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Lumbar spinal stenosis is one of the most common causes of low back and leg pain. However, there is a lack of evidence-based guidelines for the treatment of lumbar spinal stenosis with integrated traditional Chinese medicine and western medicine. To standardize the diagnosis and treatment of lumbar spinal stenosis with a combination of traditional Chinese medicine and western medicine, this guideline strictly followed the relevant guiding principles for the development of guidelines both domestically and internationally. A multidisciplinary working group was established. We adopted the GRADE approach to grade the quality of evidence and the strength of recommendation. Finally recommendations on the treatment of lumbar spinal stenosis with integrated traditional Chinese medicine and western medicine were formed, aiming to standardize clinical practice. This guideline is for patients with all degrees of lumbar spinal stenosis.

Objective The resurrections of ancient landslide deposits are one of the primary geological hazards in the Qinghai-Tibet Plateau and surrounding areas of China and pose significant safety threats to major transportation and water conservancy projects under construction in western China. Therefore, it is crucial to investigate the formation and evolution mechanism of ancient landslides and evaluate the stability of their deposits. This research can provide theoretical support for the early recognition and prevention of the resurrection of ancient landslide deposits. Methods The ancient landslide deposits in Jiangdingya, Zhouqu County, Gansu Province, have experienced local resurrection several times in the past decade, creating severe threats to the lives and property of local people by blocking the Bailong River. To determine the morphology and structural characteristics of the ancient Jiangdingya landslide deposits, this study utilized field investigations and unmanned aerial vehicle (UAV) tilt photography. Based on this, the evolution mechanism and dynamic process of the landslide were analysed, and the stability of the deposits was qualitatively evaluated using InSAR deformation data. Results The results show that the ancient landslide at Jiangdingya is a typical large-scale earthquake landslide, with its sliding body located in a downslope position in three directions, forming a multilevel stepped deposit shape. The dynamic process of an ancient landslide under seismic loads can be divided into several stages, including vibration and cracking in the upper-middle part, shearing and landslide initiation in the front edge locking segment, tearing and landslide acceleration in the rear edge, obstruction and landslide deceleration in the front edge, and stabilization. Conclusion Due to the overall downwards movement of the ancient landslide under seismic loads, there are a large number of intact rock masses in the upper deposits, which are relatively stable. However, the middle and lower deposits are mostly composed of weak structures such as fault fracture zones and fragmented rock masses, which have poor stability and are highly likely to resurrect in the future. This study provides important insights into the formation and evolution of ancient landslides and the evaluation of their stability, which can help prevent future landslides and protect local communities.

Objective To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. Methods Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. Results The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. Conclusion This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.

Objective To explore the role of CD200 in Ewing's sarcoma and its possible pathological mechanisms, and to search for potential drugs that can improve the survival prognosis of Ewing's sarcoma patients. Methods The RNA seq and clinical data of Ewing's sarcoma patients were obtained from the International Cancer Genome Consortium database. The survival analysis was used to explore the impact of CD200 on the survival prognosis of Ewing's sarcoma patients. The gene set enrichment analysis (GSEA) and Lincx analysis were carried out to find the molecular mechanism of CD200 promoting the malignant process of Ewing's sarcoma and the anti-Ewing's sarcoma drugs that may target CD200. The expression level of CD200 was verified by RT qPCR. Results CD200 was significantly overexpressed in Ewing's sarcoma cells (P

Objective To observe the effect of using the therapeutic apparatus for music therapy based on Traditional Chinese Medicine (TCM)syndrome differentiation in the treatment of functional abdominal distension. Methods Totally 60 patients with functional abdominal distension were randomly divided into the control group and the intervention group, with 30 cases in each group. All patients received a routine treatment and nursing (7 days per course, two courses), and the music therapy based on TCM syndrome differentiation by using a therapeutic apparatus was performed in the intervention group. The Gastrointestinal Symptom Assessment score and overall effective rate were compared between two groups. Results The Gastrointestinal Symptom Assessment score decreased in both two group after two courses of treatment, and the Gastrointestinal Symptom Assessment score in the intervention group was lower than that in the control group(P＜0. 01). The overall effective rate was 93. 33%(28/30) in the intervention group, which was significant higher than 73. 33%(22/30) in the control group(P＜0. 01). No serious adverse reaction was reported in either group. Conclusion The therapeutic apparatus for music therapy based on TCM syndrome differentiation is safe and effective in the treatment of functional abdominal distension. (目的 观察运用中医辨证移情音乐胃肠治疗仪改善功能性腹胀的效果。方法 将符合纳入标准的60例功能性腹胀患者随机分为两组, 各30例。对照组接受常规治疗和护理, 干预组在常规治疗的基础上采用中医辨证移情音乐胃肠治疗仪。两组均以7 d为1个疗程, 共2个疗程。记录与评价患者腹胀症状, 对比两组干预效果。结果 干预后, 两组胃肠症状评分均较干预前下降, 且干预组胃肠症状评分低于对照组, 差异有统计学意义(P＜0. 01)。干预组总有效率93. 33%(28/30), 高于对照组的73. 33%(22/30), 差异有统计学意义(P＜0. 05)。两组均无严重不良反应。结论 中医辨证移情音乐胃肠治疗仪改善功能性腹胀具有较好的疗效, 且安全可行。)

Due to the national requirements for keeping the surveying and mapping geographic information confidential, the sharing of long-distance pipeline coordinate data is always a difficulty for the application of informatization of the pipeline operators. Meanwhile, due to the release of the Provisions on the scope of state secrets in petroleum and petrochemical industry in 2013, great controversies were generated on the confidentiality classification of pipeline coordinate data. Hence, through the analysis on the recent cases for information security and the in-depth interpretation of the relevant national and industrial policies, the problem on confidentiality classification of pipeline coordinate data was clarified. In view of the pain points and business requirements in the current application of long-distance pipeline coordinate data, a technical scheme based on GCJ02 coordinate system was developed with reference to the relevant policies and technologies of the Ministry of Natural Resources of the People's Republic of China, and the management countermeasures were proposed in terms of the sense of security and confidentiality, map data and service, and business application, so as to meet the national regulatory requirements and the urgent needs of pipeline operators to deepen the application.(2 Figures, 1 Table, 27 References)

Trial difficulty prediction (TDP) is the task of automatically predicting the difficulty of a trial given the case text, which has a broad application prospect in judicial intelligent system. In practice, the tools of TDP rely heavily on the experience of experts, which leads different conclusions in predicting the difficulty of the trial. However, there are few related research work. To address these issues, this paper regards it as a text classification problem in natural language processing. Through the analysis, it is found that, traditional text classification methods don??t consider the structural uniqueness and logical dependence among trial elements in complaint, which makes it difficult to predict the difficulty of a trial accurately. In order to solve the mentioned challenges, this paper carefully studies indictments and considers the complex and simple trial elements for judging cases, presents an end-to-end model, MAT-TAN (mask-attention and topological association network). Specifically, this paper proposes a novel mask-attention network (MAT), to carry out fine-grained analysis of a case description text in indictments. The masking mechanism plays a role of the intelligent gatekeeper, focusing on the specific position of the trial elements in indictments. Together with the self-attention mechanism, it extracts the comprehensive and accurate characteristics of each trial element. This paper proposes a novel topological association network (TAN), which models the judicial logic dependency relationship between different elements, and effectively integrates the characteristics of different elements. Finally, the TDP is realized. The experimental results conducted on real-world datasets demonstrate that the MAT-TAN can improve the macro averaged F1 up to 0.036 compared with baselines, showing that it has a better performance in TDP.

