The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Background: Ribosomal proteins are traditionally associated with protein biosynthesis until recent studies that implicated their extraribosomal functions in human diseases and cancers. Our previous studies using GeneFishingTM DEG method and microarray revealed underexpression of three ribosomal protein genes, RPS26, RPS27, and RPL32 in cancer of the nasopharynx. Herein, we investigated the expression pattern and nucleotide sequence integrity of these genes in nasopharyngeal carcinoma to further delineate their involvement in tumourigenesis. The relationship of expression level with clinicopathologic factors was also statistically studied. Methods: Quantitative Polymerase Chain Reaction was performed on nasopharyngeal carcinoma and their paired normal tissues. Expression and sequence of these three genes were analysed. Results: All three ribosomal protein genes showed no significant difference in transcript expressions and no association could be established with clinicopathologic factors studied. No nucleotide aberrancy was detected in the coding regions of these genes. Conclusion: There is no early evidence to substantiate possible involvement of RPS26, RPS27, and RPL32 genes in NPC tumourigenesis.

Purpose: A phase II study was conducted to evaluate the safety and efficacy of preoperative, intra-arterial perfusion of epirubicin, etoposide, and oxaliplatin combined with oral chemotherapy S-1 (SEEOX) for the treatment of type 4 gastric cancer. Materials and Methods: A single-center, single-arm phase II trial was conducted on 36 patients with histologically proven type 4 gastric cancer without distant peritoneal or organ metastasis. Patients received 3, 21-day courses of SEEOX preoperative chemotherapy. The primary endpoint was overall survival (OS) and the secondary outcomes assessed were chemotherapeutic response, radical resection rate, pathological regression, toxicities, postoperative morbidity, and mortality. Results: All patients were at an advanced stage of cancer (stage III or IV) and completed the entire course of treatment. Based on changes in tumor volume and peritoneal metastasis, the objective response rate was 55.6% (20/36; 95% confidence interval [CI], 38.5%–72.6%) and the disease control rate was 69.4% (25/36; 95% CI, 53.6%–85.3%). The radical resection rate was 75% (27/36; 95% CI, 60.1%–89.9%) and the proportion of R0 resections was 66.7% (21/36; 95% CI, 50.5%–82.8%). The pathological response rate was 33.3%, of which 13.9% showed complete pathological regression. The median survival was 27.1 months (95% CI, 22.24–31.97 months), and the 2-year OS was 48.5% (95% CI, 30.86%–66.1%). Conclusions Preoperative SEEOX is a safe and effective treatment for type 4 gastric cancer. Based on these preliminary data, a phase III study will be conducted to confirm the superiority of this regimen over standard treatment.Trial Registration: ClinicalTrials.gov Identifier: NCT02949258

Objective To establish a kind of animal model and methods for neuroendoscope training. Methods With rat abdominal cavity hypothesized as cerebral ventricle, a set of programs for neuroendoscopic operative skill training were designed, including endoscopic techiniques, electronic coagulation, suction, flush, biopsy and balloon dilatation, and so on. Results Simulation operation was performed on rat abdomen under neuroendoscope. The procedure helped to practice the endoscopic manipulation, get to know well how to perform endoscope, coagulation, suction, flash instruments, be familiar with the usage of electric coagulation in the liquid condition. The emphasis was put on the basic skills of pure neuroendoscopic operation such as balloon dilatation, electric coagulation, cutting off,forcep biopsy and flush. Conclusions The rat model can provide a way to train pure neuroendoscopic operation. The trainer can get basic experience with regard to endoscopic manipulation, balloon dilatation, electric coagulation, suction, flush and biopsy under endoscope. The method can be an important part of neuroendoscopic laboratory training.

Objective To evaluate the influence and significance of preoperative design of surgical approach on stem cell transplantation via stereotactic surgery. Methods Six patients with stroke in the basal ganglia region were selected. The transplantation target and transcranial approach point were designed by magnetic resonance examination before stem cell transplantation via stereotacfic surgery to guarantee that the line connecting the transplantation target and transcranial approach point could avoid the important functional areas, the ventricular system and the softening focus. Postoperative magnetic resonance examination was performed to observe whether the practical target and surgical approach coincided with the preoperative design or not. Results The practical transplantation target was coincided with the designed transplantation target, distributed around the softening focus without implanted cells in the softening focus. Surgical approach was coincided with the preoperative design and it successfully avoided the important brain functional area, ventricular system and softening focus.Conelnsion The preoperative design of surgical approach can not only ensure the cells being exactly transplanted into the reservation target and guarantee the curative effect, but also promise the surgical approach successfully avoiding the important brain functional area, ventricular system and softening focus and reduce the operative injury.

OBJECTIVETo observe the effects of Ganoderma lucidum spores on Cytochrome C (Cyt-C) and mitochondrial calcium in the testis of NIDDM rats.

METHODSFifty male Wistar rats were divided randomly into three groups: model, ganoderma and normal control, the first two groups injected with 2% STZ through vena caudalis, and the last one with half-and-half sodium citrate/citrate buffer solution. Two weeks after normal diet, glucose tolerance tests were performed and the rats with abnormal glucose tolerance from the model and ganoderma groups received high-fat and high-carbohydrate food, the ganoderma group given Ganoderma lucidum spores (250mg/[ kg x d] ) in addition, both for 10 weeks. Glucose tolerance tests were repeated 1 day before the end of the experiment and the rats were castrated and relevant indexes measured.

