Expert Consensus on Clinical Diseases Responding Specifically to Traditional Chinese Medicine：Fibromyalgia Syndrome / 中国实验方剂学杂志

Fibromyalgia syndrome （FMS） is a refractory， chronic non-articular rheumatic disease characterized by widespread pain throughout the body， for which there are no satisfactory therapeutic drugs or options. There are rich Chinese medical therapies， and some non-drug therapies， such as acupuncture， Tai Chi， and Ba-Duan-Jin， have shown satisfactory efficacy and safety and definite advantages of simultaneously adjusting mind and body. FMS is taken as a disease responding specifically to traditional Chinese medicine （TCM） by the National Administration of Traditional Chinese Medicine in 2018. In order to clarify the research progress in FMS and the clinical advantages of TCM/integrated Chinese and Western medicine， the China Academy of Chinese Medicine organized a seminar for nearly 20 experts in Chinese and Western medicine， including rheumatology， psychology， acupuncture and moxibustion， and encephalopathy， with the topic of difficulties in clinical diagnosis and treatment of FMS and advantages of TCM and Western medicine. The recommendations were reached on the difficulties in early diagnosis and solutions of FMS， mitigation of common non-specific symptoms， preferential analgesic therapy， TCM pathogenesis and treatment advantages， and direction of treatment with integrated Chinese and Western medicine. FMS is currently facing the triple dilemma of low early correct diagnosis， poor patient participation， and unsatisfactory benefit from pure Western medicine treatment. To solve the above problems， this paper suggests that rheumatologists should serve as the main diagnostic force of this disease， and they should improve patient participation in treatment decision-making， implement exercise therapy， and fully utilize the holistic and multidimensional features of TCM， which is effective in alleviating pain， improving mood， and decreasing adverse events. In addition， it is suggested that FMS treatment should rely on both TCM and Western medicine and adopt multidisciplinary joint treatment， which is expected to improve the standard of diagnosis and treatment of FMS in China.

Proximal femoral nail antirotation Asian version for treating femoral intertrochanteric fractures:comparison of the protruding degree of intramedullary nails in Asian population / 中国组织工程研究

BACKGROUND:Due to the mismatch between the design of the proximal femoral nail antirotation Asian version(PFNA-Ⅱ)and Asian population,extrusion of the proximal femoral intertrochanteric nail may occur in the treatment of femoral intertrochanteric fractures.The influence of the protruding length on the curative effect of the operation needs to be further discussed. OBJECTIVE:To quantitatively measure the protruding length of the proximal trochanter of the femur with intramedullary nail after PFNA-Ⅱ,and to analyze the effect of protruding length on the efficacy of PFNA-Ⅱ in the treatment of femoral intertrochanteric fractures. METHODS:Totally 68 patients with femoral intertrochanteric fractures treated with PFNA-Ⅱ internal fixation in the First Affiliated Hospital of Anhui Medical University were selected.The extramedullary process of the proximal trochanter of the femur was quantitatively measured on the anterior and posterior X-ray films of the hip joint within 6 months after operation.According to the existence of extrusion of the proximal trochanter intramedullary nail,the patients were divided into protruding group and non-protruding group.The data of sex,height,fracture type,length and diameter of the intramedullary nail,the position of screw blade in the femoral neck and protruding length of proximal greater trochanter were collected.The postoperative curative effect was judged by visual analog scale pain score and hip joint Harris score at 6 months after operation.The influence of protruding proximal trochanter of the PFNA-Ⅱ intramedullary nail on the operative effect was observed. RESULTS AND CONCLUSION:(1)There were significant differences in sexual characteristics between the protruding group and the non-protruding group(P=0.001).(2)According to AO/OTA classification,there were no significant differences in fracture type between the protruding group and the non-protruding group(P=0.289).(3)There was no significant difference in the length and diameter of the intramedullary nail between the two groups(P=0.067,P=1.000).(4)There was no significant correlation between the height of all patients and the length of the intramedullary nail(P=0.510),but there was a significant correlation between height and protruding length(P=0.034).There was no significant correlation between screw blade position and protruding length(P=0.968).(5)Six months after operation,there was no significant difference in the hip Harris score(P=0.373),but the visual analog scale pain score was significantly higher in the protruding group than that in the non-protruding group(P=0.000).(6)The results suggest that nail extrusion often occurs in the proximal greater trochanter when PFNA-Ⅱ is used in the treatment of intertrochanteric fractures in Asians.When the nail extended into the proximal soft tissue of the greater trochanter,patients complained of proximal greater trochanteric pain and the visual analog scale score of proximal greater trochanter pain in the patient was significantly higher than that in the non-protruding group.To be more suitable for the Asian population,we suggest that the PFNA-Ⅱ should be improved to further shorten the proximal nail end to obtain better clinical results of femoral intertrochanteric fracture fixation.

