45) %, respectively; SOD content were (36.41±3.56) U/mg, (31.03±2.81) U/mg, (25.63±2.84) U/mg, (19.62±1.67) U/mg, respectively; p-JAK2 protein expression was 0.85±0.10, 0.75±0.06, 0.53±0.05, 0.31±0.03, respectively; p-STAT3 protein expression was 1.08±0.11, 0.79±0.06, 0.63±0.07, 0.42±0.03, respectively. The p-JAK2 protein content in UA group and UA+DUSP19 group was 0.38±0.05 and 0.75±0.08, respectively; p-STAT3 protein expression was 0.46±0.04 and 0.80±0.03, respectively; The cell survival rates were (52.55±2.44) % and (82.18±4.72) %, respectively; Fe 2+ levels were (1.57±0.06) μmol/g and (1.21±0.13) μmol/g, respectively. The differences in the above indicators between the Control group and the 2.5 μmol/L UA group, 5.0 μmol/L UA group and 10.0 μmol/L UA group were statistically significant ( P<0.05). There were statistically significant differences among 2.5 μmol/L UA group, 5.0 μmol/L UA group and 10.0 μmol/L UA group ( P<0.05). There were statistically significant differences in p-JAK2, p-STAT3, cell survival rate and Fe 2+ levels between UA group and UA+DUSP19 group ( P<0.05) . Conclusion:Ursolic acid can inhibit the proliferation and migration of EESCs cells and induce iron death by regulating JAK2/STAT3 signaling pathway, thus playing a protective role in endometriosis.
ABSTRACT
Objective:To investigate the effect of single nucleotide variation of osteoprotegerin (OPG) gene on the occurrence of osteoporosis (OP) in patients with gestational diabetes mellitus (GDM) .Methods:From Apr. 2018 to Apr. 2022, 276 pregnant women with GDM who underwent prenatal examination and gave birth in Linyi People’s Hospital were collected for analysis, general data were collected and bone mineral density was tested. According to the bone mineral density test results, they were divided into normal group and OP group. The OPG genotype was tested, and the general information, OPG genotype and allele frequency of the two groups were compared. The differences in bone mineral density among different genotypes of OPG were compared, and the genotypes affecting the risk of OP in GDM patients were analyzed.Results:There was no significant difference in the general data of the two groups of patients (all P>0.05). The allelic distribution of the rs3134069 and rs2073618 loci of the OPG gene in the two groups of patients conformed to the Hardy-Weinberg equilibrium law (all P>0.05). There was a statistically significant difference in the frequency of the AC genotype at rs3134069 between the two groups ( χ2=7.75, P=0.005). Taking patients with the AA genotype as a reference, patients with the AC genotype had a lower risk of developing OP ( OR=0.15, 95% CI: 0.03-0.59). There was a statistically significant difference in the frequency of CC genotype at rs2073618 between the two groups ( χ2=11.30, P=0.001). Taking patients with GG genotype as a reference, patients with CC genotype had a higher risk of developing OP ( OR=7.42, 95% CI: 2.19-27.18). Comparing rs3134069 and rs2073618 loci, there was no significant difference in bone mineral density at each part of the three genotypes (all P>0.05). The multivariate Logistic regression model showed that the AC genotype of rs3134069 ( OR=0.18, 95% CI: 0.03-0.70, P=0.029) was a protective factor for the induction of OP, while GC genotype of rs2073618 ( OR=6.86, 95% CI: 1.57-27.15, P=0.007) were the risk factors for OP in GDM patients. Conclusion:The CC genotype of rs2073618 is significantly positively correlated with the susceptibility to OP in GDM patients.
