Objective To study the effect of adenoviral-delivered short hairpin RNA (shRNA) target against permeability glycoprotein (Pgp) as a new drug in anti-epileptic drug resistance epilepsy treatment and to evaluate its efficiency. Methods MDR Sprague-Dawley (SD) rat estrocyte model was induced by Coriaria Lactone (CL), mainly over-expressing mdrlb. To reverse the drug resistance, astrecytes were treated with constructed replication deficient adencvirus AdS-EGFP-shRNAI-U6 delivering short hairpin (shRNA) target agianst mdrlb gene. Total RNA and protein were extracted from the infected cells, mdr1 b level was detected by Quantitative Real-time PCR whereas Pgp by Western blot, Rhodamine123 (Rho123) efflux ratio by Flow Cytometry. Results AdS-EGFP-shRNA1-U6 was succesfully constucted with high virus titer of 6×1010 pfu/ml. The interference efficency of AdS-EGFP-shRNA1-U6 agianst mdrlb in rat astrecyte model was about 94%. The Rho123 efllux ratio was about 15. 8%, significiently lower than control group which was 56. 2% (F = 127.5, P < 0. 05). Conclusions Pgp over-expression has been successfully suppressed and MDR has been reversed, which may provide a premising approach for refractory epilepsy remedy.

Objective To quantitatively study right ventricular function in the normal second and third trimester fetuses by M-mode tricuspid annular displacement(TAD).Methods TAD was measured using conventional M-mode echocardiography on 161 normal second and third trimester fetuses with gestation age (GA) between 19-38 weeks,meanwhile multiple parameters for evaluating right ventricular function were obtained using pulsed Doppler echocardiography (PW) and myocardial Dopple tissue imaging (DTI).The correlation between TAD and other parameters were analyzed using SPSS 17.0.Results In normal second and third trimester fetuses,the TAD was (9.38 ± 1.71)mm with a range of 5.79-13.90 mm,and was increased with the growth of GA.TAD was correlated with GA,E,A,Em,Am and Sm significantly (r =0.759,0.547,0.320,0.497,0.483 and 0.598 respectively,all P ＜0.001).TAD was not correlated with HR(P ＞0.05).TAD showed differences between the second trimester fetuses and the third trimester fetuses (P ＜ 0.05).Conclusions In normal second and third trimester fetuses,the TAD is increased with the growth of GA,and has good correlation with GA,E,A,Em,Am and Sm respectively,and may become a new promising modality to evaluate function of RV simply and accurately.The technique will be propitious to use in hospitals (without DTI) because of simplicity of operator and lower requirement on the technology and equipment precision.

AIM:To observe the effect of rapamycin (Rapa) on human neuroblastoma SH-SY5Y cell injury induced by oxygen-glucose deprivation (OGD), and to explore the role of autophagy in this process .METHODS: The SH-SY5Y cells were randomly divided into 4 groups:normal control group:the cells were cultured without OGD treatment;Rapa group:the cells were pretreated with Rapa for 1 h;OGD group:the culture medium was replaced by glucose-free me-dium and the cells were transferred to a humidified incubation chamber flushed by a gas mixture of 1%O2 , 94%N2 and 5% CO2 for 12 h; Rapa+OGD group: the cultured cells were treated with Rapa for 1 h, and then were given the same treatments as those in OGD group .The cell viability was assessed by MTT assay .The degree of the cell damage was evalu-ated by determining the leakage of lactate dehydrogenase ( LDH) .The enzyme activity of caspase-3 was detected .TUNEL staining were used to detect the variation of cell apoptosis .The protein levels of apoptosis-related proteins Bax and Bcl-2, autophagy-related protein beclin-1 and autophagy marker protein LC 3B were determined by Western blot .RESULTS:Compared with OGD group, the viability of the SH-SY5Y cells was significantly increased, and the activity of caspase-3 was significantly reduced in Rapa +OGD group (P<0.05).The SH-SY5Y cell injury was apparent after OGD with a great in-crease in the apoptotic rate (P<0.05).Compared with OGD group, the apoptotic rate significantly decreased in Rapa +OGD group (P<0.05).Compared with control group, the protein level of Bcl-2 was significantly decreased (P<0.05) and the protein level of Bax was significantly increased in OGD group .Compared with OGD group , the levels of Bcl-2, be-clin-1 and LC3B-Ⅱwere significantly increased and the protein level of Bax was significantly increased in Rapa +OGD group (P<0.05).CONCLUSION: Rapamycin has a protective effect on in vitro cultured SH-SY5Y cells injured by OGD.The mechanism may be related to the promotion of autophagy .