Aiming at the solution to the time-consuming and low-precision disadvantage of present methodologies,the light field depth estimation method combining context information of the scene is proposed.This method is based on an end-to-end convolutional neural network,with the advantage of obtaining depth map from a single light field image.On merit of the reduced computational cost from this method,the time consumption is consequently decreased.For improvement in calculation accuracy,multi orientation epipolar plane image volumes of the light field images are input to network,from which feature can be extracted by the multi-stream encoding module,and then aggregated by the encoding-decoding architecture with skip connection,resulting in fuse the context information of the neighborhood of the target pixel in the process of per-pixel disparity estimation.Furthermore,the model uses convolutional blocks of different depths to extract the structural features of the scene from the central viewpoint image,by introducing these structural features into the corresponding skip connection,additional references for edge features are obtained and the calculation accuracy is further improved.Experiments in the HCI 4D Light Field Benchmark show that the BadPix index and MSE index of the proposed method are respectively 31.2% and 54.6% lower than those of the comparison me-thod,and the average calculation time of depth estimation is 1.2 seconds,which is much faster than comparison method.

The formation mechanism of tooth surface twist in helical gear generating machining （worm grinding and hobbing） and the measure to reduce the tooth surface twist are mainly studied. Based on the diagonal worm grinding or hobbing machining method， the tooth surface equation of helical gears with axial modification is constructed. The standard helical gear tooth surface and the helical gear machining tooth surface with axial modification are compared to obtain the correlation law between the speed ratio of axial feed and diagonal feed of the generating tool （worm or hob） and tooth surface twist. Based on the derived tooth surface equation of the helical gear with axial modification， Matlab and CATIA combined modeling technology is used to establish a three-dimensional geometric model of the helical gear pair with axial modification. The simulation analysis of the meshing performance of the gear pairs with different levels of tooth surface twist shows the influence of tooth surface twist on the meshing performance. The research shows that， the larger the helix angle of helical gear with axial modification is， the more serious the tooth surface twist is. Modification amount does not affect the twist of the tooth surface. The ratio of the axial feed and diagonal feed of the tool has the best match with the tooth surface twist. Improved tooth surface twist can effectively reduce the transmission error and improve the gear carrying capacity. The CAD/CAE integrated modeling and analysis method based on the combination of Matlab， CATIA and finite element commercial software provides methods and tools for the research on tooth surface twist of helical gears with axial modification.

Objective To investigate the non⁃invasive evaluation of intracranial pressure (ICP) by transcranial Doppler ultrasound (TCD) in patients with traumatic brain injury (TBI). Methods The clinical data of 68 patients with TBI were retrospectively analyzed. Pulsatility index (PI) and optic nerve sheath diameter (ONSD) were performed by TCD, simultaneous recording of ICP parameter one week after surgery. The relationship between ONSD, PI and ICP was analyzed by Pearson correlation analysis. The area under the curve (AUC) was used to evaluate the diagnostic sensitivity of ONSD and PI for prediction intracranial hypertension. Results One week after surgery,ICP was positively correlated with ONSD (r= 0.679, P=0.000). There was also a positive correlation between ONSD and ICP ≥ 20 mm Hg (r=0.665, P= 0.000) and ICP

There is no analytical calculation method and formula for the calculation of the bending strength of the face gear.For this reason，based on the principle of tooth surface contact analysis，the formula for calculating the length of the bending arm is derived，reference the calculation standard （ISO 10300（3）：2014） of bevel gear bending strength，an analytical calculation method for the root bending strength of non-orthogonal helical face gear is proposed.Taking a parameter face gear transmission pair as an example，the Abaqus finite element software is used to calculate the root bending strength，and the root bending stress value of the non-orthogonal helical face gear is extracted.The finite element results are compared with the analytical calculation results，the error is about 8.5%.The results show that the calculation method of the calculation method of the root bending stress of non-orthogonal helical face gears is correct and feasible.The problem that the calculation of the bending strength of the face gear has no analytical formula is solved.After refinement and perfection，it is expected to form a basic calculation formula for the bending strength design of the face gear.

In order to further understand the pyrolysis mechanism of decabromodiphenyl ethane (DBDPE) and the evolution process of its main products, the thermal degradation reaction mechanism of DBDPE was studied using density functional theory (DFT) method M06-2X/6-311G(d,p). Possible reaction paths for DBDPE pyrolysis were designed, and the kinetic and thermodynamic parameters of various reaction pathways were calculated. The results show that the initial pyrolysis of DBDPE is dominated by the fracture of aliphatic H2C--CH2 bond, which results in the formation of a large number of pentabromobenzyl radicals. The bromine radical produced by Caromatic--Br bond fracture is a competitive reaction channel, and the bromine radical can further react with the pentabromobenzyl radical to form pentabromobenzyl bromide. The pentabromobenzyl radicals and bromine radicals produced in the initial reaction can promote the decomposition of DBDPE, which are mainly reflected in the abstraction of H atoms from aliphatic H2C--CH2 bonds with energy barriers of 21.9 kJ/mol and 38.3 kJ/mol, respectively, resulting in the formation of pentabromotoluene, pentabromostyrene, hexabromobenzene, and hydrogen bromide. In the pyrolysis reaction processes of DBDPE with hydrogen radical, the addition of hydrogen radical reduces the reaction energy barrier of DBDPE pyrolysis, and the reaction energy barrier of adding hydrogen radical to the aromatic-carbon of Caromatic--CH2 of DBDPE is the lowest (17.9 kJ/mol). The main products of co-pyrolysis of DBDPE with H radicals are hydrogen bromide, polybrominated diphenylethane, and brominated monoaromatic compounds. [ABSTRACT FROM AUTHOR]

[Objectives]In the fields of marine oil exploitation and lake water quality monitoring,in order to realize the self-positioning and self-navigation of ships,it is necessary to realize the velocity control for dynamic positioning ship.[Methods] Ship motion model of 3 Degree of Freedom (3-DOF) is established,and constant-velocity navigation control algorithm based on fuzzy decoupling control is proposed. Fuzzy controllers are designed separately for surge,sway and yaw loop to solve the modeling error of controlled object. In order to solve the coupling problem between sway loop and yaw loop variables,feed-forward compensated decoupling scheme is designed to eliminate mutual effect between controlled variables.[Results] The experimental results of the constant-velocity navigation control simulation platform show that the fuzzy decoupling control algorithm has good dynamic performance,high steady-state accuracy and certain robustness.[Conclusions]The study results provide references for the precise velocity control for dynamic positioning ship.