ABSTRACT
Objective:To analyze the clinical outcome of vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) in infertile patients with polycystic ovary syndrome (PCOS) combined with insulin resistance (IR) .Methods:A total of 257 PCOS infertile patients undergoing IVF/ICSI-ET from Jan. 2018 to Dec. 2020 were included and retrospectively analyzed. The patients were divided into IR group (HOMA-IR≥2.5, 130 cases) and non-IR group (HOMA-IR<2.5, 127 cases) according to the level (median 2.5) of homeostasis model assessment of insulin resistance index (HOMA-IR) . The levels of basic sex hormones [follicular stimulating hormone (FSH) , luteinizing hormone (LH) , estradiol (E2) , testosterone (T) , progestational hormone (P) , anti-mullerian hormone (AMH) ] and numbers of basic sinus follicles, levels of blood glucose and insulin at 30min, 60min and 120min after glucose administration and fasting and proconceptive pregnancy outcome indicators[gonadotropin (Gn) use time and dose, number of eggs obtained, fertilization rate, high-quality embryonic rate, occurrence rate of ovarian hyperstimulation syndrome (OHSS) , implantation rate, clinical pregnancy rate, biochemical pregnancy rate, abortion rate, live birth rate and pregnancy complications] were compared between the two groups. The influencing factors of clinical outcomes were analyzed by Logistic regression.Results:The levels of basic LH [ (8.86±1.60) mIU/ml vs (6.54±1.12) mIU/ml], T[ (63.20±7.47) ng/dl vs (52.11±5.69) ng/dl] in IR group was significantly higher than those in non-IR group ( P<0.05) . At different time-point, the levels of blood glucose and insulin in IR group were significantly higher than those in non-IR group ( P<0.05) . The Gn dose [ (1947.35±129.13) IU vs (1522.70±88.41) IU] and abortion rate [32.69% (17/52) vs 13.70% (10/73) ] in IR group was significantly higher than those in non-IR group ( P<0.05) , and the clinical pregnancy rate [40.00% (52/130) vs 57.48% (73/127) ] and live birth rate [51.92% (27/52) vs 72.60% (53/73) ] was significantly lower than those in non-IR group ( P<0.05) . Logistic regression analysis showed that age, BMI, basic LH, basic T and HOMA-IR was independent risk factors for clinical outcome of IVF/ICSI-ET in infertility patients with PCOS ( P<0.05) , and basic AMH and Gn dose were protective factors for clinical outcome ( P<0.05) . Conclusion:IR negatively affects the clinical outcome of IVF/ICSI-ET in infertile patients with PCOS, HOMA-IR is a risk factor for clinical outcomes, and IR should be evaluated in time for infertile patients with PCOS.
ABSTRACT
OBJECTIVE：To screen the effective compo nent in antioxi dant active fraction of Pueraria lobata . METHODS ：The antioxidant active fraction sample （S1-S20） of 20 batches of P. lobata were prepared. HPLC method was adopted. The determination was performed on SepaxBio-C 18 column with mobile phase consisted of methanol-water （gradient elution ）at the flow rate of 0.6 mL/min. The column temperature was set at 25 ℃，and detection wavelength was set at 250 nm. HPLC fingerprints of 20 batches of P. lobata were established by the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition），and common peaks were identified. Cluster analysis ，principal component analysis （PCA）and orthogonal partial least squares discriminant analysis （OPLS-DA）were used to screen the effective components in antioxidant active fraction of P. lobata . RESULTS：There were 18 common peaks in HPLC fingerprints of 20 batches of antioxidant active fraction in P. lobata ，and the similarity was more than 0.99. Eight common peaks were identified ，which were 3′-hydroxypuerarin（peak 2），puerarin（peak 3）， 3′-methoxypuerarin（peak 4），daidzein（peak 5），genistein（peak 7），formononetin（peak 11），daidzein（peak 13）and genistein （peak 16）. The results of cluster analysis and PCA analysis showed that samples S 1，S3，S4，S6，S8，S18 and S 19 were clustered into one category ，and samples S 2，S5，S7，S9-S17 and S 20 were clustered into one category ；peak 2，peak 3，peak 10，peak 11 and peak 13 had great influence on principal component 1；peak 8 and peak 9 had great influence on principal component 2. OPLS-DA analysis showed that peak 4，peak 3，peak 2，peak 16，peak 13 and peak 11 had great influence on the quality of antioxidant active fraction of P. lobata . CONCLUSIONS ： HPLC fingerprint for active fraction of P. lobata is established in the study and 8 components are identified ；among them ， com puerarin，3′-hydroxypuerarin，daidzein and formononetin maybe the material basis of antioxidant fraction of P. lobata .
ABSTRACT
OBJECTIVE：To optimi ze the optimal composition proportion of 4 ingredients （Panax ginseng ，Astragalus membranaceus，Polygonatum sibiricum ，Lycium chinensis ）in Compound ginseng immune-enhancing formula （CGIF），and to study immune activity and acute toxicity of the extracts with the optimal ratio. METHODS ：The cell activity test was used to screen the crude drug concentration range of 4 ingredients. After treated with different crude drug concentrations of each medicinal material，using the contents of NO ，IL-6 and TNF-α as indexes，uniform design was used to determine the optimal ratio of each ingredient in CGIF. Totally 240 mice were taken and randomly divided into 4 batches，with 60 mice in each batch. Each batch of mice was randomly divided into blank group （normal saline ），model group （normal saline ），positive drug group [levamisole ，4 mg/（kg·d）]，and optimal proportion extract of CGIF low-dose ，medium-dose and high-dose groups [ 0.952 8，1.905 6，3.811 2 g/（kg·d）]，with 10 mice in each group ；they were given medicine intragastrically ，qd，for consecutive 30 d. Except for blank group，mice in the other groups were intraperitoneally injected with cyclophosphamide [ 40 mg/（kg·d）] on the 24th day after first administration，qd，for consecutive 3 d to induce immunocompromised model. The immune activity of the optimal proportion extract was evaluated by determining visceral coefficients ，spleen lymphocyte transformation capacity ，serum contents of hemolysin，IL-2，IgM，IgG and IgA ，phagocytosis function of peritoneal macrophages. Another 20 mice were collected and given the optimal proportion extract 20 mL/kg intragastrically ，twice；acute toxicity of the formula was investigated with oral maximum tolerated dose （MTD）. RESULTS ：The optimal ratio of CGIF was that crude drug mass ratio of P. ginseng ， membranaceus，P. sibiricum ，L. chinensis was 1 ∶ 2 ∶ 2 ∶ 4. The immunological activity experiment showed that theoptimal proportion extract can significantly improve visceral indexes of mice ， spleen lymphocyte proliferation ability serum contents of hemolysin ，IL-2，IgM，IgG and IgA as well as macrophage phagocy tosis ability （P＜0.05 or P＜ 0.01）. The acute toxicity test indicated that oral MTD was over 15 g/kg，which was non-toxic. CONCLUSIONS ：The optimal proportion extract of CGIF can significantly enhance the immune function of mice and are non-toxic.
ABSTRACT
Antimicrobial peptides (AMPs) are small molecules produced by a myriad of cells and play important roles not only in protecting against infections and sustaining skin barrier homeostasis but also in contributing to immune dysregulation under pathological conditions. Recently, increasing evidence has indicated that AMPs, including cathelicidin (LL-37), human β-defensins, S100 proteins, lipocalin 2, and RNase 7, are highly expressed in psoriatic skin lesions. These peptides broadly regulate immunity by interacting with various immune cells and linking innate and adaptive immune responses during the progression of psoriasis. In this review, we summarize the recent findings regarding AMPs in the pathogenesis of psoriasis with a main focus on their immunomodulatory abilities.
Subject(s)
Humans , Adaptive Immunity , Immunity, Innate , Pore Forming Cytotoxic Proteins , Psoriasis , Skin Diseases , beta-DefensinsABSTRACT
OBJECTIVE：To optimi ze the ratio of four comp onents of Compound renshen jianti formulation （Panax ginseng ， Dioscorea oppositifolia ，Lycium barbarum fruit，Alpinia oxyphylla ），and to investigate its anti-fatigue activity and acute toxicity. METHODS：The water extract of Compound renshen jianti formulation was prepared by water extraction ，concentration and decompression drying. By single factor tests ，using weight-bearing swimming time as index ，the effects of four factors were investigated，such as the amount of P. ginseng ，D. oppositifolia ，L. barbarum fruit，A. oxyphylla . On the basis of single factor tests，using comprehensive score of weight-bearing swimming time ，serum urea nitrogen content ，liver glycogen content and AUC of blood lactate after exercise as index ，the formulation was optimized by Box-Behnken response surface method. The mice was divided into blank control group （water），positive control group （Renshen hongjingtian capsules ，0.135 g/kg）and compound low-dose，medium-dose and high-dose groups [the optimal ratio of Compound renshen jianti formulation extract （called“optimal compound formulation ”for short ）4.08，8.16，12.24 g/kg，by crude drug] ，intragastric administration of drug or distilled water 20 mL/kg，once a day ，for consecutive 30 d. The weight-bearing swimming time ，the contents of serum urea nitrogen ，liver glycogen and blood lactate AUC after exercise were used to optimize its anti-fatigue activity of optimal compound formulation. The comprehensive score was calculated based on the measured data of mice in the compound formulation middle-dose group ， and the difference between it and the theoretical prediction value was compared. The mice were given optimal compound formulation intragastrically （total dose 16.00 g/kg， by extract）. The general state ， body mass change ， toxic characteristics and death of mice were observed and recorded for 14 days. Median lethal dose （LD50）and maximum tolerated dose （MTD）were measured. RESULTS ：The optimal formulation ratio of Compound renshen jianti formulation included that P. ginseng 1.5 g，D. oppositifolia 10 g，L. barbarum fruit 10 g，A. oxyphylla 3 g. Results of anti-fatigue activity validation test showed that the optimal compound formulation could significantly prolonged weight-bearing swimming time ，reduced serum content of urea nitrogen ，blood lactate content and its AUC （except for low-dose group ），while significantly increased the content of liver glycogen （P＜0.05 or P＜0.01）. Average comprehensive score of medium-dose group was 96.95，which was only 0.06％ different from the theoretical prediction value of 97.01. The results of acute toxicity test showed that there was no death in mice. The oral MTD of the optimal compound formulation was more than 15 g/kg，which was non-toxic. CONCLUSIONS ：The optimal Compound renshen jianti formulation has effective anti-fatigue activity of mice ，and has no significant toxic effect.
ABSTRACT
Objective:To investigate the role of neutrophil extracellular traps (NETs) in psoriatic lesions in activation of absent in melanoma 2 (AIM2) inflammasomes in keratinocytes.Methods:Four skin specimens and 4 peripheral blood specimens were collected from patients with advanced psoriasis vulgaris, who were treated at Department of Dermatology, Xijing Hospital, the Fourth Military Medical University from January to December in 2018. In addition, 4 skin specimens were collected from healthy human controls, and 3 foreskin specimens from children aged under 15 years after circumcision. Tissue immunofluorescence study was performed to determine the expression of NETs and AIM2 inflammasomes in normal skin tissues and psoriatic lesions. Neutrophils were separated from peripheral blood of patients with psoriasis vulgaris by using magnetic beads, and NETs were extracted. Primary keratinocytes were isolated from foreskin tissues, and divided into 4 groups to be stimulated with phosphate-buffered saline (PBS) (control group) , NET extracts (NET group) , DNase Ⅰ-treated NET extracts (NET degradation group) or DNase Ⅰ (degrader control group) respectively for 48 hours. Western blot analysis was performed to determine the expression of AIM2 inflammasomes and its downstream molecules, and enzyme-linked immunosorbent assay (ELISA) to detect the level of IL-1β in the cell culture supernatant in the NET group and control group. Statistical analysis was carried out by using one-way analysis of variance and Dunnett- t test for multiple comparisons. Results:NET structures were observed in the epidermis of psoriatic lesions, but not in that of the healthy controls. Besides, the expression of AIM2 inflammasomes was higher in the epidermis of psoriatic lesions than in the healthy controls. Western blot analysis showed that there were significant differences in the protein expression of AIM2 and its downstream molecules pro-IL-1β and IL-1β among groups ( F = 23.80, 5.82, 15.64 respectively, all P < 0.001) . The NET group showed significantly higher protein expression of AIM2 (1.42 ± 0.03) , pro-IL-1β (1.32 ± 0.08) and IL-1β (1.40 ± 0.05) compared with the control group ( t = 15.14, 4.26, 8.71, respectively, all P < 0.05) , while no significant difference in the expression of AIM2, pro-IL-1β and IL-1β was observed between the NET degradation group (1.15 ± 0.07, 0.93 ± 0.03, 1.07 ± 0.05, respectively) and control group ( t = 2.10, 2.18, 1.40 respectively, all P > 0.05) . In addition, the IL-1β level in the cell culture supernatant was significantly higher in the NET group (13.15 ± 3.77 pg/ml) than in the control group (3.61 ± 0.20 pg/ml, t = 2.53, P < 0.05) . Conclusion:NETs exist in the epidermis of psoriatic lesions, can aggravate the inflammatory process in psoriasis, and contribute to the occurrence and development of psoriasis, likely by activating AIM2 and promoting the cleavage and secretion of IL-1β in keratinocytes.
ABSTRACT
Objective:To investigate the mechanism of Three Handing-Three Points on pain function in sciatic nerve injury rats by observing the changes of chemokine (C-X3-C motif) ligand 1, CX3CL1)/chemokine (C-X3-C motif) receptor 1 (CX3CR1) protein and mRNA expression in spinal dorsal horn. Methods:A total of 74 male Sprague-Dawley rats were randomly divided into normal group (n= 12), sham group (n = 24), model group (n = 25), and Three Handing-Three Points group (Tuina group,n = 13). The model group and Tuina group prepared the sciatic nerve injury model. The sham group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with Tuina manipulation emulator. The photothermal pain threshold was measured seven days after modeling and after 20 days of intervention; cumulative pain score was measured seven days after modeling, and after ten days and 20 days of intervention. The spinal dorsal horn tissues were extracted to detect the protein and mRNA expression of CX3CL1/CX3CR1 with Western blotting and RT-PCR seven days after modeling and after 20 days of intervention. The microglia morphology in spinal dorsal horn was observed with immunofluorescence after 20 days of intervention. Results:Seven days after modeling, compared with the normal group, the photothermal pain tolerance threshold increased in the model group and the sham group (P < 0.05); compared with the sham group, the cumulative pain score increased in the model group and Tuina group (P < 0.05). After ten days of intervention, the cumulative pain score was lower in Tuina group than in the model group (P < 0.05). After 20 days of intervention, both the photothermal pain tolerance threshold and cumulative pain score were lower in Tuina group than in the model group (P < 0.05). There was no significant difference in the expression of CX3CL1/CX3CR1 protein and mRNA on the seven days after modeling and after 20 days of intervention (P > 0.05). The microglia in the model group were partially activated or completely activated, while those in Tuina group were unactivated or partially activated after 20 days of intervention. Conclusion:Three Handing-Three Points can improve the pain function of sciatic nerve injured rats, which may associate with regulating microglia through the pathway other than CX3CL1/CX3CR1.
ABSTRACT
Objective:To investigate the effect of Tuina of Three Handing-Three Points on the motor function of hind limbs, the proliferation of Schwann cell, recovery of myelin sheath and the expression of transforming growth factor (TGF)-β1/Smad2 pathway protein in injured sciatic nerve of rats. Methods:A total of 66 male Sprague-Dawley rats were randomly divided into sham operation group (n = 22), model group (n = 22) and observation group (n = 22). The sciatic nerve injury model was made by clamping method. On the eighth day after modeling, the observation group received stimulation on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34). The sciatic functional index (SFI) was measured before intervention and 21 days after intervention. The Oblique Plate Test was performed before intervention, and seven days, 14 days and 21 days after intervention. The expression of S100, TGF-β1 and Smad2 were observed by immunofluorescence. The expression of TGF-β1, Smad2 and p-Smad2 was detected by Western blotting. Results:Before intervention, SFI was lower in the model group and observation group than in the sham operation group (P < 0.05); 21 days after intervention, SFI and the angle of Oblique Plate Test were higher in the observation group than in the model group (P < 0.05). Immunofluorescence showed that, 21 days after intervention, the expression of S100 was lower in the model group than in the sham operation group (P < 0.01), and was higher in the observation group than in the model group (P < 0.05), no difference was found between the observation group and the sham operation group (P > 0.05). Western blotting showed that, before intervention and seven days after intervention, the expression of TGF-β1, Smad2 and p-Smad2 were higher in the model group than in the sham operation group; 21 days after intervention, no difference was found in the expression among groups (P > 0.05) Conclusion:Three Handing-Three Points could promote the proliferation of Schwann cell and the recovery of myelin sheath, to improve the motor function of hind limbs in rats with sciatic nerve injury, which may not be related to TGF-β1/Smad2 pathway.
ABSTRACT
Objective:To explore the effect of Tuina of Three Handing-Three Points on the recovery of motor function, the expression of neuregulin (NRG) 1 and human epidermal growth factor receptor (ErbB) 2 in the injured point of sciatic nerve and L4-6 spinal cord, and the morphological change of myelin sheath at the injured point of sciatic nerve of rats. Methods:A total of 76 male Sprague-Dawley rats were randomly divided into normal group, sham operation group, model group and Tuina group with 19 rats in each group. The right side sciatic nerve was clamped to make model in the model group and Tuina group. The sham operation group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with dialing, plucking and kneading using Tuina technique simulator. All of them were tested with Oblique Plate Test before modeling, seven days and 28 days after modeling. Western blotting was used to detect the protein expression of NRG1 and ErbB2 in the injured point of sciatic nerve and L4-6 spinal cord. The change of myelin sheath at the sciatic nerve injury point was observed and analyzed by transmission electron microscope. Results:Seven days and 28 days after modeling, the scores of Oblique Plate Test were lower in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the scores was higher in Tuina group than in the model group (P < 0.05). At the sciatic nerve injury point, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); seven days and 28 days after modeling, there was no significant difference in NRG1 among groups (P > 0.05); 28 days after modeling, the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05). In L4-6 spinal cord, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and sham operation group (P < 0.05); seven days after modeling, the expression of NRG1 was higher in the model group and Tuina group than in the sham operation group (P < 0.05), and the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the expression of NRG1 was higher in Tuina group than in the model group (P < 0.05), and there was no significant difference in ErbB2 among groups (P > 0.05). The electron microscope showed that, 28 days after modeling, the myelin sheath collapsed seriously in the model group; while the ultrastructure of the nerve injury point improved, and the myelin sheath of the nerve fiber was relatively intact in Tuina group; the g-ratio value was lower in the model group than in the sham operation group (P < 0.05), the g-ratio value was higher in Tuina group than in the model group (P < 0.05), and no difference was found in g-ratio value between Tuina group and sham operation group (P > 0.05). Conclusion:Three Handing-Three Points could improve the motor function of hind limbs in rats with sciatic nerve injury, which may be related to the adjustment of NRG1 and ErbB2 in the sciatic nerve and spinal cord, to maintain normal myelin sheath structure.
ABSTRACT
Objective: To study the effect of Rosae Chinensis Flos total flavones(RCTF) on the focal cerebral ischemia-reperfusion model in rats, in order to preliminarily explore the mechanism of action. Method: Rats were randomly divided into sham-operated group, model group, large, medium, and low-dose RCTF group(200,100,50 mg ·kg-1) and positive group[Nimodipine group(20 mg ·kg-1) and Naoluotong group (500 mg ·kg-1)]. After 7 days of continuous administration, 1 hour later after the last administration, the middle cerebral artery middle cerebral artery occlusion (MCAO) model was duplicated. After 2 hours of modeling, perfusion was performed for 22 hours. Mortality and neurological deficits were scored. Serum S-100β was detected; brain tissue malondialdehyde(MDA), superoxide dismutase (SOD), nitric oxide (NO), nitric oxide synthase (NOS), tumour necrosis factor-α(TNF-α), interleukin-1β(IL-1β), intercellular adhesion molecule-1(ICAM-1), adenosine triphosphate (ATP)ase were measured. The brain tissue morphological changes were observed. Result: The rat model of focal cerebral ischemia and reperfusion was successfully replicated. Compared with the model group, RCTF in large, medium, and low-dose RCTF group significantly decreased the score of neurological deficit in rats (Pβ in serum (PPP+K+-ATPase, Mg2+-ATPase, and Ca2+ in brain tissue (Pα content, IL-1β, ICAM-1 content in brain tissue (PPConclusion: RCTF have a protective effect on cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the resistance of anti-free radicals, the reduction of inflammation in brain tissue and the improvement of brain energy metabolism after cerebral ischemia reperfusion injury.
ABSTRACT
As the primary innate immune cells in the central nervous system, microglia can be activated by external noxious stimulus and in turn interact with astroglia and neurons to induce neuroinflammation and facilitate the transmission of pain signals. This response can help the central nervous system adapt to the changes of the internal environment induced by noxious stimulus, leading to the long-term sensitivity of peripheral and central pain nerve conduction pathways and chronic neuropathic pain. Numerous researches found that activation of microglia participated in the occurrence and maintenance of chronic neuropathic pain, and inhibition of microglial activation in the spinal cord or the brain had analgesic effect in animal experiments. Due to the fact that molecular and cellular mechanisms between the activation of microglia and pain remittence are unclear, there are many difficulties in designing of new drugs selectively targeting to the activation of microglia for treatment of chronic neuropathic pain. We review here the research articles on microglia and chronic neuropathic pain, sorting out the relationship between microglia and chronic neuropathic pain, and provide new ideas for the development of new drugs targeting to microglia for the treatment of chronic neuropathic pain.
ABSTRACT
Objective To observe the time dependent changes of pancreas and lung injury in mice acute pancrea-titis induced by caerulein,and compare the effects of mice acute pancreatitis model induced by different injection protocols of caerulein and lipopolysaccharides. Methods Acute pancreatitis were induced by different injection frequency of caerulein or caerulein combined with lipopolysaccharides, and plasma amylase activity was detected by starch-iodine colorimetry,and pancreas and lung injury severity was observed on paraffin section after HE stai-ning. Results The injury of pancreas and lung in mice acute pancreatitis induced by caerulein was most severe at 4 h, which began to repair at 24 h, and basically returned to normal at 5 d. 4 h after last injection, the amylase activity in CAE7 group and CAE9 group was higher than that in the control group(6 461±1 078 U/dl vs 3 093± 331 U/dl;6 821 ± 495 U/dl vs 3 093 ± 331 U/dl, all P<0.001), and CAE7+LPS group was higher than in CAE9 group[(8 912±465)U/dl vs (6 821±495)U/dl,P<0.001],the histological score in CAE7 group and CAE9 group was higher than that in the control group(4.750 ± 0.524 vs 0 ± 0; 4.917 ± 0.664 vs 0 ± 0, all P<0.001), and that in CAE7+LPS group was higher than in CAE9 group(7.167 ± 0.258 vs 4.917 ± 0.664, P<0.001). Conclusions Mice acute pancreatitis can be successfully induced by 7 i.p. injections of caerulein (50 μg/kg body weight) at 1-hour intervals,caerulein combined with lipopolysaccharides.
ABSTRACT
Objective To observe the time dependent changes of pancreas and lung injury in mice acute pancrea-titis induced by caerulein,and compare the effects of mice acute pancreatitis model induced by different injection protocols of caerulein and lipopolysaccharides. Methods Acute pancreatitis were induced by different injection frequency of caerulein or caerulein combined with lipopolysaccharides, and plasma amylase activity was detected by starch-iodine colorimetry,and pancreas and lung injury severity was observed on paraffin section after HE stai-ning. Results The injury of pancreas and lung in mice acute pancreatitis induced by caerulein was most severe at 4 h, which began to repair at 24 h, and basically returned to normal at 5 d. 4 h after last injection, the amylase activity in CAE7 group and CAE9 group was higher than that in the control group(6 461±1 078 U/dl vs 3 093± 331 U/dl;6 821 ± 495 U/dl vs 3 093 ± 331 U/dl, all P<0.001), and CAE7+LPS group was higher than in CAE9 group[(8 912±465)U/dl vs (6 821±495)U/dl,P<0.001],the histological score in CAE7 group and CAE9 group was higher than that in the control group(4.750 ± 0.524 vs 0 ± 0; 4.917 ± 0.664 vs 0 ± 0, all P<0.001), and that in CAE7+LPS group was higher than in CAE9 group(7.167 ± 0.258 vs 4.917 ± 0.664, P<0.001). Conclusions Mice acute pancreatitis can be successfully induced by 7 i.p. injections of caerulein (50 μg/kg body weight) at 1-hour intervals,caerulein combined with lipopolysaccharides.
ABSTRACT
Objective To observe the effect and the mechanisms of ferulic acid on radiationinduced damage of mice peripheral blood and bone marrow hematopoietic function.Methods Ninety-six mice were randomly divided into sham irradiation group,irradiation group,positive drug group and 10,30,90 mg·kg-1 ·d-1 ferulic acid group,16 mice per group.Mice were exposed to 3.5 Gy γ-rays 24 h after first drug taken.Then,mice were given drugs for 7 d after irradiation.White blood cells in peripheral blood of 10 mice per group were counted 2 d before irradiation and 3,7,10,15 and 22 days after irradiation.The bone narrow of the other six mice was taken to detect the micronuclei frequency of polychromatic erythrocyte,the hematopoietic progenitor cell colony formation capacity,Thbd and HMGB1 protein expressions in mice bone marrow on the seventh day after irradiation.Results Compared with the irradiation alone group,the treatment of mice with ferulic acid 90 mg· kg-1 · d-1 increased the number of white blood cells in peripheral blood at 3,10,15 and 22 d after irradiation (t =2.267,2.399,1.945,2.828,P ＜ 0.05).Treatment with mice with ferulic acid 90 mg· kg-1 · d 1 decreased the micronuclei rate of erythrocytes in irradiated bone marrow (t =4.013,P ＜ 0.05),increased the clone numbers of CFU-E,BFU-E and CFU-GM of hematopoietic progenitor cells (t =2.366,2.953,3.115,P ＜0.05),improved the relative expression of the Thbd protein in bone marrow and the HMGB1 protein in nuclear (t =17.75,23.39,P ＜ 0.01).Conclusions Ferulilc acid could protect the bone marrow hematopoietic of mice exposed to irradiation by regulating the expressions of Thbd and HMGB1 protein,and then accelerate the peripheral cells recovery.
ABSTRACT
OBJECTIVE: To observe the in vivo metabolism and distribution characteristics of nano-cerium oxide( nanoCeO_2) in rats,and to explore the radio-protective effect of nano-CeO_2. METHODS: i) A total of 18 specific pathogen free( SPF) SD rats were randomly divided into 3 groups. Rats of experiment group and CeO_2 blood group were gavaged with1. 0 g/kg body weight( bw) nano-CeO_2 suspension. Rats of control group were gavaged with double distilled water( DDW)in equal volume. At different time-points after treatment,venous blood was collected from the rats' eye socket in CeO_2 blood group,meanwhile urine and excrement of rats of experiment group were also collected. Organ and tissue samples of experiment group and control group were collected 24. 0 hours after treatment. The concentrations of cerium in biological samples were detected by inductively coupled plasma mass spectrometry. ii) A total of 72 SPF BALB/c mice were randomly divided into 6 groups. Mice of low-,medium-and high-dose groups were gavaged with 100,300 and 900 mg/kg bw nano-CeO_2 suspension respectively. Mice of negative control group,irradiation control group and drug positive control group were gavaged with DDW in equal volume once daily. After 14 days,mice of the other 5 groups were exposed by60Coγ-rays once with 3. 5 Gy( 1 Gy/min) except the negative control group. Mice of drug positive control group were given intraperitoneal injection with 200 mg/kg bw amifostine half an hour before irradiation. After exposure,mice were treated by the above gavages once daily. After 3 and 8 days,6 mice were randomly selected to collect the peripheral blood for the count of white blood cell( WBC) and lymph cell measuring. RESULTS: i) The cerium concentration in blood reached peak value in 4. 0 hours after exposure of nano-CeO_2,and the cerium concentration of urine and excrement reached maximum in8. 0 hours after exposure. After 24. 0 hours of exposure,the cerium concentration of brain in experiment group was higher than that of control group( P < 0. 05). Among the experiment group,the cerium concentrations of sternum,duodenum and brain were higher than that of kidney and heart( P < 0. 05),meanwhile the cerium concentrations of thymus and lung were higher than that of kidney( P < 0. 05). ii) There was no statistical difference in interactive effect of WBC count and lymph cell counts between nano-CeO_2 exposure ways and time( P > 0. 05). The WBC counts of the low-and medium-dose groups were lower than those of the negative control group and the drug positive control group( P < 0. 05). The WBC count of high-dose group was lower than those of irradiation control group,drug positive control group and medium-dose group( P <0. 05). The lymph cell counts of the 3 dose groups were lower than that of drug positive control group( P < 0. 05).CONCLUSION: The nano-CeO_2 is mainly cumulated in organs such as sternum,duodenum,brain,thymus and lung. After induced by radiation,nano-CeO_2 has a certain degree of promotion role in increasing the WBC counts.
ABSTRACT
[Abstact] Objective To study the preformulation properties of tecovirimat for formulation design. Methods The appear?ance,crystal structure,solubility and permeability of the drug were investigated. The UV method was established to determine the con?tent of tecovirimat in vitro. The solubilization experiment was also conducted. Results Tecovirimat is white and odorless powder with crystalline hydrate structure and low water-solubility with high permeability. The morphology of tecovirimat is six-prismatic-shape. The linearity range of established UV method was 4.14-24.83μg/ml(r=0.9996). The 1∶1 soluble complex was formed with tecovirimat and hydroxypropyl-β-cyclodextrin. Conclusion Tecovirimat is poorly water-soluble drug with high permeability and the established meth?od could be used to determine the content of the drug. Hydroxypropyl-β-cyclodextrin could be used for the solubilization of tecovirimat.
ABSTRACT
BACKGROUND:Mesenchymal stem cels isolated from cord blood and bone marrow have multi-directional differentiation ability under a certain condition of induction. OBJECTIVE:To compare the difference of differentiation of umbilical cord blood and bone marrow mesenchymal stem cels into osteoblasts. METHODS:Human umbilical cord blood and bone marrow mesenchymal stem cels were isolated and cultured by density gradient method. When reached 90% confluency, mesenchymal stem cels were digested by trypsin for subculture. At the third passage, umbilical cord blood mesenchymal stem cels and bone marrow mesenchymal stem cels at 8×104/wel were incubated. When reached 80% confluency, cels were treated with low-glucose DMEM supplemented with 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C and 10 mmol/L β-sodium glycerophosphate. RESULTS AND CONCLUSION:There was no significant difference in morphology and biological properties of the two kinds of mesenchymal stem cels. Cels were highly expressed CD44, CD29, but did not express CD34. They had the ability to differentiate into osteoblasts, which had a positive staining for known markers: alkaline phospatase and calciumin vitro mineralization. There was no significant difference in the activity of osteoblasts of two kinds of cels. Results verify that umbilical cord blood and adult bone marrow mesenchymal stem cels can be induced into osteoblasts with a similar ability, and they can be used as seed cels for bone tissue engineering.
ABSTRACT
Objective This study aimed to optimize the preparation condition of DNA-chitosan nanoparticles with high transfec-tion efficency through a central composition design .Methods The DNA-chitosan nanoparticles were prepared by complex coacervation between pEGFP and chitosan .We selected the concentrations of chitosan and plasmid as two experimental factors , and a central compos-ite design with two factors and five levels was used to optimize the preparation condition of DNA-chitosan nanoparticles for high transfec-tion efficency .The concentrations of chitosan and plasmid were selected as the independent variables , respectively .The dependent varia-bles included average particle size and transfection efficiency .The morphology of DNA-chitosan nanoparticles was observed using a trans-mission electron microscope .The size and zeta potential of nanoparticles were measured by dynamic light scattering ( DLS) and electro-phoretic light scattering ( ELS ) , respectively .The stability of plasmids in the process of nanoparticles preparation was investigated through the agrose gel electrophoresis .The expression of plasmids delivered by nanoparticles was observed under an inverted fluorescence microscope .The transfection efficiency of DNA-chitosan nanoparticles was assayed by flow cytometry .Results The preparation condition of DNA-chitosan nanoparticles with high transfection efficency was optimized successfully .Under the optimum preparative conditions , the DNA-chitosan nanoparticles were almost spherical .The average size of nanoparticles was 217.6nm, and distributed in a narrow range with a polydispersity index of 0.241.The zeta potential was +22.4 mV, which suggested that a den-sity of positive charge exist onto the surface of nanoparticles and consequently enhanced the stability of nanoparticles suspension .The results of gel electrophoresis showed that plasmids were not destroyed in the process of nanoparticles preparation .The cell transfection of nanoparticles was very highly efficient .The nanoparticles could effectively deliver the pEGFP plasmids into cells to express the green fluorescent protein at a high level.Conclusion The established mathematic models have the good predictive function .Under the optimum preparative condi-tions, the DNA-chitosan nanoparticles have the high potential of cell transfection .