Correlation between abnormal thalamic functional connectivity and memory decline in patients with end-stage renal disease undergoing maintenance hemodialysis / 西安交通大学学报(医学版)

【Objective】 To explore the correlation between abnormal thalamic functional connectivity (FC) and memory loss in maintenance hemodialysis patients with end-stage renal disease (ESRD). 【Methods】 An auditory verbal learning test (AVLT-H) was conducted on 22 patients with ESRD and 28 age-, sex-, and education-matched healthy controls (HC) to evaluate memory function. After that, resting-state functional magnetic resonance imaging (rs-fMRI) data were gathered, and a whole-brain FC analysis centered on the thalamus was executed to discern variations in thalamic FC between the two groups. Finally, Pearson and Spearman correlation analyses were carried out. 【Results】 Compared to the HC group, the ESRD group exhibited notably lower scores in IR-S (P=0.002), SR-S (P<0.001), and LR-S (P=0.005). Concurrently, the ESRD group demonstrated diminished FC of the right thalamus with the left superior frontal gyrus, the left parietal lobule, the right suproccipital gyrus, the right anterior cuneus, and the right middle frontal gyrus (P<0.05, TFCE correction). Additionally, reduced FC were observed between the left thalamus and the left gyrus rectus, the left parietal lobule, and the right parietal lobule in the ESRD group (P<0.05, TFCE correction). Moreover, the FC values between the left thalamus and the left gyrus rectus in the ESRD group displayed significant negative correlations with IR-S (r=-0.499), SR-S (r=-0.458), and LR-S (r=-0.455) (all P<0.05). 【Conclusion】 Memory impairment is evident in ESRD patients undergoing maintenance hemodialysis, and it appears to be intricately linked to anomalous FC within the left thalamus and the left gyrus rectus. These findings offer potential imaging markers for monitoring memory dysfunction in individuals with ESRD.

The application value of CT in evaluation of abdominal aortic calcification in patients with chronic kidney disease / 实用放射学杂志

Objective To explore the utility of CT for the evaluation of abdominal aortic calcification(AAC)in chronic kidney disease(CKD).Methods Laboratory examination indexes and abdominal plain CT imaging of 132 CKD stage 3-5 patients were analyzed retrospectively.The ACC score was evaluated according to quantitative method,the risk factors related to AAC were analyzed.Results In correlation analyses,AAC score was positively correlated with creatinine(Cr)level,CO2 combining power(P＜0.05),and negatively corrected with albumin(ALB).Logistic regression analysis showed that Cr level were risk factors for AAC,and ALB were protective factors.Conclusion Based on abdominal plain CT images,Cr level,CO2 combining power and ALB level are related to AAC.Quantitative method based on CT has a high application value in evaluating AAC.

Effect of three-level delirium nursing management process on the treatment outcome and quality of care in the pediatric intensive care unit / 中国实用护理杂志

