Objective To investigate the effect and mechanism of extract ginkgo biloba (EGb) on learning memory ability and hippocampus CA1 region Caspase-9P35 activity in dementia rats induced by D-gal.Methods 48 rats were randomly divided into six groups.The dementia model were established by injecting intraperitoneally with the D-gal and each EGb group was injected different doses of EGb simultaneously.TUNEL method was used to detect apoptosis of hippocampal neurons in the CA1 region.Immunohistochemistry and Western Blot were used to determine the expression of Caspase-9P35 in hippocampus CA1 region.Results Compared with the normal group,TUN EL-positive neurons ( (37.8 ± 1.3 ),(0.8 ± 0.2 ) ) and the activity of Caspase-9P35 ( (37.6 ±1.8 ),(6.2 ± 1.3 ) ) had significant increase in hippocampal CA1 subfield of D-gal group (P ＜ 0.01 ).Contrast to D-Gal group,TUNEL-positive neurons ( ( 17.6 ± 0.9),(9.8 ± 0.8 ),( 37.8 ± 1.3 ) ) and the activity of Caspase9P35( (28.6 ± 1.3),(25.0 ± 1.6),(37.6 ± 1.8) ) were significant decreased in EGb-M and EGb-H group (P＜0.01 ).While TUNEL-positive neurons and the activity of Caspase-9P35 had not significant difference in the therapy group than D-Gal group (P ＞ 0.05).Conclusion EGb can improve the cognitive level of the dementia rats.One of the therapeutic mechanisms may be to decrease the hippocampus CA1 region Caspase-9P35 activity.The results of the pretreatment group was more effective than the therapy group.

BACKGROUND:The correlation between blood stasis syndrome and non-blood stasis syndrome of lumbar intervertebral disc protrusion remains unclear.
OBJECTIVE:To construct serum protein pattern model for diagnosing blood stasis syndrome of lumbar intervertebral disc protrusion.
METHODS:A total of 180 cases were included in this study and divided into treatment group (120 patients with lumbar intervertebral disc protrusion) and control group (60 healthy cases from physical examination). Furthermore treatment group was equal y assigned into blood stasis syndrome subgroup and non-blood stasis syndrome subgroup, with 60 cases in each subgroup. The involved cases were wel matched in nations, genders and ages. Serum samples of peripheral blood from the 180 cases were col ected. Surface-enhanced laser desorption/inionation time of flight mass spectrometry and ProteinChip technology were employed to detect and plot protein mass spectrum. The protein peak values were identified using Biomarker Wizard software. Then serum diagnosis model of blood stasis syndrome of lumbar intervertebral disc protrusion was established. The obtained models were verified through double blind method. The differential proteins were searched by ExPASy data.
RESULTS AND CONCLUSION:We detected that peak values of eleven proteins had statistical significance (P<0.05) from the involved 180 cases. Among them, two proteins were highly expressed while the other nine proteins were lowly expressed. Serum protein pattern model for diagnosing blood stasis syndrome of lumbar intervertebral disc protrusion was established through Biomarker Patterns software, and the sensibility was 86.667%, the specificity was 94.167%, the positive predictive value was 88.136%. There are a variety of abnormal y expressed proteins in the serum of the patients with blood stasis syndrome of lumbar intervertebral disc protrusion. The serum protein pattern model involved eleven different proteins can be used to diagnose blood stasis syndrome of lumbar intervertebral disc protrusion.

OBJECTIVETo establish a real-time quantitative RT-PCR method for the detection of expression of bcl-2 mRNA.

METHODSThe vector containing bcl-2 gene as standard template was constructed with T-A cloning technique. The fluorogenic probe (i.e.,Taq man probe) was used to establish a real-time RT-PCR. bcl-2 mRNA expression level in Burkitt's lymphoma pre-and post-treated with bcl-2 antisense phosphothioate oligodeoxynucleotides (AS-PS-ODN) was determined with real-time quantitative RT-PCR, also with semi-quantitative RT-PCR.

RESULTSThe expression of bcl-2 mRNA in Burkitt's lymphoma treated with bcl-2 AS- PS-ODN decreased significantly and no changes of bcl-2-mRNA expression in group treated with nonsense ODN were noticed. The semi-quantitative RT-PCR results also demonstrated that bcl-2 level varied as detected with real-time fluorogenic quantitative RT-PCR, but less sensitive and accurate.

CONCLUSIONDetection of bcl-2 mRNA expression with the fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR.

Electrophoresis, Agar Gel ; methods ; Gene Expression ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

We constructed the eukaryotic expression vector of human IL-35-IgG4 (Fc)-pOptiVEC-TOPO by gene recombination technique and expressed the fusion protein human IL-35-IgG4 (Fc) in CHO/DG44 cells. The two components of the newly discovered cytokine human IL-35, EBI3 and IL-12p35, were amplified by PCR from the cDNA library derived from the KG-I cells after LPS induction. The two PCR-amplified cDNA fragments of human IL-35 were linked by over-lapping PCR and then cloned into the IgG4 (Fc)-pOptiVEC-TOPO vector. The constructed plasmid with the recombinant cDNA IL-35-IgG4 (Fc) was verified by restriction enzyme digestion analysis, PCR and DNA sequencing. The verified plasmid with the recombinant cDNA was transfected into CHO/DG44 cells using Lipofectamine 2000. The success of the transfection was examined and confirmed by RT-PCR. After selection in alpha-MEM (-) medium, the IL-35-Ig G4 (Fc) positive CHO/DG44 clones were chosen and the media from these positive clones were collected to be used to purify the fusion protein. The positive CHO/DG44 clones were further cultured in increasing concentrations of MTX and the expression levels of the fusion protein IL-35-Ig G4 (Fc) were repetitively induced by MTX-induced gene amplification. The IL-35-IgG4 (Fc) fusion protein was purified from the media collected from the positive CHO/DG44 clones by protein G affinity chromatography and then identified by SDS-PAGE and Western blotting. The results showed that one protein band was found to match well with the predicted relative molecular mass of human IL-35-IgG4 (Fc) and this protein could specifically bind to anti-human IgG4 (Fc) monoclonal antibody. In conclusion, our study successfully established an IL-35-IgG4 (Fc) positive DG44 cell line which could stably express IL-35-IgG4 (Fc) fusion protein.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Gene Fusion ; genetics ; Genetic Vectors ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Interleukins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Volume dynamics is a two-compartment dynamical model using hemoglobin (Hb) derived plasma diluted level as input data and urine output as input variable through consecutive repeated measurements of Hb concentration in the blood during infusion. It could be applied to evaluate and guide crystalloid fluid rehydration for patients with dehydration or hypovolemia and during anesthesia or surgery. Volume dynamics could be also used to quantificate of strains, hypovolume, and the change of fluid distribution and elimination caused by anesthesia or surgery. The factors which influence the volume resuscitation are complex, including gender, age, hemodynamic state [mean arterial pressure (MAP)], health and stress state, renal function, consciousness, surgical or anesthesia state and so on, which may affect the half-life, distribution, and volume of the fluid. This article summarizes and analyzes the pathophysiological changes of crystalloids fluid in vivo, in order to provide reference for volume management in critically ill patients.

Hypersplenism is an important complication of cirrhotic portal hypertension, and splenectomy is an important means to treat hypersplenism in cirrhosis. It is realized that hypersplenism played a pathological role in the course of cirrhosis. This article analyzes and compares the changes in the condition of patients with cirrhosis between splenectomy with and without hyperfunction, and comprehensively discusses the pathological role and mechanism of hypersplenism in the course of cirrhosis, in order to strengthen the clinical prevention and treatment of hypersplenism in cirrhosis and to better improve the condition and prognosis of patients with cirrhosis.

Objective:To detect serum levels of vitamin A (Vit A), vitamin D(Vit D)25-hydroxy vitamin D[25-(OH)D] and vitamin E(Vit E) in children aged 0-6 years in Tibetan Plateau of Garzi Prefecture, thus providing references for physical examinations and prevention of 4 key diseases (rickets, malnutrition anemia, pneumonia and diarrhea) in children in plateau areas by relevant government departments.Methods:A total of 2 122 children who participated in physical examination in 12 townships of Xiangcheng County and 14 townships of Daocheng County, Garzi Tibetan Autonomous Prefecture, Sichuan Province from April 2017 to April 2019 with 0-6 years old were recruited for surveying physical measurements and collection of venous blood.Serum Vit A and Vit E levels were detected by high performance liquid chromatography.Serum levels of 25-(OH)D were detected by high performance liquid chromatography tandem mass spectrometry.The relationship between Vit A, Vit E and 25-(OH)D levels with the gender, age, seasonal change and altitude was analyzed.Results:The serum Vit A level, subclinical Vit A deficiency rate and marginal vitamin A deficiency rate were(1.05±0.27) μmol/L, 8.15%(173/2 122 cases) and 45.99%(976/2 122 cases), respectively in 2 122 children with 0-6 years old.There were significant differences in the serum Vit A level, the subclinical Vit A deficiency rate and the marginal vitamin A deficiency rate in children with different ages, seasons and altitudes (all P<0.05). The serum level of 25-(OH)D and 25-(OH)D deficiency rate insufficient rate were (24.65±6.45) ng/L, 6.03%(128/2 122 cases) and 16.59%(352/2 122 cases), respectively.There were significant differences in the serum level of 25-(OH)D, 25-(OH)D deficiency rate and 25-(OH)D insufficient rate in children with different ages and seasons (all P<0.05). The mean serum Vit E level, Vit E deficiency rate and Vit E insufficient rate were (7.81±1.74) mg/L, 2.78%(59/2 122 cases) and 29.59%(628/2 122 cases), respectively.There were significant differences in serum Vit E level, Vit E deficiency rate and Vit E insufficient rate in children with different ages and seasons (all P<0.05). The mean serum levels of Vit A and Vit D remained the lowest before the age of 1 year, and their deficiencies at this age were the most significant.The mean serum level of Vit E remained the lowest in >1-2 years old, and its deficiency and insufficient at this age were the most significant.Vit A, D and E levels were significantly affected by seasonal changes, which were significantly higher in the summer than in the spring, autumn and winter.In addition, Vit A and 25-(OH)D were significantly affected by the altitude, which were the lowest above 4 km altitude. Conclusions:The overall serum levels of Vit A, 25-(OH) D and E in children with 0-6 years old in Tibetan Plateau areas of Ganzi Prefecture are lower than those in plain areas.Vit A, 25-(OH) D and Vit E levels significantly differed in the age, season and altitude, which are related to the lack of local resources, insufficient maternal nutrition during pregnancy and insufficient intake after birth, as well as temperature and light caused by changes in local seasons and altitude.Therefore, it is necessary to make reasonable supplements during pregnancy to prevent vitamin deficiency.