Objective To analyze the clinical features of multiple spontaneous intracerebral hemorrhages (MICH). Methods Conservative therapy, puncture and drainage, hematoma removal and/or decompressive craniectomy were used in the treatment of 630 intracerebral hemorrhage (ICH) patients, who were divided into 2 groups: 30 cases with MICH and another 600 cases with solitary intracerebral hemorrhage (SICH). Three months after onset, modified Rankin Scale (mRS) was used to evaluate the prognosis of all cases. Results Compared with patients in SICH group, the occurrence rate of hypertension > 5 years (P = 0.008), diabetes mellitus (P = 0.024), hypercholesterolemia (P = 0.050) and previous ischemic stroke (P = 0.026) were all significantly higher in MICH group. The mean arterial pressure (MAP) level (P = 0.002) and the incidence of limb movement disorder (P = 0.000) were significantly higher in patients with MICH than those with SICH. Basal ganglia and thalamus were the predilection sites of hematoma (P = 0.001). Patients with MICH had worse prognosis compared to those with SICH 3 months after onset (P = 0.006). Conclusions Hypertension > 5 years, diabetes mellitus, hypercholesterolemia and ischemic stroke were identified to be the pathophysiological basis of MICH in this study. All patients with MICH had more serious clinical manifestations after onset and worse prognosis. DOI: 10.3969/j.issn.1672-6731.2016.01.008

DOI: 10.3969/j.issn.1672-6731.2017.03.013

The synthesis of N, N'-l,4-bis( 4-methylphenyl) sulfonyl-liomopiperzine was studied with N-methylpyirolidone as solvent, NaOH as acid binding agent and 1,3-dichloropropane as cyclization reagent. The effects of reaction time, the molar ratio of reactants, reaction temperature, and the amount of sodium hydroxide on the yield of N,N'-1,4-bis(4-methylphenyl) sulfonyl-homopiperzine were investigated. The result showed that the order of effects was as follows: alkali amount > temperature > molar ratio of reactants. The yield could reach 81.93% under the conditions of 5 mmol N,N-bis(toluenesulfonyl)-1,2-ethylenediamine, 7.969 mmol 1,3-dichloropropane and 0.548 g sodium hydroxide,123 °C react 40 min. The kinetic analysis of the synthesis of N,N-1,4-bis(4-methylphenyl) sulfonyl-homopiperzine was carried out at 115, 125, 135 °C according to the second-order reaction mechanism, and the activation energy of this reaction was 76. 233 kJ/mol. [ABSTRACT FROM AUTHOR]

Slow water bodies, mainly represented by landscape lakes and ecological wetlands, are becoming more and more important in urban construction. How to increase water exchange areas and slow down the deterioration of water quality is an important problem we are faced with. Based on particle image velocimetry (PIV) technique, the effects of spacing ratio, diversion flow rate and shape of double bluff body (islands) on two-dimensional flow field distribution, water displacement rate and average velocity of the flow field in slow water are studied. The experimental results show that the diversion flow rate is the main controlling factor for the intensity of the flow field in the slow water, and the hydrodynamic exchange efficiency and the average velocity of the water flow are proportional to the diversion flow. The bluff body can obviously improve the range of flow movement and increase the fluidity of water. It is found that the fluidity of the flume water is the strongest and the displacement rate is the highest when the prism and cylinder are disturbed together. The maximum displacement rate reaches 72.89% when Q=6.8 L/min and G/D=6. The experimental results will provide some reference for optimizing the layout of islands in the landscape lake. [ABSTRACT FROM AUTHOR]

The distribution of group-specific component(G) types was examined by disc electrophoresis from 508 unrelated residents in Wuhan. Discontinuous buffer system was used in this investigation. The polyacrylamide concentration used for separating gel was 9%, used for concentrating gel was 3.125％. After electrophoresis the gel was stained with amidoblack and destained by electrilytic destaining. This method shortened destaining time compared with the conventional destaining method. The distribution of Gc types in the population of Wuhan was as follows: Gc 1—1=255 (50.2%), Gc2-1=214 (4 2.1%), Gc2-2=39 (7.7％). The gene frequencies were calculated at Gc=0.7126. Gc 2=0.2874.

Objectives: The purpose of this study was to determine students' knowledge, attitudes, and behavior regarding the correct use of analgesics and to examine the related factors. Methods: A probability-proportionate-to-size sampling method was used. Thirty-three senior/vocational high schools were randomly selected. A total of 2,910 students completed the online selfadministered questionnaire in 2013. Results: Greater than one-half of the students did not know that when taking pain medication containing acetaminophen, adults should not exceed 4,000 mg per day. Approximately 22.1% of the students were unaware of the hepatotoxicity risk associated with acetaminophen. Multiple regression analysis showed that students in higher grades who had greater levels of correct analgesic knowledge, attitude, and self-efficacy, take analgesic prescriptions and use other pain management techniques, such as hot/cold therapy, relaxation, and massage, and were more likely to demonstrate correct analgesic use behaviors if they had received pain management information from family members and had read the warning labels. In contrast, students taking analgesics from drug stores or family/friends were more likely to practice poor analgesic use behavior. Conclusions: The promotion of partnerships between schools and pharmacists is suggested, along with the implementation of teachers' training workshops, students' education programs, and parent-child activities in order to enhance correct analgesic use literacy. [ABSTRACT FROM AUTHOR]

Castanopsis fissa is an important pioneer species commonly used in the restoration of degraded subtropical grassland in southern China. The study on the effects of litter coverage and watering frequency on the seed germination and seedling survival of C. fissa showed that the effects of litter coverage depended on soil moisture condition. When the watering frequency was 1 time per day, litter covering could restrain seed germination and increase seedling mortality; while watering 2 or 3 times per day, the covering would ameliorate soil moisture condition, and benefit seed germination and seedling establishment. The biomass of C. fissa seedlings was enhanced significantly by litter covering. it was suggested that keeping a litter layer on soil surface could benefit the seed germination and seedling survival of C. fissa to restore subtropical shrub land. [ABSTRACT FROM AUTHOR]