Objective:To explore the effect of three-level delirium nursing management process on the treatment outcome and quality of care in the pediatric intensive care unit (PICU).Methods:Since January 2019, three-level delirium management process for PICU were developed. 75 children were randomly selected as the experimental group for 6 months, and 75 patients who received routine care between July and December 2018 were selected as the experimental group. The control group received routine nursing care, and compared the differences in the incidence, duration, and length of PICU stay in the intensive care unit (DICU) between the two groups of children. The family members' satisfaction with the quality of care and the evidence-based knowledge of PICU nurses were compared before and after implementation.Results:The incidence of DICU in the experimental group was 21.33% (16/75), the incidence of unsafe events related to DICU was 2.67% (2/75), and the control group was 40.00% (30/75), 12.00% (9/75). The difference was statistically significant ( χ2 values were 6.145, 4.807, P<0.05); the duration of the DICU in the test group (14.16±4.25) hours, and the length of ICU hospitalization (48.62 ± 8.85) hours were less than (17.37±5.36) hours and (52.03±8.10) hours in the control group, the difference was statistically significant ( t values were 2.070, 2.462, P<0.05); The score of Nursing Delirium Screening Scale (Nu-DESC) for nursing care of DICU patients in the two groups was not statistically significant ( P>0.05). The score of the test group after nursing was (1.72 ± 0.75) points, which was lower than (2.17±0.63) points of the control group. The difference was statistically significant ( t value was 2.159, P<0.05); The results of the nursing quality evaluation questionnaire showed that the satisfaction of family members in the test group with respect to nursing services was 93.33% (60/75), which was higher than 82.67% (52/75) in the control group, with a significant difference ( χ2 value was 4.040, P<0.05). After the implementation, the correct answer rate of the "risk factors for delirium" questionnaire in the deliberate evidence-based knowledge questionnaire was significantly higher than before the implementation ( P < 0.05). After 6 months of implementation, the score of evidence-based knowledge of delirium was (6.82 ± 1.74) points higher than (4.26 ± 0.85) points before implementation, the difference was statistically significant ( t value was 3.739, P<0.01). Conclusions:The three-level nursing management process in the PICU ward can better prevent the occurrence of paralysis, strengthen the quality of nursing services, and exercise the comprehensive quality of nursing staff.

Unplanned tracheal extubation in the pediatric intensive care unit before and after quality management / 中国小儿急救医学

Objective To analyze the causes of unplanned tracheal extubation in mechanically ventilated patients in the pediatric intensive care unit using quality management practices and observe the changes of extubation rates before and after implementing these practices.Methods In the first place,we set up quality management group in June 2014.Second,the team formulated a risk factor checklist for unplanned tracheal extubation based on literature research, and analyzed the main causes requiring tracheal extubation. The last time,we developed some management measures for important reasons in 2014,and established and implemented a quality management strategy between 2015 and 2016.The incidence frequency of unplanned tracheal extubation,i. e. the number of extubation cases per 100 days with a tube in place(mechanical ventilation days),was used to evaluate the change of the extubation rates before and after implementing the new measures. The cases in 2014 were retrospectively analyzed,and cases from 2015 to 2016 were prospectively collected.The number of interval days for two continuity unplanned extubation was considered to be a means of quality control.Results Rick factors associated with unplanned extubation in the pediatric intensive care unit before implementing quality management were a delay of the replacing adhesive bandage, lack of sedation assessment,which were related to frequent physician replacement and insufficient training and supervision.In consideration of the identified risk factors,a standard operating procedure was formulated to prevent unplanned extubation,including standard operating procedure of tracheal catheter fixation and the sedation assessment. There were 484 mechanical ventilation in 2014,and the incidence of unplanned extubation was 0.8%,and the median number of days of separation was 8.0(4,20) d.Between 2015 and 2016,there were 1 379 mechanical ventilations,and the incidence of unplanned extubation was 0.2%,and the median number of days of separation was 34.0(19,61) d.The incidence of unplanned extubation in 2015 to 2016 was lower than that in 2014(χ2=5.936,P=0.018).Ramsay scoring usage increased from 28.6% in 2014 to 57.1% between 2015 and 2016,while Ramsay scored 3-4 points from 2015 to 2016 was higher than 2014(P< 0.05).Conclusion Establishing the quality management group according to the requirements of each hospital and implementing quality management strategies can reduce the incidence of unplanned extubation,thereby improving the quality of care in pediatric intensive care unit.

t-PA negatively regulating p75NRT for involving in atherosclerosis by changing reconstruction of vascular outer membrane autonomic nerve / 重庆医学

