An eukaryotic expression plasmid Rc/CMV GHcDNA containing CMV late promoter and hGH cDNA was constructed and introduced into mouse skeletal muscle by means of direct injection with or without the cationic liposome (Lipofectin and Lipofectamine reagent) intramuscularly. The expression of the gene was detected through both RT-PCR and IRMA (Immunoradiometric assay) at the level of transcription and protein respectively. The gene expression can be detected even after 90 days of single plasmid injection. Compared with the mice injected with the plasmid itself, the expression levels of mice injected with the plasmid-cationic liposome complex seem to be higher, and the expression period longer. Furthermore, the effect of Lipofectamine seems to be even better than that of Lipofectin. The results indicate that direct intramuscular injection with recombinant expression plasmid or plasmid-cationic liposome complex is a simple, efficient and economical way for foreign gene transfer and expression in experimental mice in vivo.

Objective To analyze the effectiveness of antithymocyte globulin(ATG) combined with cyclosporin(CSA)in treatment of pediatric severe aplastic anemia(SAA).Methods Seven children with SAA were treated with ATG and CSA,ATG 4-5 mg/(kg?d),5 days,methyprednisone 2-3 mg/(kg?d) to reduce anaphylaxis,CSA 3-5 mg/(kg?d),assisting with transfusion and G-CSF.Results The complete remission rate was 28.57%(2/7),the partial remission rate was 14.29%(1/7),and the obvious improvement rate was 28.57%(2/7),1 child died.The overall response rate was 71.43%.Transfusion volumes in 4 to 6 months post treatment decreased obviously comparing to the first 3 months(P

OBJECTIVE: To evaluate the efficacy and safety of proximal femoral nail antirotation(PFNA) vs InterTAN nail in treating the elderly intertrochanteric femoral fractures. METHODS: Data of the randomized controlled trials(RCTs) about PFNA vs InterTAN for the treatment of the elderly intertrochanteric femoral fractures were searched in as the Cochrane Library, PubMed, EMbase, Wanfang, CNKI, CBM and VIP from their establishment to January 2018 for collecting. After study selection, assessment and data extraction conducted by two reviewers independently, meta-analyses were performed by using the RevMan 5.3 sofware. The level of evidence was assessed by using the GRADEpro system. RESULTS: Twelve studies involving 1 015 patients were included. The results of meta, analyses showed that: (1)safety indicator: compared with the InterTAN, PFNA had shorter operation time, and less intraoperative bleeding. But InterTAN had less total postoperative complications and internal fixation failure, but there was no significant difference in the operative incision lengths, fracture healing time and other postoperative complications. (2)efficacy indicator: compared with the InterTAN, the Harris hip score was lower after 3 months, but Harris hip score had no significant difference between the two groups after 6, 12 months. Based on GRADEpro system, all the evidence was at level C and weak recommendation(2C). CONCLUSIONS The current evidence indicates that PFNA had a similar effect compared with the InterTAN. But InterTAN could provide better stability against rotation and axial pressure effect, can allow patients do functional exercise early such as ambulation to recovery the hip function. It also had less total postoperative complications and internal fixation failure for the poor quality of the original studies and the limited number of studies, a prudent choice is suggested and more high-quality, large-sample studies are need.
Aged ; Bone Nails ; Femoral Fractures ; surgery ; Fracture Fixation, Internal ; Fracture Fixation, Intramedullary ; Humans ; Treatment Outcome

Objective To explore the possibility of three-dimensional (3D) kidney replications for clinical and teaching of percutaneous nephrolithotomy (PCNL).Methods The CT urography (CTU) DICOM format data from 5 patients with kidney calculi were selected from March 1st to June 1st,2015.Thresholding technique,region growing technique,edit mask technique and multiple slice edit technique were used in sequence by Mimics software.And five 3D replications were printed by Object 500 3D printer.The sizes of the replications were measured and the replications were punctured.Results The 3D replications of the kidneys were successfully designed and printed.The average difference of the long axes between 3D replications and patients' kidneys was 0.283cm,the average difference of the diameters was 0.212cm,and the average difference of the diameter of the stones was 0.244cm.The sizes of the 3D replications were basically consistent with those of patients' kidneys.The simulative puncturing was successful.Conclusions After comparing the 3D replications with their original 2D CT images,the anatomical details are found basically the same.The 3D replications could provide 3D visual observations of organ anatomy for surgeons.

In this study, we designed and synthesized 12 novel aloperine derivatives with different core structures. Among them, compound 3 with a ten-membered ring core was obtained through a special ring expansion reaction after γ-H Huffman elimination of quaternary ammonium salt, and the structure was verified by X-single crystal diffraction. Furthermore, their antiviral activity against human β-coronavirus HCoV-OC43 was evaluated by CCK-8 assay. Quaternary ammonium salt 2a and 3 had a good inhibitory effect against HCoV-OC43, and 2a had the highest anti-HCoV-OC43 activity with an EC₅₀ values of 3.77 μmol·L^-1 and a SI value of over 53.1. Schrӧdinger molecular docking results showed that both 2a and 3 might display their anti-HCoV-OC43 activity by directly acting on host TMPRSS2 and SR-B1. The results expanded the structural types of endocyclic aloperine and the function against coronavirus, and provided useful scientific data for the development of pharmaceutical applications of these compounds.