Objective: To construct the stable transfection Hela cell lines with lentiviral vector which may interfer the expression of ALDH-1,and to further explore the biological characteristics between the expression of ALDH-1 and Hela cell lines. Methods: Hela cells were cultured in vitro, added the different concentration of puromycin to screen, and to determine the best screening concentration; Hela cells were tested from 24-well plates, threw away the old culture medium, and added 400 µL of new medium to the plates, and then added four kinds of lentivirus solution) ALDH-1, ALDH1-1719, ALDH1-740, ALDH1-921,and there was a kind of lentivirus which has the best interfering effect) and one kind of negative control lentivirus solution (LV3-NC) to the plates, at last added Polybrene liquid to them. Every lentivirus solution volume was added about 50 µL (each of viral titers were 1x10-8 TU/mL), and the Polybrene was added about 0.5 µL (concentration of 5 mg/L) respectively. Fresh culture medium was changed after 24 hours, and observe the fluorescence after 72 hours. At last, using the best screening concentration ofpuromycin to screen the transfection Hela cells, and to get the stable transfection cell lines. Results: The best screening concentration of puromycin was 0.9 mg/L, lentiviral vectors were successfully transfected into Hela cells, and the stable transfected cell lines were successfully obtained after two weeks. Conclusions: Hela cells which has stable interference of ALDH -1 were successfully constructed. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of electroacupuncture(EA) stimulation on proliferation and diffe-rentiation of endogenous neural stem cells as well as Jagged1/Notch1 pathway in AD model mice, so as to explore its mechanism underlying amelioration of AD., Methods: A total of 40 6-week-old male APP/PS1 transgenic AD mice were randomly divided into EA group ( n =20) and AD model group ( n =20), and other 20 normal C57BL/6J mice of the same age were used as the normal control group. The mice in the EA group received EA (10 Hz, 2 mA) at "Baihui"(GV20), "Fengfu"(GV16) and bilateral "Shenshu" (BL23) for 20 min, once daily, 6 days a week for 16 weeks. The mice's learning-memory ability was detected by Morris water maze tests. The Aβ senile plaques in the hippocampal CA1 region were detected by Congo red staining, the immunofluorescence double label of BrdU, neuronal nuclear antigen (NeuN) and astrocyte specific protein GFAP in dentate gyrus of hippocampus was performed for detecting the proliferation and differentiation of the endogenous neural stem cells. The expression levels of Nestin (neuron specific protein) and GFAP were detected by Western blot, and those of Jagged1 and Notch1 mRNAs and proteins in the hippocampus were detected by real-time fluorescence quantifative PCR and Western blot., Results: Compared with the normal control group, the escape latencies at 2 nd , 3 rd and 4 th day, and Aβ senile plaques were significantly increased ( P <0.05, P <0.01), whereas the platform crossing times and time spent in the target quadrant, the expression levels of Jagged1 mRNA and Nestin protein were remarkably down-regulated ( P <0.05) in the model group. Following EA intervention, the escape latencies at the 3 rd and 4 th day, Aβ senile plaques, immunofluorescence density of BrdU/GFAP, and GFAP protein expression were pronouncedly decreased ( P <0.05, P <0.01), while the platform crossing times, platform quadrant residence time, immunofluorescence density of BrdU/NeuN, expression levels of Jagged1 and Notch1 mRNAs and proteins and Nestin protein evidently increased ( P <0.05, P <0.01), suggesting an enhancement of proliferation and diffe-rentiation of endogenous neural stem cells into neurons and a suppression of the proliferation and differentiation towards astrocytes in the hippocampus., Conclusion: EA at GV20, GV16 and BL23 can improve the learning-memory ability, promote the proliferation and differentiation of endogenous neural stem cells towards neurons and inhibit the proliferation and differentiation of endogenous neural stem cells towards astrocytes in the hippocampus, which may be achieved by regulating Jagged1/Notch1 pathway.

Objective: To observe the effect of electroacupuncture (EA) on the proliferation of endogenous neural stem cells in the hippocampus of young mice with Alzheimer's disease (AD), so as to explore its mechanisms underlying improvement of AD., Methods: Forty 1.5-month-old APP/PS1 transgenic male mice were randomly divided into an EA group and a model group, 20 mice in each group, and other 20 C57BL/6J male mice of the same age were used as the normal control group. EA (intermittment wave 10 Hz, 2 mA) was applied to "Baihui" (GV 20), "Fengfu" (GV 16) and "Shenshu" (BL 23) for 20 min, once a day, 6 days a week for 16 weeks. H.E. staining was used to assess histopathological changes of neurons of the hippocampal dentate gyrus. Immunohistochemical stain was used to detect the expression of 5-bromodeoxyuridine (BrdU)-positive in the hippocampus, and immunofluorescence double-labeled technique was used to detect the number of proliferated positive neurons of hippocampal neural stem cells. The expression levels of brain derived neurotrophic factor (BDNF) and Nestin mRNA and protein were detected by using real-time PCR and Western blot, separately., Results: The immunoactivity of BrdU, and the expression levels of BDNF and Nestin mRNA and protein in the hippocampus in the model group were significantly lower than in the normal control group ( P <0.01, P <0.05), and considerably higher in the EA group than in the model group ( P <0.01, P <0.05). The number of BrdU/NeuN dual labeled neurons was slightly increased in the model group than in the normal control group ( P >0.05), and evidently increased in the EA group relevant to the model group ( P <0.05), suggesting a proliferation of hippocampal neural stem cells. After modeling, the neurons of hippocampal dentate gyrus were arranged loosely and irregularly and their structure was fuzzy, with an appearance of different degrees of nuclear pyknosis, whereas in the EA group, the neuronal contour was clear and the nuclear structure was relatively distinct., Conclusion: EA can activate the proliferation of neural stem cells in the hippocampus in AD mice, which may contribute to its function in improving the neuronal structure by upregulating the expression of BDNF.

Objective: To explore the cause for the failure of non-invasive prenatal testing (NIPT) and feasibility of repeated testing., Methods: Clinical data, test results and pregnancy outcomes of 40 311 pregnant women who received NIPT test from January 2011 to December 2018 were reviewed., Results: Among all the pregnant women, 1116 cases failed in the first test, 9 cases (0.81%) had fetal free DNA concentration lower than 4%, 663 cases (59.41%) were retested after the establishment of Z value gray area, and the remainder 444 cases (39.78%) needed to be retested after the blood collection due to the fetal free DNA concentration lower than 4%. After retesting, 1069 cases (95.78%) obtained effective NIPT results. The results showed that 53 cases were at high risk (6 cases for trisomy 21, 6 cases for trisomy 18, 13 cases for trisomy 13, 16 cases for sex chromosomal abnormality, 12 cases for chromosomal copy number variation). Forty-eight cases were selected for invasive prenatal diagnosis, and 2 cases of 47, XXY and 2 CNV were confirmed. A total of 47 cases (0.12%) did not obtain results because the concentration of fetal free DNA was lower than 4%. Only 16 cases (34%) chose invasive prenatal diagnosis., Conclusion: Repeated detection of the gray area of Z value can reduce the false positive rate of NIPT and invasive prenatal diagnosis, and the feasibility of repeated detection is high. In the case of fetal free DNA concentration lower than 4%, the success rate of obtaining effective NIPT results by re-sampling and re-detection increases with the increase of gestational age, but may delay the diagnosis for fetal aneuploidies. Therefore, personalized estimation should be made according to gestational age and clinical indications. It is suggested that pregnant women should choose invasive prenatal diagnosis when they have failed in the retest.

The ecological security pattern aims to maintain the integrity of landscape pattern and the continuity of ecological process to ensure urban ecological security. Current ecological source identification methods lack the consideration for ecological land degradation risks and human ecological needs. Taking Wuhan City as an example, the comprehensive value of ecological land was calcula-ted from three aspects, including ecological land quality, ecological degradation risk, and ecological demand to identify ecological source areas. Basic ecological resistance surfaces were constructed based on land use types and nighttime light data. Potential ecological corridors were identified based on the minimum cumulative resistance model. Ecological "pinch points" were recognized based on circuit theory. After that, we built a "four horizontal, three vertical and ten groups" ecological safety pattern in Wuhan. The results showed that the area of ecological land in Wuhan was 2138.2 km 2 , accounting for 24.9% of the total area of the city. These areas were primarily attributed to waters and foress, clustering in the north and south of the city. The total length of ecological corridors was 1222.42 km, with 566.75 km aquatic corridors and 655.67 km land corridors. The aquatic corridors traversed the city area to form a double cross-shaped pattern. The land corridors circularly distributed around the city. Comprehensively, the "four horizontal and three vertical" spatial pattern had appeared. There were a total of 44 ecological pinch points on the ecological corridor, distributed circularly with the central city as the kernel. The extant protection space had almost been included in the identified ecological source area, supporting the ecological significance of the identification framework. It would help to provide a quantitative framework for the construction of ecological secu-rity pattern in the metropolitan area and to guide the relevant urban spatial planning.

Objective: To characterize the prevalence and species/genotypes of Cryptosporidium spp. in farmed pigs in the north of the Yangtze River in Anhui Province., Methods: A total of 500 samples of pig feces were obtained from seven largescale pig farms in the north of the Yangtze River in Anhui Province. PCR and sequences analysis of the small subunit rDNA gene were used to detect and identify the Cryptosporidium species/genotypes., Results: The overall prevalence of Cryptosporidium was 4.8% (24/500). Additionally, Cryptosporidium prevalence was 40.0% in Qianshan and 6.3% in Chuzhou, respectively. No Cryptosporidium infection was found in other sampling areas. The DNA sequence analysis of the SSUrDNA gene revealed that all of the isolates represented C. scrofarum . The Cryptosporidium infection rate (9.1%) of pigs (> 60 days) was significantly higher than the rates of both pigs (< 30 days) and pigs (30-60 days) (both P < 0.01)., Conclusions: C. scrofarum in the farmed pigs in the north of the Yangtze River in Anhui Province may be a source of Cryptosporidium infection and pose a potential public health threat to humans and other animals, and therefore, the status should be paid more attention to.

Objective: To investigate the influence of phenylalanine(Phe) on glucose uptake in mouse myoblast cell line C2C12 and to explore its relationship with mTOR-p70S6K pathway., Methods: C2C12 cells were cultured to promote formation of multinucleated myotubes in vitro. The cells were deprived and incubated with Phe at different concentrations (1.25,2.5,5,10,20 mmol/L).Krebs-Ringer buffer (KRB) was used as control.The 2-NBDG was used to measure glucose uptake of C2C12.The expression of mTOR,p70S6K,IRS-1,and Akt protein were evaluated by Western blot., Results: Compared with KBP treatment,glucose uptake of the cells incubated with 5 mmol/L leucine (Leu) was decreased by 30% ( P =0.001), while a 40% increase was detected in the cells incubated with 5 mmol/L Phe ( P <0.01).The promotion of glucose uptake was Phe concentration-dependent.Phe stimulation had no effect on the phosphorylation of mTOR at Ser2448. Phosphorylation of p70S6K at Thr389 was inhibited in the cells incubated with Phe at concentration higher than 1.25 mmol/L, but the difference was not significant ( P =0.815). Leu stimulated but Phe over 1.25 mmol/L inhibited phosphorylation of IRS-1 at Ser636/639, although the difference was not significant ( P =0.381).Neither Leu nor Phe affected the expression of phospho-Akt (Ser473) significantly., Conclusion: Phenylalanine inhibits phosphorylation of IRS-1 at Ser636/639 possibly through inhibiting the activation of p70S6K.The effect of Phe on mTOR-p70S6K pathway is Akt-independent., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Objectives: To study the association of nt3434 A&#8594;G mutation in mitochondrial DNA NADH dehydrogenase 1 subunit ( ND 1) gene with diabetes mellitus., Methods: PCR-RFLP was used to detect the nt3434 A&#8594;G variant of mtDNA ND 1 gene in 216 diabetic patients and 203 healthy control individuals. Characteristics of mutation and clinical indicators in nt3434A&#8594;G family were analyzed., Results: nt3434A&#8594;G mutation was detected in one diabetic patient but not found in NC group. This patient had low insulin secretion, low BMI, and elevated serum lactate acids. No significant difference was found in the mutation frequencies between these two groups. nt3434 A&#8594;G mutation was also detected in this patient's sister and daughter, who were normal glucose tolerance and had slightly elevated serum lactate acids levels., Conclusions: Further investigation would be helpful to answer whether nt3434A&#8594;G mutation of mitochondrial DNA ND 1 gene is associated with an increased risk of diabetes.

Objective: To detect chromosomal imbalance in a fetus with complex congenital heart disease, and to correlate the genotype with the phenotype., Methods: Routine G-banding was carried out to analyze the karyotypes of the fetus and its parents, and single nucleotide polymorphisms array (SNP-array) was used for delineating fine genomic aberrations. The detected aberrations were confirmed with multiplex ligation-dependent probe amplification (MLPA)., Results: The fetus and its parents all showed a normal karyotype, while array-SNP has detected a 13.87 Mb duplication at 4p16.3-p15.33 and a 15.65 Mb deletion at 11q23.3-q25 in the fetus. The results were confirmed by the MLPA assay., Conclusion: The partial trisomy 4p (Wolf-Hirschhorn syndrome) and partial monosomy 11q (Jacobsen syndrome) probably underlie the complex heart defects detected in the fetus. Analysis of the karyotypes of its parents offered no help for the determination of the aberrant type and recurrent risk. Compared with routine karyotype analysis, aberrant regions can be identified with array-SNP with greater resolution and accuracy. This has provided useful information for prenatal diagnosis and genetic counseling.

Objective: To search for new potential diabetogenic mtDNA defects by scanning mtDNA genome in mitochondrial DNA diabetes (MDM) pedigrees., Methods: Blood samples were collected from the family members in two suspected MDM pedigrees, which were both maternal transmitted and early-onset diabetes. The whole mtDNA genome except D-loop was detected by direct sequencing in probands of these two diabetic pedigrees. Novel mutations displayed by direct sequencing were then screened in 200 normal glucose tolerant controls and 100 early-onset diabetic patients., Results: We found a novel nt14319T --> C mutation in No. 2 pedigree, but no pathogenic mutation was found in No. 1 pedigree. The mutation of nt14319T --> C, which not being reported previously, locate in the region of ND6 subunit, causing amino acid change (asparagine --> aspartic acid). The frequencies of 14319T/C substitution in 100 early-onset diabetic patients and 200 control subjects were 6% and 5%(P > 0.05)., Conclusion: A novel mutation 14319T --> C was identified in No. 2 pedigree. All three diabetic family members in this pedigree haboured 14319T --> C mutation, indicating that it may be the major pathogenic mutation for this family.

