Objective To improve the quality standard for quality control of Baji Bushen Pills.Methods Radix Rehmanniae Praeparata and Radix Angelicae Sinensis in this medicine were identified by TLC,the 2,3,5,4’-Tetrahydroxystilbene-2-O-?-D-glucoside content in this medicine was determined by HPLC.The acetonitril-water(18:82)solution was used as the mobile phase,the column was C18,the flow rate was 1.0 mL/min,and the detective wavelength was at 320 nm.Results Radix Rehmanniae Praeparata and Radix Angelicae Sinensis could be identified by TLC.2,3,5,4’-Tetrahydroxystilbene-2-O-?-D-glucoside showed a good linear relationship at the range of 11.28 ?g/mL～282.00 ?g/mL(r=0.999 9).The average recovery was 96.1 %with RSD of 0.8 %(n=6).Conclusion The method is simple,reliable,accurate and can be applied to the quality control of the preparation.

BACKGROUND: The abnormal apoptosis of intestinal epithelial cells is the main cause of intestinal mucous membrane injury during ischemia reperfusion. Shenfu injection has good therapeutic effect on intestinal mucous membrane injury. OBJECTIVE: To investigate the influence of shenfu injection medication on intestinal apoptotic epithelial number, apoptosis-related caspase-3 and Bcl-2 gene expression, as well as the level of tumor necrosis factor (TNF)in intestinal ischemia reperfusion rat model.DESIGN: Randomized controlled experiment.SETTING:Department of Anesthesia, Renmin Hospital of Wuhan University.MATERIALS:This experiment was carried out in the laboratory of the Department of Anesthesia, Renmin Hospital of Wuhan University, between December 2002 and June 2003. Totally 24 healthy SD rats of clean grade were randomized into blank control group, intestinal ischemia reperfusion (IR) group and shenfu injection group with 8 rats in each group.METHODS: Rats in each group were anesthetized with intraperitoneal injection of ethyl carbamate at dosage of 1mg/kg; then in intestinal IR group and shenfu injection group rats' superior mesenteric artery was occluded with vascular clamp for 1 hour before 2-hour reperfusion,which was not conducted in blank control group. Rats in shenfu injection group were intravenously injected with shenfu injection at 30 minutes before occlusion at dose of 0.02 mL/g, which was replaced by the same volume of physical saline in blank control group and intestinal IR group. The expression of caspase-3 and Bcl-2 protein and cell apoptosis wee detected.MAIN OUTCOME MEASURES: ① Comparison of apoptotic index between groups. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium. ③ Comparison of TNF content in intestinal mucous membrane homogenate between groups.RESULTS: Totally 24 rats were included in this experiment and all ehtered the result analysis. ①Comparison of apoptotic index between groups:The apoptotic index was obviously lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(7.75-±1.89)%,(28.25±8.50)%, (4.25-±2.63)%, P ＜ 0.01]. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium: caspase-3 expression was lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(0.211 6±0.087 5), (0.354 7±0.077 8), (0.194 1±0.057 4) A,P ＜ 0.01, P ＞ 0.05]; Bcl-2 expression was remarkably higher in intestinal IR group than in blank control group (P ＜ 0.05), but obviously reduced in shenfu injection group compared to intestinal IR group (P ＜ 0.01). ③ TNF content of intestinal mucous membranehomogenate in each group: TNF content was remarkably higher in intestinal IR group than in blank control group and shenfu injection group [(189.7±56.3), (38.6±10.4), (47.5±l8.7)mg/L,P ＜ 0.01], and basically the same in shenfu inj~tion group and blank control group (P ＞ 0.05).CONCLUSION: Shenfu injection can suppress intestinal epithelial apoptosis by reducing TNF content and caspase-3 e.pression as well as upregulating the expression of Bcl-2 gene during ischemia reperfusion, thereby attenuating ischemia reperfusion injury of intestinal mucous membrane.

Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

Objective To observe the expression changes in microRNA (miR)-379 in the developmental process of the mouse heart and during the differentiation of P19 cells into cardiac myocytes,and to explore the possible relationship between miR-379 and the differentiation of cardiacmyocytes.Methods Heart tissues were collected from fetal mice in pregnant ones at their gestational age (8.5,11.5,14.5 and 18.5 days) respectively.Heart tissue sections of the fatal mice were obtained to observe the heart development process.Then total RNA was isolated from heart tissues by using the TRIzol method.Complementary DNA was synthesized from 1 μg total RNA by using a Reverse Transcriptase Kit.Finally,real-time PCR (RT-PCR) was employed to detect the expression of miR-379.At the same time,P19 cells were cultured with 10 mL/L Dimethyl sulfoxide in suspension for 4 days to form cell aggregation,and these aggregations were transferred into 6-wells plate for culturing by adherence.Beating cells were detected with microscopy on the 10th day after induction.Afterwards,total RNA was extracted from cultured P19 cells at different time points.Reverse transcription was executed to get DNA.At last,RT-PCR was used to explore the expression of miR-379 on 0,4,6,10 days after aggregation.Results The expression level of miR-379 was down-regulated gradually in the developing heart (at gestational age of 8.5,11.5,14.5,16.5 days,respectively),and there were significant differences on the different days (F =21.13,P ＜ 0.05).On the other hand,myocardial markers of troponin T represented an increasing trend during the process of P19 cells induction,which demonstrated that P19 cells were successfully induced into cardiomyocyte-like cells.Meanwhile,miR-379 showed a low expression on day 0 of P19 cells aggregation.On day 4,miR-379 demonstrated a higher level.Afterwards,miR-379 proved to be down-regulated gradually.Conclusions miR-379 plays a role in the process of the heart development,but the specific mechanisms need further research.

Objective To study the effects and security of umbilical cord mesenchymal stem cells transPlantation (UC-MSCs) for Patients with refractory systemic luPus erythematosus(SLE). Methods Forty Patients with refractory SLE were divided into two grouPs at random.All of Patients were treated for glucocorticoid and CTX, then Patients of the 2 grouP were transPlanted for UC-MSCs.All of Patients were observed before and 2 weeks after treatment,1 month,2 months,3 months,6 months,9 months and 12 months of clinical manifestations and laboratory Parameters . Results 2 weeks after treatment, 60%of the 1 grouP were SLEDAI<10 scores, and the 2 grouPs were 90%. There was statistically significant difference between the two grouPs (χ2=7.619, P=0.006). 1 year after treatment, PLT,ALB and C3 of the 2 grouPs were more than the 1 grouPs(P<0.05). SLEDAI,U-Pro,ESR and Hs-CRP of the 1 grouPs were more than the 2 grouPs(P<0.05). WBC,Cr and C4 of the two grouPs had not statistically significant difference(P﹥0.05). The recurrence rate of the 1 grouPs was 45%. Of the 2 grouPs was 15%.There was statistically significant difference between the two grouPs (χ2=4.286, P=0.038). Conclusion It is effective and safe for refractory SLE to transPlant UC-MSCs after using glucocorticoid and CTX. Further observation is required to evaluate long term efficacy and adverse reaction of UC-MSCs.

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.

Memory function is the core component of human brain cognition.Memory impairment significantly affects people's daily life and social ability,but existing cognitive training or drug therapy can not effectively improve memory function.In recent years,new techniques and methods have been emerging in brain science research,especially neural regulation techniques,such as transcranial electrical / magnetic stimulation,which can significantly regulate cognitive ability and effectively promote memory function.Transcranial direct current stimulation is a kind of noninvasive and safe technique.It can regulate cerebral cortex activity by changing the cerebral cortex excitability,and play a role in regulating brain function.At present,there have been reports at home and abroad that transcranial direct current stimulation can promote memory function and improve memory effectively.This technique is applied to the diagnosis and treatment of clinical diseases,but its mechanism is not clear.This article reviews the mechanism and application of transcranial direct current stimulation (TDCS) in promoting different types of memory function in order to provide evidence for early intervention and treatment of memory impairment.

Essential palatal tremor is relatively rare in clinical practice, which manifests involuntary and rhythmic contraction of soft-palate along with auditory click. The cause is unknown and there is no specific treatment at present. This article reports a female patient with essential palatine tremor, who presented with involuntarily beating of soft palate, disappeared during sleep, had sensory tricks, and gradually developed mental and psychological problems such as anxiety disorders. After treatment with integrated traditional Chinese and Western medicine, the symptoms improved. The clinical features of the case were analyzed, relevant literature was reviewed, and the possible etiology and characteristics of the disease were explored, so as to provide reference for clinical diagnosis and treatment.

Objective:To investigate the correlation between triglyceride-glucose (TyG) index and high on-treatment platelet reactivity (HTPR) during clopidogrel treatment in patients with ischemic stroke.Methods:Patients with ischemic stroke who received maintenance dose of clopidogrel (75 mg/d) in the Department of Neurology, Guangdong Provincial Hospital of Chinese Medicine from January 2017 to March 2021 were retrospectively included. The highest quartile (Q4) of the TyG index was defined as insulin resistance. Platelet reactivity was assessed by thromboelastogram and clopidogrel HTPR was defined as the clot strength induced by adenosine diphosphate (MA ADP) >47 mm. Multivariate regression model was used to analyze the independent correlation between TyG index and platelet reactivity. Results:A total of 83 patients were included. The TyG index showed a linear correlation with MA ADP. The patients were divided into 4 groups according to the quartile of TyG index. The incidence of clopidogrel HTPR increased significantly with the increase of the quartile of the TyG index ( Ptrend=0.017). Multivariate analysis showed that there was a significant independent correlation between insulin resistance and clopidogrel HTPR (odds ratio 4.597, 95% confidence interval 1.285-16.446; P=0.019). Conclusions:In patients with ischemic stroke treated with clopidogrel, the incidence of clopidogrel HTPR gradually increases with the increase of the quartile of the TyG index. The insulin resistance assessed by the TyG index is independently associated with clopidogrel HTPR.