Objective To study the effect of tissue plasminogen activator (t-PA) on p57NTR ,inflammatory reaction ,immune regulation and oxidative stress and its effect on intimal hyperplasia .Methods The vascular injury treatment was performed in the diabetic rabbit model with carotid arterial adventitia stripping ,meanwhile t-PA controlled release microspheres were given ,the nerve distribution in the local blood vessels was observed by immunohistochemical staining .The change of nerve remodeling in the control group and treatment group was observed ,meanwhile the effect of giving t-PA controlled release microspheres on the release of ace-tylcholine and norepinephrine was detected .RT-PCR was used to detect local vascular tissue inflammation ,immune effects and oxi-dative stress .The sympathetic neurons and smooth muscle cell co-culture was adopted ,then giving glyoxal treatment as the athero-sclerosis cell model .With the t-PA treatment group as the intervention group ,the effect of t-PA on the number of cholinergic neu-ron ,and synaptic connections between the smooth muscle cells and acetylcholine secretion was observed .The change of t-PA-MMP-p75NTR and NF-kappa B signaling pathway were detected by RT-PCR .Results The vascular injury treatment was performed in the diabetic rabbit model with carotid arterial adventitia stripping ,meanwhile t-PA controlled release microspheres were given ,the nerve distribution in the local blood vessels was observed by immunohistochemical staining .The change of nerve remodeling in the control group and treatment group was observed ,meanwhile the effect of giving t-PA controlled release microspheres on the release of acetylcholine and norepinephrine was detected .RT-PCR was used to detect local vascular tissue inflammation ,immune effects and oxidative stress .The sympathetic neurons and smooth muscle cell co-culture was adopted ,then giving glyoxal treatment as the ath-erosclerosis cell model .With the t-PA treatment group as the intervention group ,the effect of t-PA on the number of cholinergic neuron ,and synaptic connections between the smooth muscle cells and acetylcholine secretion was observed .The change of t-PA-MMP-p75NTR and NF-kappa B signaling pathway were detected by RT-PCR .Conclusion t-PA activates MMPs and feedback in-hibits p75NTR-NF-kappa B signaling pathway to increase vascular adventitia autonomic nerve reconstruction and delay the occur-rence and development of atherosclerosis disease .

Surgical treatment of children sylvian cistern arachnoid cysts complicated with subdural hematoma / 中国小儿急救医学

Objective To investigate the clinical characteristics,surgical indications and methods of children sylvian cistern arachnoid cysts complicated with subdural hematoma.Methods Fifty childhood cases of sylvian cistern arachnoid cyst complicated with subdural hematoma were retrospectively analyzed,who were all surgically treated in the First Affiliated Hospital of China Medical University from July 2005 to August 2015.Among them,20 childhood cases were subacute subdural hematoma,30 childhood cases were chronic subdural hematoma.All patients underwent microscopic cyst excision,cystocistern fenestration plus hematoma removal surgery.During the surgery,firstly we resected the cyst wall tissue as far as possible,and then we communicated the cyst with subarachnoid space and cisterns,which could make the cerebrospinal fluid flowed unobstructedly.Results The course of every operation was smooth,and there was no severe complication postoperatively.Original symptoms and imaging manifestation resolved or improved in all patients.During a mean follow-up period of 5.7 years,there were no recurrent cases.Conclusion Children sylvian cistern arachnoid cyst can induce subdural hematoma.In this situation,microscopic cyst excision,cystocistern fenestration plus hematoma removal surgery is safe and effective.

Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials for repair of rabbit radius defects / 中国组织工程研究

BACKGROUND:Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were developed in our previous studies. OBJECTIVE:To observe the capability of gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials in the repair of rabbit radius defects. METHODS: Thirty healthy adult New Zealand rabbits were selected to make critical-size lacunar bone defects of the upper radius (15 mm×6 mm). Then, the rabbit models were randomized into experimental group (n=15), autogenous bone graft group (n=10) and blank group (n=5). Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were implanted into radial bone defects in the experimental group. Bone defect in blank group was implanted without any materials; in the autogenous bone graft group, the contralateral radius with same length was taken and implanted into the defect. General observation, histological observation and X-ray observation were performed respectively at 2, 4, 8, 12 weeks postoperatively. RESULTS AND CONCLUSION:At 12 weeks after operation, the experimental group showed obvious new blood vessels at the defect region, complete bony union and disappearance of the composite implant, but lamelar bone structure appeared, smal blood vessels were visible, the edge of new bone was connected to the original bone edge, exhibiting a continuity of bone, the bone density was slightly lowered, and the defect region became unobvious. In the autogenous bone graft group, bony union and trabecular bone reconstruction were distinct, the lamelar bone became mature, the medulary cavity was recanalized, the fracture line disappeared completely, and the bone density was completely consistent with that of the original bone. In the blank group, there was no obvious bone formation, which led to bone nonunion, and there were a great amount of fiber tissues and inflammatory cel infiltrated. To sum up, the gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite material can obviously promote the repair of critical-size bone defects, and the repairing effect is basicaly the same with that of autologous bone grafting.