RESULTSThe NIDDM model was successfully constructed. In the model group, the levels of mitochondrial Cyt-C and mitochondrial calcium were significantly lower (P <0. 05) while that of the plasma Cyt-C was significantly higher than in the ganoderma and the control groups.

CONCLUSIONCyt-C and calcium ion are involved in the damage of the testis. Ganoderma lucidum spores can protect the testis of NIDDM rats.

Animals ; Calcium ; metabolism ; Cytochromes c ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Male ; Mitochondria ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Reishi ; Testis ; drug effects ; metabolism

[Objective]To apply the isobaric tagging for relative and absolute quantitation (iTRAQ) and liquid chromatography-tandem mass spectromeury (LC-MS/MS) to identify the proteins differentially regulated in CSF of patients before and after stem cell transplantation.[Methods]CSF samples fiom patients with central nervous system injury before and after stem cell transplantation and from normal controls were collected;selective removal of high-abundant proteins was performed by high performance liquid chromatography;proteins were differentially identified by iTRAQ-coupled LC-MS/MS.[Results] Totally,59 proteins were identified.As compared with control group,the patients before transplantation had 5 over-expressed proteins and 8 under-expressed proteins;only 7 proteins in patients after transplantation showed no significantly different expression as compared with those in control group.[Conclusion] Protein expression is significantly changed in CSF of patients with central nervous system injury after neural stem cell transplantation,suggesting that the proteomics research can help us to find markers of nervous system damage and is important to mechanism study on nervous system damage and repair.

Objective To analyze and study the prescriptions of Chief Physician Chang Yalin for treating breast hyperplasia patients in the outpatient department,explores and summarizes the medication patterns for treating breast hyperplasia,and provides reference and dialectical medication ideas for clinical treatment of breast hyperplasia by using data mining technology.Methods Collect the prescriptions of Dr.Chang Yalin for treating breast hyperplasia patients at the Outpatient Department of Jiuquan Traditional Chinese Medicine Hospital from January 1,2017 to December 31,2022.After induction and organization,establish a data table,use Excel 2019 to statistically analyze the drug frequency and gender,taste,and meridian.Use SPSS Modoler 18.0 software April algorithm to analyze the association rules of the drugs,and use SPSS Statistics for cluster analysis.Results A total of 184 prescriptions were included,covering 140 types of traditional Chinese medicine,with a total frequency of use of 4059 times.Among them,Cornus officinalis,dried ginger,roasted licorice,Poria cocos,and jujube are commonly used drugs for the treatment of breast hyperplasia.The top three types of Chinese medicine that are commonly used are tonifying deficiency,warming internal medicine,and relieving external symptoms.The medicinal properties are mainly warm and mild;The medicinal taste is characterized by sweetness,bitterness,and bitterness;The main meridians of drugs are spleen,lung,and heart meridians.Association rule analysis shows that roasted licorice white peony,poria cocos,licorice,and cornus flesh white peony,jujube,and ginger are equivalent.Classify commonly used drugs into 4 categories using Euclidean distance clustering and segmentation with 5.Conclusion Dr.Chang Yalin has always implemented the core idea of treating breast hyperplasia through the coordination of liver and spleen,the simultaneous application of attack and tonifying,and the simultaneous treatment of phlegm and blood stasis.In terms of the combination of latent drugs,the main focus is on tonifying qi and spleen,tonifying yin/blood and regulating liver,and resolving stasis/resolving phlegm and dispersing stagnation.In terms of the combination of"cold phlegm"and"blood stasis and toxin",as well as the combination of"eighteen contraindications"and"nineteen contraindications",his principles and methods are more distinctive and worthy of further research and promotion.

The purpose of this study was to investigate the epidemiological characteristics and risk factors of influenza in Hainan province,to provide evidence to support influenza prevention and control efforts.Pathogen monitoring data of influenza-like illness(ILI)in six national sentinel hospitals in Hainan province from 2013 to 2021 were analyzed in SPSS 20.0 software.A total of 50 415 ILI cases were detected during the 2013-2021 season,of which 5 581 were positive for influenza viruses,with a positivity rate of 11.07％.The dominant strains were type B,type A(H1N1)pdm09 and type A(H3N2).The positivi-ty rate of influenza virus was highest in people 5-14 years of age(17.56％)and lowest in people 0-4 years of age(7.32％).Influenza activity showed both a summer peak and a winter-spring peak in the 2014-2016,2017-2018 and 2019-2020 sea-sons,and was concentrated in April to September,with a maximum peak of 53.64％,and in November to March of the next year,with a peak of 47.30％.The 2013-2014,2016-2017 and 2018-2019 seasons showed only a winter-spring peak concen-trated between October and March of the next year,with a maximum peak of 54.17％,but no obvious summer peak.The pre-dominant influenza viruses during the eight surveillance seasons varied among H1N1,H3N2 and type B.The positive detection rate of influenza virus steeply declined during the 2020-2021 season:the positive detection rate was only 0.25％,and no obvi-ous epidemic period was observed.The intensity of influenza epidemic varied among monitoring years,and the dominant strains changed rapidly in Hainan Province.People 5-14 years of age were the key population affected.Summer,winter and spring were the key periods for influenza prevention and control.Etiological surveillance of influenza should continue to be strength-ened,the roles of health education and publicity should be emphasized,and the dual measures of influenza vaccination and non-drug intervention should be actively promoted to decrease the occurrence of influenza.

Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies.