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Objective:To investigate the effects of fasudil hydrochloride(FH) on Rho-associated kinase 2(ROCK2) protein and ferroptosis in hippocampal area during early brain injury in rats with subarachnoid hemorrhage(SAH).Methods:Total 36 SPF grade Sprague-Dawley rats were divided into 3 groups by random number table method: Sham group, SAH group and SAH+ FH (a ROCK2 protein inhibitor) group (FH goup) with 12 rats in each group.SAH animal model was established by internal carotid artery perforation.The rats in FH group were injected intraperitoneally with FH(15 mg/kg) 30 minutes after successful modeling, and rats in Sham group and SAH group were injected intraperitoneally with the same volume of 0.9% sodium chloride solution.Twenty-four hours after the intervention, shuttle box test was used to observe the learning and memory ability of rats.The Fe 2+ content in rat hippocampus tissue was detected by colorimetry, and the protein levels of ROCK2 and ferroptosis-related long-chain acyl-CoA synthetase 4(ACSL4) and glutathione peroxidase 4(GPX4) in hippocampus were detected by immunohistochemistry and Western blot.Statistical analysis was performed by SPSS 20.0 software.One-way ANOVA was used for multigroup comparison, and LSD test was used for further pairwise comparison. Results:(1)In the shuttle box test, there were statistically significant differences in the number of avoidance reactions and avoidance reaction time of rats among the three groups( F=20.348, 22.316, both P<0.05). The number of avoidance reaction in SAH group was less than that in Sham group ((17.92±2.94) times, (27.13±3.48) times, P<0.05), the time of avoidance reaction in SAH group was longer than that in Sham group ((9.15±2.87) s, (3.68±1.09) s, P<0.05), while the number of avoidance reaction in FH group ((21.63±4.11) times) was more than that in SAH group, and the time of avoidance reaction ((6.08±1.76) s) was shorter than that in SAH group (both P<0.05). (2) The colorimetry results showed that there was a statistically significant difference in the content of Fe 2+ in hippocampus of rats among the three groups( F=7.965, P<0.05). The Fe 2+ content in SAH group was significantly higher than that of Sham group((0.091±0.032) nmol/mg, (0.038±0.024) nmol/mg, P<0.05), and the Fe 2+ content in the FH group ((0.065±0.021) nmol/mg) was lower than that of SAH group ( P<0.05). (3) There were significant differences in the number of ROCK2, ACSL4 and GPX4 positive cells in hippocampus of rats among the three groups in immunohistochemistry ( F=7.602, 14.171, 36.077, all P<0.05). The positive cells of ROCK2 and ACSL4 in SAH group ((21.63±4.72), (55.13±19.41)) were significantly higher than those of Sham group ((11.63±3.62), (23.38±3.74)) (both P<0.05), and the positive cells of ROCK2 and ACSL4 in FH group ((15.88±6.64), (44.75±8.29)) were both lower than those of SAH group(both P<0.05), while the number of GPX4 positive cells in SAH group (25.38±6.30) was significantly lower than that of Sham group (60.25±10.36) ( P<0.05), and the number of GPX4 positive cells in FH group (45.13±7.51) was higher than that of SAH group( P<0.05). (4)The results of Western blot showed that there were significant differences in the expression levels of ROCK2, ACSL4 and GPX4 proteins in the hippocampus of rats among the three groups( F=4.812, 12.573, 10.849, all P<0.05). The protein expression levels of ROCK2 and ACSL4 in SAH group were significantly higher than those in Sham group(both P<0.05), and the protein expression levels of ROCK2 and ACSL4 in FH group were lower than those in SAH group (both P<0.05), while the expression level of GPX4 protein in SAH group (0.27±0.09) was significantly lower than that in Sham group( P<0.05), and the expression level of GPX4 protein in FH group was higher than that of SAH group ( P<0.05). Conclusion:FH can inhibit ferroptosis in the hippocampus and improve the learning and memory ability of rats, and the mechanism may be related with down-regulation of ROCK2 protein.
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Objective:To evaluate the effects of berberine on necroptosis of non-alcoholic fatty liver disease in mice and its relationship with adenosine monophosphate-activated protein kinase(AMPK)/ signal transducer and activator of transcription 6(STAT6) pathway.Methods:Twenty-five 8-week-old male C57BL/6N mice were divided into control group, steatotic liver group, berberine treatment group(200 mg·kg -1·d -1), AMPK inhibitor Compound C treatment group(0.2 mg·kg -1·d -1), and STAT6 inhibitor AS1517499 treatment group(10 mg·kg -1·d -1). After 12 weeks of intervention, the mice and liver tissue were weighed, and serum aspartate aminotransferase(AST), alanine aminotransferase(ALT), triglyceride, tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) as well as liver malondialdehyde and superoxide dismutase were measured; liver tissue HE, Masson, and oil red O staining were performed. Western blotting was used to detect the expressions of necroptosis related proteins[receptor interaction protein kinase 3(RIPK3), phosphorylated(p-) mixed lineage kinase domain-like(MLKL)], AMPK, p-AMPK, and p-STAT6. Results:Compared with control group, the steatotic liver group had higher quality of liver and liver index, and higher levels of serum AST, ALT, triglyceride, TNF-α, IL-1β, and oxidative stress( P<0.05); Liver tissue was full of cavity changes and inflammatory cell infiltration, widely distributed red lipid droplets and obvious blue fiber dyeing; The expressions of RIPK3 and p-MLKL were up-regulated ( P<0.05), but the levels of p-AMPK and p-STAT6 were relatively reduced ( P<0.05). Compared with the steatotic liver group, berberine intervention decreased liver quality and liver index, improved liver function, reduced blood lipid levels, pro-inflammatory factor expression and oxidative stress level, and significantly alleviated the degree of liver steatosis and fibrosis, the levels of RIPK3 and p-MLKL ( P<0.05), while the expressions of p-AMPK and p-STAT6 were increased significantly ( P<0.05). As compared with the berberine treatment, AMPK and STAT6 inhibitor treatment could offset the protective effect of berberine on steatotic liver, moreover, the expressions of RIPK3 and p-MLKL were increased ( P<0.05). There was no statistical difference in AMPK total protein content among the five groups ( P>0.05). Conclusion:Berberine can activate AMPK/STAT6 pathway to inhibit the necroptosis of hepatocyte, thus plays a protective role on non-alcoholic fatty liver disease in mice.
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Objective:To evaluate the role of silencing information regulator 1 (SIRT1)/nuclear factors E2-related factor2 (Nrf2) signaling pathway in berberine-induced reduction of renal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), renal I/R group (RIR group), berberine+ I/R group (B group), berberine+ I/R+ SIRT1 inhibitor EX527 group (BE group) and berberine+ I/R+ Nrf2 inhibitor ATRA group (BA group). After the right kidney was removed, the left renal artery was clamped for 45 min followed by reperfusion to establish the model of renal I/R injury.In B, BE, and BA groups, berberine 100 mg·kg -1·d -1 was given for intragastric administration at 14 days before surgery.In group BE and group BA, EX527 5 mg·kg -1·d -1 and ATRA 10 mg·kg -1·d -1 were injected intraperitoneally at 3 days before surgery, respectively.The equal volume of normal saline was given for 14 consecutive days in group S and group RIR.Blood samples were collected from orbital vein at 24 h of reperfusion for measurement of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations, for determination of the interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents (by enzyme-linked immunosorbent assay) and expression of SIRT1, Nrf2, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, nucleotide-binding oligomerization domain-like receptor containing pyrin domain (NLRP3) (by Western blot) and for examination of the pathological changes of renal tubules (with a light microscope). The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly increased in RIR, B, BE and BA groups, the expression of SIRT1, Nrf2, ASC, caspase-1 and NLRP3 was up-regulated in RIR, BE and BA groups, and the expression of SIRT1, Nrf2, caspase-1 and NLRP3 was up-regulated in group B ( P<0.05). Compared with group RIR, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly decreased in B, BE and BA groups, the expression of SIRT1 and Nrf2 in group B, Nrf2 and ASC in BE group and SIRT1, ASC and caspase-1 in BA group was up-regulated, and the expression of ASC, caspase-1 and NLRP3 in group B, SIRT1 and NLRP3 in BE group and Nrf2 in BA group was down-regulated ( P<0.05). Compared with group B, the serum concentrations of Cr and BUN, the contents of IL-1β and TNF-α and renal tubular injury score were significantly increased in BE and BA groups, the expression of ASC, caspase-1 and NLRP3 in BE and BA groups was up-regulated, and the expression of SIRT1 in BE and Nrf2 in BA groups was down-regulated ( P<0.05). Conclusion:SIRT1/Nrf2 signaling pathway is involved in the process of berberine-induced reduction of renal I/R, which is related to inhibiting pyroptosis in mice.
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Objective:To explore the evolution of imaging findings on dynamic chest high resolution CT（HRCT） in patients with mild COVID-19.Methods:The data of epidemiology, clinical data and continuous dynamic chest high-resolution CT images of the patients with mild COVID-19 were analyzed retrospectively. Twenty-five patients (including 24 common type and 1 mild) were enrolled in the group, including 14 males and 11 females, with age of （42±12） years and hospital stay of （19±5） days. The basic images and dynamic images of HRCT were analyzed and compared by the radiologists.Results:The clinical manifestations were fever (22 cases), cough (18 cases), expectoration (8 cases), pharyngeal pain (6 cases). Most laboratory tests lacked specificity. There were no significant abnormalities on chest CT of one mild patient. HRCT findings of the common type were as follows: (1) the distribution of the lesions: most of the multiple lesions involved both lungs (19 cases), with average of 3 (3±1) lobes, located in the peripheral pulmonary zone near the pleura (22 cases); (2) the morphology and density of the lesions: most of the lesions were ground glass density foci (22 cases), which were patchy and massive (18 cases), nodular (10 cases) and arc broadband (7 cases); with the development of the disease, the density of some lesions increased with localized pulmonary consolidation (10 cases), accompanied by air bronchus sign (5 cases) and halo sign (5 cases). Dynamic changes of HRCT images in the chest: the positive manifestations were found on the 5th (5, 6) day after the onset of the disease, the progressive time of CT lesions was 5 (5, 7) days, the peak time of CT lesions was 11 (10, 13) days, and the turning time of CT lesions was 9 (8, 11) days.Conclusions:Dynamic chest HRCT can monitor the basic evolution process of the disease in patients with mild COVID-19, and provide a more intuitive basis for clinical early diagnosis and treatment.
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Objective To report a newly developed method and procedure to establish a rat model of subarachnoid hemorrhage in detail, and to provide a better model simulating the clinical subarachnoid hemorrhage caused by a ruptured aneurysm for related research.Methods One hundred and twenty healthy SPF 2-3-month old male Sprague-Dawley rats were divided into 4 groups, 30 rats in each group.The three experimental groups were sacrificed at 6, 24 and 72 hours after modeling.Rat models of subarachnoid hemorrhage were established by inserting a fiber core in the internal carotid artery and piercing this artery.Successful establishment of the subarachnoid hemorrhage model was confirmed by observation of breathing, pupil, defecation, urination and inspection at autopsy dissection.The controllability and reproducibility of this model were verified by observation of clinical manifestation and explored by mortality analysis.Results Subarachnoid hemorrhage was successfully induced by fiber core piercing the internal carotid artery at the needed location.Conclusions This method of model preparation is stable and understandable.The operation is nimble, with a good reproducibility.This model can be successfully performed after a short time learning, well simulate the sudden hemorrhage caused by a ruptured aneurysm, and suitable for research on early brain injury and vasospasm after subarachnoid hemorrhage.
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AIM To research the antiasthmatic effects of Zuogui Pills,Yougui Pills and Yupingfeng Powder and their mechanism of action.METHODS Male Brown Norway rats were randomly divided into five groups.OVA was used to induce the model of chronic asthma.Five groups of asthmatic rats were given Zuogui Pills,Yougui Pills,Yupingfeng Powder by gavage once daily for 14 d,respectively.The normal and the model group were given normal saline.Penh value,serum Thl,Th2 inflammatory cytokines,and CROT and ACTH protein levels were determined.Pathological changes of airway remodeling were measured.RT-PCR method was used to measure the expression of CRH mRNA in hypothalamus,and TGF-β1 and Smad3 mRNA in lung tissues.RESULTS Penh values of Yupingfeng Powder group and Yougui Pills group were significantly lower than that of Zuogui Pills group.The Thl inflammatory cytokine of Yupingfeng Powder group was significantly higher than that of Zuogui Pills group,while the Th2 cytokine was significantly lower than that of Zuogui Pills group.In Yougui Pills group,only IL-5 was significantly lower than that of Zuogui Pills group.Serum CROT,ACTH protein and CRH mRNA of Yupingfeng Powder group were significantly higher than those in Zuogui Pills group.No significant difference was found between Yupingfeng Powder group and Zuogui Pills group with regard to TGF-β1 and Smad3 mRNA.Smad3 mRNA of Yougui Pills group was significantly higher than that of the model group.Airway remodeling was significantly reduced by Yupingfeng Powder and Zuogui Pills.The number of goblet cells in Yougui Pills group was significantly lower than that in Zuogui Pills group.CONCLUSION All the three formulaes possess significant antiasthma effects.Regulation effects of Yupingfeng Powder on Thl/Th2 balance,HPA and TGF-β1/Smad3 pathway are strong.Compared with Zuogui Pills,Yougui Pills possesses weaker TGF-β1/Smad3 regulative effects and stronger HPA regulative effects.
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Objective To investigate the effects of fasudil hydrochloride on learning and memory, and the autophagy in hippocampal CA1 neurons in subarachnoid hemorrhage (SAH) rats. Methods Fifty-four male Sprague-Dawley rats were randomly divided into sham group (n=18), SAH group (n=18) and drug group (n=18). Subarachnoid hemorrhage model was established with internal carotid artery punc-ture. The drug group was injected fasudil hydrochloride 10 mg/kg intraperitoneally after modeling per 24 hours, while the sham group and SAH group were injected the same volume of saline. They were tested with shuttle box test 6, 24 and 72 hours after intervention, then the hippocampal CA1 area was stained with HE and immunohistochemistry to observe the morphology of cells and the expression of Beclin-1 and microtubule-associated protein 1 light chain 3 II (LC3-II). Results Compared with the sham group, the frequence of avoidance de-creased in SAH group at each time point (P<0.05), while the avoidance reaction time increased (P<0.05);the survival of neurons in hippo-campal CA1 area decreased (P<0.05), and the expression of Beclin-1 and LC3-II increased (P<0.05). Compared with SAH group, the fre-quence of avoidance increased in the drug group at each time point (P<0.05), while the avoidance reaction time decreased (P<0.05);the sur-vival of neurons in hippocampal CA1 area increased (P<0.05) and the expression of Beclin-1 and LC3-II increased further (P<0.05). Con-clusion Fasudil hydrochloride can improve learning and memory ability and protect neurons from damage, which may associate with the ex-cess of autophagy activation in hippocampal CA1 areas in SAH rats.
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Objective@#To evaluate the effect of breast massage at different time in the early period on maternal lactation after cesarean section.@*Methods@#80 women delivered by cesarean section were randomly selected from maternity ward of a hospital in Shandong province during Jan. 2013 to Jan. 2015; which were divided into four groups, with 20 patients in each. Three groups received 3 times of breast massage every 24 hoursbeginning from 2, 12 and 24 h after cesarean section, respectively. The control group didn't receive any breast massage. The starting time and status of lactation were observed and recorded after cesarean section. 5 ml venous blood sample was drawn from each patient respectively at 2 h before cesarean, 6, 12, 24, 48 and 72 h after cesarean to test the level of serum prolactin. The lactation status of each group was compared.@*Results@#The P50 (P25-P75) of starting time of lactation of the three massage groups and control group were 3 (2-6) h, 4 (2-8) h, 4 (3-12) h and 4 (2-12) h, respectively, whose differences showed no statistical significance (H=3.32, P=0.345).The number of delivered women with adequate lactation 24 hours after cesarean was 10 in the group who received massage beginning from 2 h after cesarean; while the number was only 2 in the control group. The number of delivered women with adequate lactation 48 hours after cesarean was 18 in the group who received massage beginning from 2 h after cesarean; while the number was 8 in the control group. The differences showed statistical significances (P values were 0.021 and 0.008, respectively). The serum prolactin level in the group of delivered women who received massage from 2 h after cesarean was separately (195.9±78.5), (176.0±96.5), (216.4±110.0), (190.0±56.8) and (184.8±69.6) μg/L at 2, 12, 24, 48 and 72 h after cesarean, which were significantly higher than those in the control group (which were (128.8±40.6), (127.3±66.8), (162.2±58.8), (145.1±64.7) and (141.7±49.3) μg/L, respectively) (P=0.007).@*Conclusion@#Breast massage beginning from 2 hours after cesarean section can effectively improve the lactation status of delivered women.
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Objective To investigate effects of capsaicin on proliferation,migration and invasion of human large cell carcinoma NCI-H460 and discuss the possible mechanisms by which capsaicin inhibits non-small cell lung cancer. Methods NCI-H460 cells were cultured in vitro and treated with capsaicin at various concentrations. Colony formation assay,wound healing assay,and transwell chamber invasion assay were used to detect the effects of capsaicin on proliferation,migration and invasion of NCI-H460 cells. Western blotting was used to detect the protein expression of E-cadherin,N-cadherin,Vimentin and Snail. Results The rate of colony formation of 100,200,300 μmol.L-1 of capsaicin were (91.17±24.38)%,(43.22±12.91)% and (15.71±4.26)%,respectively,the control group was (99.53±21.25)%.the rate of migration of NCI-H460 cells of 25,50, 100 μmol.L-1 of capsaicin were (85.83±17.43)%,(37.92±10.16)% and(21.08±6.39)%,respectively,the control group was (93.04±20.23)%.The number of invasion of NCI-H460 cells of 25,50,100 μmol.L-1 of capsaicin were (99±18.2),(75± 17.9) and( 52 ± 14. 1 ) , respectively, the control group was ( 107 ± 20. 4 ) . The middle and high-dose capsaicincould inhibit proliferation,migration and invasion of NCI-H460 cells ( P<0.05) ,and E-cadherin expression level was significantly up-regulated and N-cadherin, Vimentin, Snail expression level was significantly down-regulated ( P<0. 05 ) . Conclusion Capsaicin can inhibit proliferation,migration and invasion of NCI-H460 cells,the mechanism may be related to up-regulation of E-cadherin and down-regulation of N-cadherin,Vimentin and Snail expression levels.
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Objective To explore the variation trends of interleukin-1β( IL-1β) and intercellular adhesion molecule-1 (ICAM-1) in both normal and affected sides of brain tissues in rats with ischemia-reperfusion injury and the therapeutic action of electroacupuncture.Methods The cerebral ischemia-reperfusion model was established with suture embolization in the right middle cerebral artery.The rats were randomly divided into control group, model group and electroacupunture group.Each group was then divided into six subgroups by the time after operation (12 h,24 h,48 h,72 h,96 h,144 h), ten rats in each subgroup. Frozen sections of brain tissues were prepared and the expression of IL-1βand ICAM-1 in brain tissues of both sides were detec-ted by immunohistochemistry.Results The expressions of IL-1βand ICAM-1 showed typical bimodal pattern in both affected is-chemic region and contralateral normal region.In the model group, the peaks of IL-1βin the cerebral ischemic region were at 12 h and 48 h, while in the contralateral normal region the peaks were at 12 h and 144 h, the expression of IL-1βin the ischemic region was significantly higher than that in the contralateral normal region at 48 h (P<0.05), and lower at 96 h and 144 h (P <0.05).In the electroacupuncture group, the expressions of IL-1βin the ipsilateral region were significantly lower than that in the contralateral region at 24 h, 48 h and 144 h (P<0.05).In the model group, the peaks of ICAM-1 in the cerebral ischemic regions were at 24 h and 72 h, while in the contralateral normal regions the peaks were at 24 h and 144 h.In the electroacupunc-ture group, the expressions of ICAM-1 in the ischemic regions were significantly lower than that in the contralateral normal re-gions at all the 12 h, 24 h, 48 h, 72 h and 144 h (P<0.05).Conclusions Our findings suggest that electroacupuncture may inhibit the inflammation of ischemia/reperfusion brain tissue through reducing the expression of IL-1βand ICAM-1 to relieve the cerebral ischemia-reperfusion injury.
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Most of palatal fistula occur from poor repairation of cleft palate, leaving an abnormal channel between mouth and nose. Palatal fistula can cause a series of complications, such as voice and hearing disorder, poor oral and nasal hygiene, psychological diseases and so on. However, hard palate fistula secondary to Leukemia infection is rarely seen, it hasn't been reported yet. We report one case with hard palate fistula and nasal septum perforation after Leukemia infection.
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Humans , Fistula , Infections , Leukemia , Nasal Septal Perforation , Nose , Nose Diseases , Oral Fistula , Palate, Hard , Pathology , Postoperative ComplicationsABSTRACT
ObjectiveTo investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure (FHF) rat, and to study the effect on liver function and hepatocyte proliferation. MethodsMesenchymal stem cells(MSCs)were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups: MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group (n=8 per group), blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group(P＜0. 05), but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group(P＜0.01), but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group (P=0. 049), but there was no significant difference between the MSC-CM and PHGF group. ConclusionsThe paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.
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Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.
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<p><b>BACKGROUND AND OBJECTIVE</b>It has been proven that cytochrome P450 enzyme 2A13 (CYP2A13) played an important role in the association between single nucleotide polymorphisms (SNP) and human diseases. Cytochrome P450 enzymes are a group of isoenzymes, whose sequence homology may interfere with the study for SNP. The aim of this study is to explore the interference on the SNP study of CYP2A13 caused by homologous sequences.</p><p><b>METHODS</b>Taqman probe was applied to detect distribution of rs8192789 sites in 573 subjects, and BLAST method was used to analyze the amplified sequences. Partial sequences of CYP2A13 were emplified by PCR from 60 cases. The emplified sequences were TA cloned and sequenced.</p><p><b>RESULTS</b>For rs8192789 loci in 573 cases, only 3 cases were TT, while the rest were CT heterozygotes, which was caused by homologous sequences. There are a large number of overlapping peaks in identical sequences of 60 cases, and the SNP of 101 amino acid site reported in the SNP database is not found. The cloned sequences are 247 bp, 235 bp fragments.</p><p><b>CONCLUSION</b>The homologous sequences may interfere the study for SNP of CYP2A13, and some SNP may not exist.</p>
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Humans , Aryl Hydrocarbon Hydroxylases , Genetics , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , GeneticsABSTRACT
Objective To investigate the safety and tolerability of darbepoetin alfa.a long-lasting erythmpoietin,single intravenous administration in maintenance hemodialysis(MHD) patients. Methods A single center,open clinical trial was carried out.Forty-three stable MHD patients were divided into 5 groups and received darbepoetin α at dosage of 0.225,0.45,0.9,1.8,3.6 μg/kg respectively.The vital signs,symptoms,ECG and laboratory examinations were monitored and detected before and after administration. Results Of the 43 patients (male 26and female 17),the largest tolerable dosage of darbepoetin alfa was 3.6 μg/kg.During the study,the main side effect associated with darbepoetin was hypertension aggravation(7%).One patient died but that was not associated with darbepoetin alfa. Conclusion Darbepoetin alfa is sale and well tolerated.
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Objective To study the effects of rehabilitation training on the expression of Nogo-A around the area of cerebral infraction using rats. Methods A total of 60 Wistar rats were randomly divided into a reha- bilitation group and a control group after an experimental cerebral infarction had been established in them. The ani- mals in the rehabilitation group were given exercise with a rotating bar, a balance beam and a rolling cage for one hour daily, while those in the control group were caged without any abnormal exercise. Nogo-A expression in the ar- ea surrounding the infarcts was detected by imunohistochemical techniques at the 3rd, 7th, 14th, 21st and 28th day after infarction. Meanwhile, neurobehavioral evaluations were also conducted. Results The animals in the rehabilitation group scored much lower than the controls in the behavioral evaluations at the 14th, 21st and 28th day. The expression of Nogo-A in tissues around the infracted area increased by the 7th day and peaked at the 21st day in both groups, but the expression of Nogo-A was significantly stronger in the rehabilitation group at the 14th, 21st and 28th days. Conclusion Rehabilitation training decreased the expression of Nogo-A in the brain of rats after infarction. This may have important implications for the functional recovery of the central nervous system.
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OBJECTIVE:To observe the clinical efficacy of Tianmeng capsule in the treatment of climacteric female with burning mouth syndrome(BMS).METHODS:A total of 248 climacteric female of patients with BMS were randomly assigned to receive Tianmeng capsule plus comprehensive psychotherapy(experiment group,n=124)or VitB1+Oryzanol +VitE plus comprehensive psychotherapy(control group,n=124)for 1 month.RESULTS:The total effective rate was 91.9% for the experiment group versus 48.4% for the control group(P
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AIM: To set up a new method to analyze IR fingerprint spectra, which was in line with the cha racteristic of traditional Chinese medicine. METHODS: In this paper. two new indexes, common peak ratio and variant peak ratio, were applied and its values were calculated by means of the sequential analysis, in which each sample's IR fingerprint spectra was set up and the common peak ratio sequences were arranged in order of size in comparison with other samples. RESULTS: The method could distinguish the water extract of Radix Glycyrrhizae of different varieties and different areas. CONCLUSION: The dual index sequential analysis enables us to distinct two or more herb's IR fingerprint.
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Objective To investigate the effects of lipopolysaccharide(LPS) on Src-suppressed C Kinase Substrate(SSeCKS) in cultured astrocytes. Methods Purified astrocytes were randomly divided into three groups:control group,singly stressed and multiple stressed groups.The expression of SSeCKS was detected by Real time RT-PCR and Western blotting,while immunocytochemistry was used to investigate the subcellular localization of it. Results Real time RT-PCR indicated that,after 12 hours incubation,both 100 ?g/L and 1mg/L LPS were able to elevate the levels of SSeCKS mRNA compared with control group,while 1mg/L LPS did not have a stronger effect than 100 ?g/L.Western blotting analysis showed 100 ?g/L LPS significantly increased the expression of SSeCKS.In time response experiments,the levels of SSeCKS expression enhanced three hours after the stimulation,peaked at the sixth hour,coincident with its rapid phosphorylation,and remained high until the 24th hour.Immunocytochemistry suggested a perinuclear translocation of SSeCKS,while the PKC inhibitor RO-31-8220 blocked it.Conclusion In cultured astrocytes,LPS can enhance the expression of SSeCKS,increase its phosphorylation level and change its subcellular localization.These effects are correlated with the PKC pathway,which indicates SSeCKS might participate in the signal transduction of inflammation in cultured astrocytes.