Red blood cell lysis isolation and culture of rabbit bone marrow mesenchymal stem cells in vitro / 中国组织工程研究

BACKGROUND:Some studies have demonstrated that red blood celllysate added into the bone marrow can increase the efficiency of isolating and purifying mesenchymal stem cells, in order to obtain high-purity and high-quantity bone marrow mesenchymal stem cells. OBJECTIVE:To isolate and culture bone marrow mesenchymal stem cells of rabbit with red blood celllysis in vitro for exploring proliferation characteristics and performing the biological identification of cells. METHODS:Bone marrow suspension was col ected by puncturing the tibia with medul o-puncture needle. Red blood celllysis was added to the bone marrow suspension, and bone marrow mesenchymal stem cells were isolated and cultured. Numbers of primary cells were recorded at 4, 7, 10, and 13 days later And Growth curves of the cells at passages 2-5 were drawn for comparison of proliferative characteristics. Inverted phase contrast microscope was used to observe the morphological changes of cells. CD34, CD44 antigens of bone marrow mesenchymal stem cells were identified by flow cytometry and immunity fluorescence. RESULTS AND CONCLUSION:The adherent bone marrow mesenchymal stem cells mainly showed s spindle shape, with homogeneous nuclei, prominent nucleoli, and rich cytoplasm, which were positive for CD44 antigen and negative for CD34 antigen. The primary cells exhibited an“S”shape. Passage 4 cells had a better proliferative ability, rapider growth and more counting of cells as compared with other generations. These findings indicate that red blood celllysis method is a feasible ways of culturing bone marrow mesenchymal stem cells 0 in vitro, and passage 4 cells have the strongest proliferation capacity.

A bibliometrical analysis of academic status of Chinese core orthopaedic journals from 2005 to 2007 / 中华创伤骨科杂志

Objective To evaluate the academic status of 16 Chinese core orthopaedic journals from 2005 to 2007 in terms of general citation frequency, impact factors, allo-citation rate and specialty influence index. Methods On the basis of the normalized data published by Chinese S& T Journal Citation Reports (CJCR), the general citation frequency, impact factors, allo-citation rate and specialty influence index of the 16 Chinese core orthopaedic journals from 2005 to 2007 were ranked respectively and sta-tistically analyzed. Results In terms of the above 4 indicators, Chinese Journal of Orthopedics ranked the first, followed by Chinese Journal of Trauma and Chinese Journal of Reparative and Reconstructive Surgery. Considering its short history, our journal, Chinese Journal of Orthopedic Trauma, made a rapid progress in academic status. Conclusion Generally speaking, the 16 Chinese core orthopaedic journals enjoy a high academic quality and relatively significant impact.

Study of hypoxia-induced expression of HIF-1alpha in retina pigment epithelium.

It was shown that the expression of HIF-1alpha in retinal pigment epithelium increased under hypoxic conditions. Eight hours after the start of hypoxic exposure, the expression of HIF-1alpha reached the peak and sustained after 24-hour hypoxia. However, the morphology of PRE cells began to change and the expression of HIF-1alpha decreased after long-term (48-hour) hypoxia. Hypoxia-induced increase in the level of HIF-1alpha in RPE. It can be an important step in choriodal neovascularization.

Construction and sequencing of human interleukin-18 eukaryotic expression vector / 医学研究生学报

Objective:To construct the eukaryotic expression vector of interleukin-18 gene and carry out analysis of pVAX1-IL-18 sequence . Methods:The recombinant eukaryotic expression vector pVAX1-IL-18 was constructed by inserting interleukin-18 gene into the eukaryotic expression vector pVAX1 with molecular cloning technique . It was confirmed by restrictive enzymes(BamHⅠ/ EcoRⅠ) digestion and analysis of DNA sequencing . Similar nucleotide sequences encoding IL-18 of pVAX1-IL-18 were looked up by Blast means in NCBI GenBank. Results:Restrictive enzymes digestion analysis and DNA sequencing results revealed that the interleukin-18 gene was cloned into the eukaryotic expression vector pVAX1 successfully .The sequence of encoding IL-18 of pVAX1-IL-18 was exactly the same as the reported sequence of encoding human IL-18 in GenBank . Conclusion: The recombinant eukaryotic expression vector pVAX1-IL-18 has been constructed successfully, which lay the foundation for pVAX1-IL-18 as a genetic adjuvant of DNA vaccine.