Objective To investigate the health-related quality of life and its relative factors in children with leukemia during maintenance treatment period. Therefore, the basis for effective individualization intervention can be provided. Methods A total of 224 leukemia children in maintenance treatment were collected in Affiliated Children′s Hospital of Chongqing Medical University from October 2015 to January 2016 by convenience sampling methods, questionnaire was used to assess the physical status, emotional status, social status, school status of the children. Results Single factor and generalized linear regression analysis were used to conclude that patient′s age (F=5.841-36.343, all P<0.01), parenting pattern (F=4.006-4.288, all P<0.05), family economic status (F=3.277-15.865, P<0.05 or 0.01) and the caregivers′information mastery level household location (F=2.044-2.661, P<0.05 or 0.01) had significant influence on the quality of life of children with leukemia in maintenance treatment. Conclusions The health-related quality of life and its relative factors in children with leukemia during maintenance treatment period should be analysis comprehensively. At the same time, take intervention should be taken from physiology, psychology, society, and school, to improve the social adaptability of children with leukemia and help them return to society as soon as possible.

BACKGROUND:Transpedicular transdiscal lumbar screw is a new type of spinal minimally invasive internal fixation technology.Compared with traditional bilateral pedicle screws,only one screw is needed to fix one segment on one side.It has the characteristics of being more economical,less trauma and easy to operate.However,studies on the application of transpedicular transdiscal lumbar screws combined with transforaminal lumbar interbody fusion(TLIF)and fixation are still rare. OBJECTIVE:To evaluate the effect of TLIF combined with various surgery methods on stress distribution of cage,fixation,disc lower and endplate and range of motion of lumbar vertebrae by constructing three kinds of finite element models including modified TLIF(cage alone)model,modified TLIF combined with bilateral pedicle screw(cage+BPS)model and modified TLIF combined with bilateral transpedicular transdiscal lumbar screw(cage+BTPTDS)model. METHODS:The CT images of the adult lumbar spine were used to establish the three kinds of TLIF finite element models:cage alone,cage+BPS and cage+BTPTDS using software Mimics,Geomagic and SolidWorks.ANSYS Workbench was used to simulate the application of six different motion loads of human body flexion and extension,left and right bending,and left and right rotation to calculate stress distribution and the changes in the range of motion of the lumbar spine of the cage,fixation,endplate and disc of the three lumbar spine surgery models and to compare the effects of three surgical options on the biomechanical effects of the lumbar spine. RESULTS AND CONCLUSION:(1)The cage alone model,cage+BPS model and cage+BTPTDS model were constructed successfully.(2)In flexion and lateral bending conditions,the maximum stress of the cage of cage+BTPTDS model was smaller than that of the cage alone model and a little greater than that of the cage+BPS model.In the extension condition,the maximum stress of the cage of the cage+BPS model was obviously smaller than that of the other two models.When it came to rotating condition,the maximum stress of the cage in the cage+BPS model and the cage+BTPTDS model presented no obvious difference,which was both smaller than the cage alone model.(3)The maximum stress of fixation of the cage+BTPTDS model was obviously bigger than the cage+BPS model in flexion and extension conditions,close to the cage+BPS model in lateral bending conditions,and smaller than the cage+BPS model in rotation conditions.(4)The maximum stress of the lower endplate of the fusion segment of the cage+BPS model was between the two other models.(5)In terms of the range of motion,the cage+BTPTDS model presented no obvious difference with that of the cage+BPS model at flexion and extension,left and right bending,and left and right rotation.(6)It is concluded that modified TLIF combined with transpedicular transdiscal lumbar screw provides stable support for the vertebral body of the fusion segment,ensures the motion range of the lumbar spine and has a good biomechanical effect.