Objective: To examine the association of the Thr394Thr polymorphism of PPARGC1A gene with type 2 diabetes (T2DM), insulin resistance (IR) and other metabolic disorders in a Chinese population., Methods: Three hundred and seven subjects including 151 T2DM patients and 156 normal glucose tolerant controls (NC) were enrolled in this study. The Thr394Thr G/A polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Glucose, insulin, lipids levels were determined in all subjects. Body mass index (BMI), waist circumferences, index of homeostasis model assessment-insulin resistance (HOMA-IR) and blood pressure were also measured., Results: The diabetic subjects had higher levels of BMI, waist circumferences, blood systolic pressure, triglycerides and lower levels of high density lipoprotein-cholesterol (HDL-C) compared with those of control subjects (P<0.05). About 43.7% (66/151) of the T2DM subjects had the AG genotype, while 37.2% (58/156) in the NC group. The frequency of the A allele was 0.225 in T2DM, and 0.186 in the NC subjects. There were no significant differences either in genotype or allelic distribution of G/A polymorphism between the two groups. In the T2DM group, subjects with AA and GA genotypes had significantly higher levels of HOMA-IR, waist circumferences and lower levels of HDL-C (P<0.05) than those carrying GG genotype. HOMA-IR in subjects with AA and AG were significantly higher than those with GG genotype in both groups., Conclusion: The A allele of the Thr394Thr (G-->A) polymorphism of the PPARGC1A gene was associated with insulin resistance, and may be related to central obesity and decreased HDL-C levels in Chinese population. The relationship between this polymorphism and T2DM needs further investigation.

This investigation was directed to the metabolic syndrome and the islet beta-cell secretory function in the first-degree relatives (FDR) of type 2 diabetic patients in Sichuan province. A large cohort study was designed. Totally 1929 subjects were investigated. They were in two groups: FDR group comprising 505 first-degree relatives of type 2 diabetic patients, and Control group comprising 1424 controls without positive family history of Diabetes. Blood pressure, weight, waist, plasma glucose, lipids and insulin were measured. HOMA-IR and HOMA-beta indexes were used to evaluate insulin resistance and beta-cell secretion function. The insulin sensitivity index (ISI) and glucose disposition index (DI) were also used to evaluate insulin resistance. After adjustment for age and sex, HOMA-IR increased, ISI, DI and HOMA-beta decreased in FDR group when compared with controls (P < 0.05). The incidence of co-existed three or more metabolic disorders and metabolic syndrome was higher in FDR group than that in control group (P < 0.05). In FDR group, HOMA-IR increased, HOMA-beta, DI and ISI decreased while the number of co-existing metabolic disorders increased. But when the number of co-existing metabolic disorders > or = 4, HOMA-IR increased no longer and ISI decreased no more. Metabolic disorders occurred more frequently in FDR of diabetic patients than those in individuals without positive family history. As the number of co-existing metabolic disorders increased, the beta-cell secretion function and insulin sensitivity became worse. Our study indicated that it is necessary to keep on monitoring the metabolic index in FDR of type 2 diabetes and provide early preventive interventions.

Objective: To explore the surgical extent and to improve the surgical techniques of the Piver class III hysterectomy on invasive cervical cancer, so as to reduce the urinary tract complications, shorten the surgical duration, decrease the hemorrhage and blood transfusion., Methods: The study group, 196 cases with stages Ib and IIa carcinoma of the cervix underwent the modified Piver class III hysterectomy from June 2000 to May 2005. The control group, 176 cases of the same stages underwent the Piver class III hysterectomy between June 1994 and May 1999. The modified Piver class III hysterectomy mainly include the surgical extent and some surgical techniques as follows. The cervicovesical and vesicovaginal space are separated with assistance of electrotome. Half of the uterosacral ligaments are removed with electrotome. The tunnel of the ureters is separated and penetrated or not. The anterior leaf of the cervicovesical ligaments is removed and the uterine artery are removed at the same time, while the ureter branch from the uterine artery are preserved. When the ureters are drawn to the lateral side of the body with an "S" hook and the urocyst lateral recessus are expanded, the cardinal ligaments can be exposed and be removed of 3/4. But part of the inferior of these ligaments should be preserved. The paracolpium are resected about 2 cm, 2 - 3 cm tissue of the vagina is removed., Results: Compare with the control group, the urinary tract complications of the study group were significantly reduced (51.1% versus 23.0%, P < 0.01). There were a shorter surgical duration [(132 ± 20) min], less of the hemorrhage [(322 ± 100) ml] and blood transfusion [(154 ± 79) ml] in the study group than those in the control group (all P < 0.05). While, there was no significant difference at the survival rates of 5 years between the two groups (87.8% versus 88.6%, P = 0.793)., Conclusion: The modified Piver class III hysterectomy is effective and applicable for patients with cervical cancer.

Objective: To screen the mutation of the beta and gamma subunits of epithelial sodium channel gene SCNN1 in two families with Liddle's syndrome., Methods: Two patients clinically diagnosed as Liddle's syndrome and their family members were enrolled. Peripheral blood samples were collected and total genomic DNA was prepared. Polymerase chain reaction (PCR) was used to amplify the exon 13 of the SCNN1B and SCNN1G gene. PCR products were purified and subjected to direct DNA sequencing., Results: A heterozygous nonsense mutation at codon 564 of the SCNN1B gene from CGA(Arg) to stop codon(TGA) was detector in the proband of family 1. More importantly, a novel heterozygous nonsense mutation of CAG(Gln) to stop codon TAG at codon 567 of the SCNN1G gene was detected in the proband and another two members of family 2., Conclusion: Screening for specific mutations of the SCNN1 gene in relatives of patients with Liddle's syndrome can be used to identify the previously unrecognized cases within the family. A new nonsense mutation(Q567X) of the SCNN1G gene is likely the cause of Liddle's syndrome in family 2.

Objective: To investigate the characteristics of glycemic excursions in people with normal glucose tolerance (NGT) in Chengdu., Methods: A total of 50 non-obese people with normal glucose tolerance (NGT, 23-68 years old), normal blood pressures and lipid profiles participated in the study. The fluctuations of glucose levels in the participants were measured by a continuous glucose monitoring system (CGMS) for three days 72 h. The 48 h mean blood glucose (MBG), mean amplitude of glycemic excursions (MAGE), Largest amplitude of glycemic excursions (LAGE), Postprandial peak glucose (PPG), Postprandial glucose excursion (PPGE), Mean of postprandial glucose excursion (MPPGE), and absolute means of daily differences (MODD) were measured., Results: The number of glucose values detected by CGMS amounted to 861+/-7 with a mean absolute difference (MAD) of 11.3%+/-10.6%. The CGMS values were significantly correlated with the capillary glucose measurements (n=1076, r=0.761, P<0.005). The participants had a MBG of (5.9+/-1.2) mmol/L, a MAGE of (1.7+/-0.7) mmol/L, a LAGE of (4.4+/-1.9) mmol/L, a daily glycemic peak (PPG) of (8.7+/-1.7) mmol/L, a nadir level of (4.3+/-0.7) mmol/L, a MPPGE of (2.3+/-1.6) mmol/L, and a MODD of (0.75+/-0.79) mmol/L. The post-breakfast Postprandial glycemic excursions (PPGE) were lower than those of post-lunch and post-dinner (P=0.01 and P=0.05). The postprandial glucose excursions in the 60-70 year-old participants were the highest (P<0.022). In 95% (77%-100%) of the daytime, the glycose levels fluctuated between 4.1 and 8.8 mmol/L, and 78% of the participants (n=39) had hyperglycemia (BG>7.8 mmol/L) and 10% (n=5) had asymtomatic hypoglycemia (BG<2.8 mmol/L)., Conclusion: CGMS tests may be important for detecting asymptomatic hyperglycemia and hypoglycemia. The NGT people in Chengdu have exhibited abnormal blood glucose values in CGMS, revealing problems in people with normal range of blood glucose.

Objective: To investigate the mechanism of autologous platelet-rich gel (APG)in the treatment of refractory diabetic dermal ulcers., Methods: After the treatment of refractory diabetic dermal ulcers with APG at 0, 3, 6, 9, 15 days, the protein levels of PDGF-BB, VEGF, IGF-1, EGF and TGF-beta1 in the granulation tissue were detected by ELISA, while the dimensions of ulcer area were measured at the same time., Results: The areas of ulcers were obviously reduced at the third and fifteen day after APG treatment (P < 0.05). The concentrations of these 5 growth factors in the granulation tissue were began to increase after 3 days treatment, the peak of PDGF-BB emerged at the third day (P < 0.05), and the peaks of VEGF, IGF-1, TGF-beta1 were found at the ninth day (P < 0.05). The concentration of VEGF increased 2.1-fold, IGF-1 increased 1.95-fold, EGF increased 1.75-fold, PDGF-BB increased 1.89-fold and TGF-beta1 increased 1.67-fold., Conclusion: The expression of multiple growth factors are increased in granulation tissue of refractory diabetic dermal ulcers after the treatment of APG,which might be one of the mechanism of APG to treat refractory diabetic dermal ulcer.

Objective: To investigate the relationships between Paraoxonase1 (PON1) gene L55M polymorphism and coronary heart diseases (CHD) complicated with fasting hyperglycemia (FHG) or type 2 diabetes mellitus (T2DM)., Methods: Two hundreds and eighty four patients were recruited in this study, which comprised 86 patients with CHD, 62 with CHD and FHG, 46 with CHD and T2DM, and 90 patients without the above conditions. The sequences of the PON1 gene L55M polymorphism were analysed by the SNPs methods., Results: The frequency of allele L was 0.86. No significant differences in the frequencies of the three genotypes (LL, LM, and MM) were found among the four groups of patients. The L allele of PON1 L55M was not associated with the number and range of coronary arterial lesions (chi2 = 0.56, P = 0.76; chi2 = 0.11, P = 0.74, respectively)., Conclusion: PON1 gene L55M polymorphism exists in Han populations in Guangdong, with LL as the main genotype. PON1 L55M gene polymorphism and allelic genes are not associated with the level of fasting blood glucose or diabetes mellitus. There is no evidence to support the relationship between PON1 L55M gene polymorphism and coronary artery lesions.

Objective: To investigate the association between L296Q polymorphism in the cholesterol ester transfer protein(CETP)gene and type 2 diabetes mellitus(T2DM)and blood lipids., Methods: Plasma glucose and lipid levels were measured in a total of 303 subjects recruited from the West China Hospital of Sichuan University. The subjects were divided into 4 groups according to the levels of plasma glucose and triglyceride, namely T2DM with hypertriglyceridemia group, group of T2DM with normal triglyceride, group of hypertriglyceridemia without DM and group of normal controls, respectively. Genotypes of L296Q polymorphism in the CETP gene of all subjects were analyzed by real-time fluorescent quantitative PCR(FQ-PCR)., Results: No significant differences were observed in the frequencies of genotypes LL and LQ and the 296Q allele among the four groups (chi-square=3.459, P>0.05; chi-square=3.155, P>0.05, respectively), nor the frequencies of genotypes LL and LQ between the T2DM and non-T2DM, or plasma lipid levels between the 296Q allele carriers and those of genotype LL., Conclusion: No association was found between the L296Q polymorphism in the CETP gene and T2DM as well as plasma lipid levels in various groups of Chinese in this study.

Objective: To construct and express the recombinant human adiponectin (gAd) global domain., Methods: gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG., Results: The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis., Conclusion: The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

Objective: To compare the platelet enrichment ratio of platelet-rich plasma (PRP) prepared by different centrifuge methods and to compare the concentration of growth factors released from autologous platelet-rich gel (APG) with the whole blood., Methods: Thirteen diabetic patients with refractory skin lesions were enrolled in APG treatment. (1) Three kinds of centrifuge methods were selected for PRP by 11 diabetic patients: A (n = 6): 529 x g for 4 minutes in the first centrifuge and 854 x g for 6 minutes in the second centrifuge; B (n = 5): 313 x g for 4 minutes in the first centrifuge and 1,252 x g for 6 minutes in the second centrifuge; C (n = 5): 176 x g for 5 minutes in the first centrifuge and 1,252 x g for 5 minutes in the second centrifuge. Platelet counted on the whole blood and PRP was determined. The APG, produced by combining the PRP with thrombin and calcium gluconate (10:1) was used by patients. (2) PDGF-BB, TGF-beta1, VEGF, EGF, and IGF-1 were measured in the APG and the whole blood using the enzyme-linked immunosorbent assay method., Results: (1) The average platelet concentration was higher in group B [(1,363.80 +/- 919.74) x 10(9)/L] than in groups A [(779.67 +/- 352.39) x 10(9)/L)] and C [(765.00 +/- 278.78) x 10(9)/L] and the platelet recovery rate was 75.2% +/- 21.0% in group B. (2) The concentration of growth factors all increased with the increasing platelet number. On average, for the whole blood as compared with APG, the PDGF-BB concentration increased from (145.94 +/- 133.24) pg/mL to (503.81 +/- 197.86) pg/mL (P < 0.05); TGF-beta1 concentration increased from (3.31 +/- 2.27) ng/mL to (5.67 +/- 4.80) ng/mL (P < 0.05); IGF-1 concentration increased from (14.54 +/- 35.34) ng/mL to (110.56 +/- 84.36) ng/mL (P < 0.05); and EGF concentration increased from (160.73 +/- 71.10) pg/mL to (265.95 +/- 138.43) pg/mL (P < 0.05). No increase was found for VEGF (P > 0.05). (3) There was positive correlation between the platelet concentration and PDGF-BB and TGF-beta1 (r = 0.627, r = 0.437, P < 0.05). (4) Thirteen diabetic repractory dermal ulcers received APG treatment for 18 times, 9 ulcers (69.2%) and 10 sinuses (88.3%) were cured at the end of 12-week treatment., Conclusion: The method of group B is the best centrifuge method. A variety of growth factors are detected and released from the platelets at significant levels in APG. There is positive correlation between the platelet concentration and PDGF-BB and TGF-beta1.

Aim: To express and characterize an active form of a single-chain antibody (scFv) from the gene of human phage antibodies which is specific for hepatocellular cancer., Methods: The complementary DNAs encoding the variable regions of the light chain (V(L)) and heavy chain (V(H)) were connected by a (Gly(4)Ser)(3) linker using a splicing by overlap extension polymerase chain reaction. The resultant scFv gene was cloned to the pET28a(+) vector and expressed in E.coli as inclusion bodies. Then the inclusion bodies were solubilized, denatured and renatured. Finally, the affinity constant of scFv was determined by noncompetitive enzyme immunoassay., Results: The target protein amounted 26% of the total protein in the condition of A(600) at 0.8 for 6 hours. After renatured, the purity of target protein was 95% and the affinity constant of the scFv was 3.6x10(7) mol/L., Conclusion: An active form of scFv which is specific for hepatocellular cancer can bind selectively with hepatocellular cancer cells, which provides a theoretical basis for immunological detection and clinical use of scFv.

This study was conducted to amplify the cfpl0-esat6 fusion gene by SOE and insert into the integrating shuttle plasmid pMV361 to form the recombinant plasmid. Then another recombinant plasmid was constructed by insertinga-A g signal sequence of BCG. The two recombinant plasmids were introduced into BCG and the induced products from recombinant BCG were analyzed. In conclusion,the successful construction of rBCG expressing the fusion protein CFP10-ESAT6 will be the base of the development of novel Mycobacterium tuberclosis vaccines.

Objective: To express a recombinant fusion protein CFP10-ESAT6 of Mycobacterium tuberculosis, and obtain the polyclonal antibodies of this fusion protein by immune rabbit., Methods: The 630 bp cfpl0-esat6 fusion gene fragments were amplified from the genomic DNA of a Mycobacterium tuberculosis reference strain H37Rv and inserted into the expression plasmid pET32a (+) to generate the recombinant plasmid pET-cfp10-esat6. The recombinat expression plasmid was transformed into E. coli BL21 (DE3). The fused protein CFP10-ESAT6 with His-tag was expressed after inducing with IPTG and purified with affinity chromatography. This protein was used to immune the rabbit to obtained the polyclonal antibodies, and been analyzed with Western-blot and ELISA., Results: The recombinant plasmid pET-cfp10-esat6 was success fully constructed, the recombinant fusion protein CFP10-ESAT6 could be expressed at relatively high levels, and the polyclonal antibodies of fusion protein were obtained., Conclusion: The successful construction and expression of the recombinant fusion protein CFP10-ESAT6 and the obtained polyclonal antibodies will be very helpful for the development of new anti-tuberculosis vaccine and the clinical serologic diagnosis.

Objective: To investigate the protective effects of recombinant human growth hormone (rhGH) on intestinal mucosal barrier in rat sepsis and explore its possible mechanisms., Methods: E. coli was injected intraperitoneally to produce rats sepsis models. Forty-two female SD rats were randomly divided into the control group (group C), sepsis group (group S) and treatment group (group T). Group S and group T were further divided into 1 d and 3 d subgroups (T1d,T3d, Sld, S3d), respectively. The expression of IGF-1 mRNA in liver, expression of Bcl-2 protein in intestine, bacteria translocation, the levels of growth hormone(GH) and insulin-like growth factor-1 (IGF-1) in plasma, and the histological appearance of intestine were determined dynamically by means of RT-PCR, radioimmunoassay, immunohistochemical staining and other corresponding methods, respectively., Results: (1) rhGH could significantly attenuate intestinal mucosal injuries and ameliorate bacteria translocation on sepsis rats. (2) The levels of Bcl-2 protein expression in intestine in group T (T1d:2441 +/- 117; T3d: 3628 +/- 235) were obviously higher than those of group S (S1d: 321 +/- 36; S3d: 1873 +/- 57) (P < 0.01). (3) The plasma levels of GH in group T (T1d: 1.28 +/- 0.24 microg/L; T3d: 2.14 +/- 0.48 microg/L) increased markedly than those of group S (S1d: 0.74 -/+ 0.12 microg/L; S3d: 0.60 +/- 0.18 microg/L) (P < 0.01). (4) The plasma levels of IGF-1 in group T (Tld: 168.94 +/- 65.67 microg/L; T3d: 201.56 +/- 64.98 microg/L) elevated significantly than those of group S (Sld: 116.72 +/- 13.96 microg/L; S3d:107.50 +/- 23.53 microg/L) (P < 0.05). (5) The levels of liver IGF-1 mRNA in group T (Tld: 0.98 +/- 0.20; T3d: 1.76 +/- 0.17) were significantly higher than those in group S (S1d: 0.38 +/- 0.09; S3d: 0.46 +/- 0.10) (P < 0.01)., Conclusion: rhGH conferred protective efficacy in maintaining the integrity of intestinal mucosal barrier against sepsis in rat. The possible mechanisms might involve the rhGH-diminished apoptosis of intestinal mucosa cells and the rhGH-maintained intestinal mucosa barrier via the roles of GH and IGF-1.

Aim: To study the induction of tumor cell apoptosis by RNA interference-mediated inhibition of the expression of telomerase in cancer cells., Methods: HeLa cells were transfected with the successfully established siRNA(small interfering RNA) expression vectors targeting hTERT (human telomerase reverse transcriptase). By electronic microscopy, Western blot and FCM (flow cytometry), the apoptosis of HeLa cells was tested., Results: The established siRNA expression vectors could induce apoptosis of HeLa cells., Conclusion: Transfection of siRNA expression vectors targeting hTERT can induce apoptosis of HeLa cells.

Objective: To investigate the molecular mechanism of quinolone-resistance of M. tuberculosis and characterize the gene mutation in Sichuan Province., Methods: Susceptibility of the clinical isolates to quinolones (ofloxacin, ciprofloxacin and sparfloxacin) was tested by the absolute concentration method. GyrA gene quinolone reasistance-determining region (QRDR) mutations M. tuberculosis were detected with PCR-SSCP and DNA sequencing., Results: Of 68 clinical isolates of M. tuberculosis, 25 high-resistant, 11 low-resistant and 10 sensitive isolates were noted to have abnormal gyrA SSCP profile and different gyrA sequences from the standard strain H37Rv, and 14 sensitive and 8 low-resistant isolates were found with no mutation of gyrA gene. DNA sequencing unveiled Ser-->Thr mutation at codon 95, Asp-->Gly at codon 94, Ala-->Val at codon 90, and Ala-->Val at codon 83., Conclusion: This study confirmed the strong correlation between the quinolone-resistance and the mutation of gyrA gene, which might be a major molecular mechanism of quinolone-resistance in M. tuberculosis. The types of mutations exhibit no difference between Sichuan Province and other areas in China.