Development of Hantavirus nucleocapsid protein for serotyping / 医学研究生学报

Hantavirus is the main cause of hemorrhagic fever with renal syndrome (HFRS). It is an acute infectious diseases characterized by fever, hemorrhage, nephritis or thrombocytopenia, and hantavirus pulmonary syndrome(HPS). The main clinical manifestations are fever, hemorrhagic lesion, acute respiratory distress and capillary leakeage syndrome. These are four different serotypes of the hantavirus species: Hantan virus(HTNV),Seoul virus(SEOV),Dobrava/Belgrade virus(DOBV),and Puumala virus(PUUV). They are known to cause HFRS, while Sin Nombre virus(SNV) causes HPS. In China, these are two serotypes of hantavirus: HTNV and SEOV found. The severity of infection depends on the viral serotype. To find a safe, rapid and specific serotyping diagnosis of the causative virus is important. The results not only can be beneficial for rodent control, but also for prevention and therapy. The current research of Hantavirus nucleocapsid protein used as serotyping antigen are summarized.

A preliminary genetic reassortment between Hantaan virus and Seoul virus strains / 中华流行病学杂志

<p><b>OBJECTIVE</b>To determine the frequency and characteristics of reassortment among Hantaan and Seoul viruses causing hemorrhagic fever with renal syndrome (HFRS).</p><p><b>METHODS</b>Mixed infections were initiated in tissue culture, using Hantaan virus strain 76 - 118 and Seoul virus strain SR-11. Potential reassortant virus plaques were picked out by multiplex RT-PCR, using primers specific for individual genome segments (L, M, S) of each strain.</p><p><b>RESULTS</b>Most of the progeny virus plaques (68.19% of 44) had parental genotype of 76 - 118 strain or SR-11 strain while 2 of 44 plaques had mixed genotypes that yielded RT-PCR bands for the same segment of both parental strains. Reassortant viruses were detected in 68.19% of 44 progeny plaques tested, involving the M and S segments. In addition, approximately 4.55% of the progeny virus plaques appeared to contain S or M segments originating from both parental virus strains, showing that they were diploid.</p><p><b>CONCLUSION</b>Genetic reassortment can occur between Hantaan virus and Seoul virus strains.</p>

The Timing of NO expression in brain tissue after diffused traumatic brain injury / 中华急诊医学杂志

Object　To determine the timing of NO expression in brain tissue after severely diffused brain injury．It would contribute to the understanding of the pathological and physiological functions of NO．Methods　On the basis of the Marmarou diffused brain injury model，the rat was killed at different hours afterward．and the NO expression in brain was measured by Griess reaction．Results　It was shown that NO in brain tissue increased quickly after trauma （5.20umol／100g），then decreased in 12h（1.96umol／100g）after　trauma，a secondary NO elevation appeared in 24h（2.31umol／100g） and 48h（2.69umol／100g） after DBI．Conclusion　It indicated that NO expression in brain tissue increased after trauma．As MCAO ischemia model，there was also NO increase at the intermediate and late stage，which might take part in the secondary pathological changes at late stage of cerebral injury．

The immune responses to hepatitis B gene vaccine in mice and the immune adjuvant effect of cytokines / 医学研究生学报

Objectives：To observe the effect of eukaryotic expression vectors coding IL-2 and IL-12 on immune responses induced by DNA immunization of HBV surface antigen（pCR3.1-S）in BABL/c(H-2d) and the protection against P815 mastocytoma cells stable expressing HBV surface antigen in mice after immunized with HBV gene vaccine.Methods：The immunization was performed by intramuscular injection,three weeks later,we directly inoculated P815-HBV-S into mice by subcutaneous injection .Tumor growth was measured every five days.Anti-HBs in serum was detected by ELISA and HBsAg specific cytotoxic T lymphocytes (CTLs) activity was measured by 51 Chromium release assay.Results：Eight weeks after immunization,the A value of mice serum in 450 nm and CTLs activity of mice codiog IL-2 and IL-12 eukaryotic expression vectors were significant higher(P<0.05) than that of mice intramuscular injected HBV-S DNA vaccine,these values are significant higher than that of mice injected pCR3.1(P<0.05).The spleen cells CTLs activity have decreased obviously after treated with anti-CD8+ monoclonal antibody and have no significant change after treated with anti-CD4+ monoclonal antibody.The HBV-S gene vaccine could evidently inhibit the tumor growth,prolong the survival period (>38.2 days) and improve the survival rate in mice.Conclusions：The DNA vaccine of HBV ( pCR3.1-S) had strong antigenicity in cellular and humoral immunity and had marked killing effect on HBV infected cells in vivo，which could be promoted by vector coding murine IL-2 or IL-12.CTLs activity was performed by CD8+ cells.

Raised immune responses of HBV preS_2S DNA vaccines by administrating genetic adjuvant at different time / 中国免疫学杂志

Objective:To study the different effect of fused gene IL-2/Fc administrated at different time on immune responses induced by HBV preS_2S genetic vaccine.Methods:BALB/c mice were immunized with purified recombinant plasmids at 0, 2, 4 weeks. Two groups were designed. One group were injected with pcDNA3.1S_2S at first, then pcDNA3.1IL-2/Fc was administrated after three days. While the other group were injected two plasmids at the same time. We compared the immune responses through detecting the anti-HBs titers, CTL cytotoxicity level, proliferation of lymphocytes and cytokines levels secreted by cultural splenocytes.Results:The data demonstrated that the humoral and cellular immune responses induced by pcDNA3.1IL-2/Fc as adjuvant and administrated after HBV preS_2S DNA vaccine 3 days mice were dominant higher than other groups.Conclusion:Genetic adjuvant administrated after Ag priming can enhance its effect.

Construction and expression of eukaryotic expression vector harboring human neutrophil peptide 1 / 解放军医学杂志

Objective To explore the possibility of human neutrophil peptide 1 (HNP1) gene engineering, we construct the eukaryotic expression vector carrying HNP1 gene. Methods With RNA extracted from human neutrophil cell as template, cDNA encoding mature HNP1 was amplified by RT-PCR, and then it was inserted into vector pMD18-T. After restriction endonuclease digestion and DNA sequencing confirmation the gene was subcloned into eukaryotic expression plasmid pcDNA3.1/V5-His-TOPO to construct a recombinant expression plasmid pcDNA3.1/V5-His-TOPO/HNP1, then the recombinant plasmid was transfected into COS-7 cells by lipofectamine, and the expressed product was identified by biotin-avidin enzyme-linked immunosorbent assay ( BA-ELISA). Results The sequence of HNP1 completely matched those published in GenBank, thus eukaryotic expression vector pcDNA3.1/V5-His-TOPO/HNP1 was constructed correctly. The ELISA results showed that the eukaryotic expression plasmid pcDNA3.1/V5-His-TOPO/HNP1 could temporarily express HNP1 in COS-7 cells. Conclusion The successful construction and expression of pcDNA3.1/V5-His-TOPO/HNP1 pave the way for the stable expression HNP1 in mammalian engineering cells.

Inhibition of VEGF gene expression in Hela cell by short interfering RNAs / 中国免疫学杂志

Objective:To investigate the interfering effect of siRNA on the expression of VEGF in Hela cells.Methods:Three pairs of siRNA were designed according to the 1-5 extrons sequence of VEGF and synthesized by T7 RNA ploymerase transcription in vitro. To evaluate the inhibition activity of siRNA, Hela cells were transfected via siPORT Lipid. The interfering effect of siRNAs in hRPE cells was examined by semi-quantitative RT-PCR and immunofluorescence technique.Results:The results showed that the three pairs of siRNA could effectively inhibit gene expression of VEGF in Hela cells with the rates of 88.7%, 94.2% and 80.3%. But the 1-base mismatched siRNA and ssRNA(+) didn′t show significant interfering effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect(5-10 pmol).Conclusion:The siRNAs synthesized by T7 RNA ploymerase in vitro could effectively and specifically interfere the expression of VEGF in Hela cells, providing a novel approach for gene therapy of tumor.