OBJECTIVETo study the morphological variations of Paris polyphylla var. yunnansensis in different population for genetic diversity and breeding.

METHODThe characters of roots, stalks, leave and flowers were observed. The results were analyzed by DPS software.

RESULT AND CONCLUSIONP. polyphylla var. yunnansensis showed plenty genetic diversity, there existed obvious differences in morphological characters of different population. Principal components analysis showed that the number of calyces, petal, carpels, stamens is main factor,which causes the morphological variations in different population. Cluster analysis shows that 26 populations are incorporates in two types as 45.08 Euclidean distance. Leaf area index is distinct different in this two types.

Identification of the function of all genes in the mammalian genome is critical in understanding basic mechanisms of biology. However, the diploidy of mammalian somatic cells has greatly hindered efforts to elucidate the gene function in numerous biological processes by mutagenesis-based genetic approaches. Recently, mouse haploid embryonic stem (haES) cells have been successfully isolated from parthenogenetic and androgenetic embryos, providing an ideal tool for genetic analyses. In these studies, mouse haES cells have already shown that they could be used in cell-based forward or reverse genetic screenings and in generating gene-targeting via homologous recombination. In particular, haES cells from androgenetic embryos can be employed as novel, renewable form of fertilization agent for yielding live-born mice via injection into oocytes, thus showing the possibility that genetic analysis can be extended from cellular level to organism level.
Animals ; Embryonic Stem Cells ; cytology ; metabolism ; Genetic Techniques ; Genome ; Haploidy ; Models, Animal ; Mutagenesis ; Parthenogenesis

OBJECTIVE: To develop a fast, sensitive and cost-effective method based on resonance light scattering (RLS) for characterization of protein solubility to facilitate detection of changes in solubility of mutant proteins. METHODS: We examined the response curve of RLS intensities to the protein concentrations in synchronous scanning mode. The curve intersection points were searched to predict the maximal concentrations of the protein in dispersion state, which defined the solubility of the protein in this given state. Bovine serum albumin (BSA, 0-50 g/L) was used as the model to investigate the influences of pH values (6.5, 7.0, and 7.4) and salt concentrations (0.05, 0.10, 0.15, and 0.20 mol/L) on the determined solubility. The solubility of glutathione S-transferase isoenzymes alpha (GSTA, 0-27.0 g/L) and Mμ (GSTM, 0-20.0 g/L) were estimated for comparison. The RLS-based method was used to determine the solubility of uricase (MGU, 0-0.4 g/L) to provide assistance in improving the solubility of its mutants. RESULTS: We identified two intersection points in the RLS response curves of the tested proteins, among which the lower one represented an approximation of the maximal concentration (or the solubility of the protein) in single molecular dispersion, and the higher one the saturated concentration of the protein in multiple molecular aggregation. In HEPES buffer, the two intersection points of BSA (isoelectric point 4.6) both increased with the increase of pH (6.5-7.4), and their values were ~1.2 g/L and ~33 g/L at pH 7.4, respectively; the latter concentration approached the solubility of commercial BSA in the same buffer at the same pH. The addition of NaCl reduced the values of the two intersection points, and increasing salt ion concentration decreased the values of the lower intersection points. Further characterizations of GSTA and GSTM showed that the low concentration intersection points of the two proteins were ~0.7 g/L and ~0.8 g/L, and their high concentration intersection points were ~10 g/L and ~11 g/L, respectively, both lower than those of BSA, indicating the feasibility of the direct characterization of protein solubility by RLS. The two concentration intersection points of MGU were 0.24 g/L and 0.30 g/L, respectively, and the low concentration intersection point of its selected mutant was increased by 2 times. CONCLUSIONS RLS allows direct characterization of the solubility of macromolecular proteins. This method, which is simple and sensitive and needs only a small amount of proteins, has a unique advantage for rapid comparison of solubility of low-abundance protein mutants.
Hydrogen-Ion Concentration ; Light ; Scattering, Radiation ; Solubility ; Spectrum Analysis

OBJECTIVE: To investigate the effect of acetylcorynoline (Ace) for promoting functional recovery of injured spinal cord in rats and explore the underlying mechanism. METHODS: Rat models of spinal cord injury (SCI) were treated with intraperitoneal injection of different concentrations of Ace, with the sham-operated rats as the control group. After the treatment, the changes in motor function of the rats and the area of spinal cord injury were assessed with BBB score and HE staining, and the changes in pro-inflammatory cytokine levels and microglial activation were determined using PCR, ELISA and immunofluorescence staining. In a lipopolysaccharide (LPS)-treated BV2 cell model, the effects of different concentrations of Ace or DMSO on microglial activation and inflammatory cytokine production were observed. Network pharmacology analysis was performed to predict the target protein and signaling mechanism that mediated the inhibitory effect of Ace on microglia activation, and AutoDock software was used for molecular docking between Ace and the target protein. A signaling pathway blocker (Osimertinib) was used to verify the signaling mechanism in rat models of SCI and LPS-treated BV2 cell model. RESULTS: In rat models of SCI, Ace treatment significantly increased the BBB score, reduced the area of spinal cord injury, and lowered the number of activated microglia cells and the levels of pro-inflammatory cytokines (P < 0.05). The cell experiments showed that Ace treatment significantly lower the level of cell activation and the production of inflammatory cytokines in LPS-treated BV2 cells (P < 0.05). Network pharmacology analysis suggested that EGFR was the main target of Ace, and they bound to each other via hydrogen bonds as shown by molecular docking. Western blotting confirmed that Ace inhibited the activation of the EGFR/MAPK signaling pathway in injured mouse spinal cord tissue and in LPS-treated BV2 cells, and its inhibitory effect was comparable to that of Osimertinib. CONCLUSION In rat models of SCI, treatment with Ace can inhibit microglia-mediated inflammatory response by regulating the EGFR/MAPK pathway, thus promoting tissue repair and motor function recovery.
Mice ; Animals ; Rats ; Recovery of Function ; Lipopolysaccharides ; Microglia ; Molecular Docking Simulation ; Spinal Cord Injuries ; Signal Transduction ; Cytokines ; ErbB Receptors

OBJECTIVETo dynamically observe the effects of electroacupuncture (EA) on repair of gastric mucosal lesion in rats with gastric ulcer, and to explore the time-effect relationship and molecular mechanism of EA for gastric ulcer.

METHODSA total of 72 SD rats were randomly assigned to a normal group, a model group, a acupoint group and a sham acupoint group, and each group were further divided into a 1-day subgroup, a 4-day subgroup and a 7-day subgroup, 6 rats in each subgroup. The rat model of gastric ulcer was established by using intragastric administration of ethyl alcohol. The rats in the acupoint group were treated with EA at"Zusanli"(ST 36) and"Liangmen"(ST 21); the rats in the sham acupoint group were treated with EA at points 5 mm next to"Zusanli"(ST 36) and"Liangmen"(ST 21); the EA was given 30 min per treatment, once a day. The rats in the normal group and model group were treated with immobilization for 30 min per day, and no EA was given. PR-PCR method was applied to test the expression of proliferating cell nuclear antigen (PCNA) and substance P (SP); Western blot method was applied to test the expression of neurotensin (NT).

RESULTSAfter 1-day treatment, the ulcer index in the model group was higher than that in the normal group (<0.01), and the expression of PCNA, SP and NT was decreased (<0.01, <0.05); compared with the model group and sham acupoint group, the ulcer index was decreased in the acupoint group (both <0.05), and the expression of PCNA and SP was up-regulated (all <0.05) while that of NT was up-regulated (both <0.01). After 4-day treatment, the ulcer index in the model group was reduced but still higher than that in the normal group (<0.05), and the expression of PCNA, SP and NT was up-regulated and higher than that in the normal group (all <0.01); the ulcer index in the acupoint group was similar to that in the normal group (>0.05), and the expression of PCNA and SP was lower than that in the model group (<0.01, <0.05), and the expression of NT was not significantly different from that in the model group (>0.05). After 7-day treatment, the differences of indexes above were not significant among the four groups (all >0.05).

CONCLUSIONEA at acupoints of stomach meridian has two-way regulation on PCNA and SP and improve the expression of NT in different pathological state of gastric ulcer, which could further improve the repair of gastric ulcer.