Abstract: Objective To detect the polymorphisms of drug resistance-related genes pvcrt-o and pvmdr1 of Plasmodium vivax in lazan city in the China-Myanmar border, in order to guide the treatment plan of Plasmodium vivax. Methods A total of 48 Plasmodium vivax samples were collected from Lazan in the China-Myanmar border in 2007, and fragments of pvcrt-o and pvmdr1 genes were amplified by PCR and sequenced. The sequences were aligned with the Salvador I (Sal-I) strain reference genome sequences to determine the presence of SNPs. Results The target fragments of pvcrt-o gene were amplified from 39 Plasmodium vivax samples, while pvmdr1 genes were amplified from 40 samples. Amongst them, 25 samples had AAG insertion before the 10th amino acid (K10 insertion) of pvcrt-o gene, accounting for 64.1%. Non-synonymous mutations were detected at three loci of pvmdr1 gene (T958M, Y976F, and F1076L), the mutation rates were 100%, 22.5%, and 55.0%, respectively. There were three haplotypes of pvmdr1 gene, of which the triple mutant 958M/976F/1076L accounted for 22.5% (9/40), the double mutant 958M/Y976/1076L accounted for 32.5% (13/40), and the single mutant 958M/Y976/F1076 accounted for 45.0% (18/40). The proportion of strains with pvcrt-o and pvmdr1 gene mutation is 63.16%, which is significantly different from those only with pvmdr1 mutation. Conclusions The proportion of pvcrt-o and pvmdr1 gene mutation of 48 Plasmodium vivax isolates is high in the China-Myanmar border, and there is a certain degree of correlation between the two gene mutations. To assess changes in Plasmodium vivax drug resistance in this region, it is required to improve the surveillance of these two molecular markers.

Objective: To investigate the sonographic features of primary thyroid lymphoma (PTL) and to evaluate the clinical significance of ultrasound-guided core needle biopsy (US-CNB) in PTL. Methods: A total of 24 patients with suspected PTL in Sir Run Run Shaw Hospital from January 2013 to June 2018 were analyzed retrospectively. All cases were confirmed by pathology, of them 23 patients received US-CNB and 1 patient chose operation without US-CNB, including 5 males and 19 females, aged from 39 to 75 years old. The effectiveness and safety of 23 patients with US-CNB were evaluated, and the sonographic features of 20 patients with PTL diagnosed by pathology were analyzed. Descriptive statistical methods were used in the study. Results: In the 23 patients with suspected PTL underwent US-CNB, 18 patients were diagnosed as PTL, 4 patients were respectively diagnosed as subacute thyroiditis, anaplastic carcinoma, Hashimoto's thyroiditis, and fibro thyroiditis, and the another patient was hard to diagnose by US-CNB and then was diagnosed as PTL by surgical biopsy. The success rate of US-CNB for diagnosis of PTL was 18/19, and no severe complications occurred in the patients with US-CNB. The other case was diagnosed as PTL by surgical biopsy without US-CNB. Sonographic features of 20 cases with PTL (18 cases diagnosed by US-CNB and 2 cases by surgery or surgery biopsy) were as follows: (1) Most nodules had irregular shapes and unsmooth margins; (2) Hypoechoic or markedly hypoechoic nodules with honeycombed or cord structures were observed in most cases; (3) Calcification was rare; (4) Multiple lesions were common; (5) Abundant intralesional vascularization was commonly observed; (6) Most cases had intensification of posterior acoustic enhancement; (7) Thyroid gland enlargement or with irregular shape; and (8) PTL often accompanied with lymph nodes enlargement in lateral neck or central region. Conclusion: PTL has certain sonographic features, with assistance of US-CNB, more accurate diagnosis of PTL can be obtained.
Adult ; Aged ; Biopsy, Large-Core Needle ; Female ; Humans ; Image-Guided Biopsy ; Lymphoma/diagnostic imaging* ; Male ; Middle Aged ; Retrospective Studies ; Thyroid Gland ; Thyroid Neoplasms/diagnostic imaging* ; Thyroid Nodule ; Ultrasonography, Interventional

OBJECTIVETo determine the enhancement effects of caffeine on chemotherapy of transplanted osteosarcoma in Fischer 344/N rats.

METHODSOsteosarcoma-bearing Fischer 344/N rats were treated with cisplatin 2.5 mg/kg (Group DDP), caffeine 90 mg/kg x 2 d (Group caffeine), and cisplatin 2.5 mg/kg plus caffeine 90 mg/kg x 2 d (Group DDP+caffeine), and the control group was treated with normal saline in the same volume. All drugs were given by intra-peritoneum injection with micro-pump, in the rate of 0.5 ml/h. The tumor volume was measured and evaluated. The tumors were stained in TUNEL, and PCNA was detected with immunohistochemistry. The tumor growth inhibition rate, PCNA index and apoptosis index were calculated, and the survival time were recorded.

RESULTSThe tumor inhibition rate was -0.5219 +/-0.1429 in control group, 0.0362 +/-0.0957 in Group DDP, -0.4193 +/-0.1345 in Group caffeine, and 0.3646 +/-0.1313 in Group DDP+caffeine (P <0.01). PCNA index was 0.4587 +/-0.1312 in control group, 0.1847+/-0.0535 in Group DDP, 0.4381 +/-0.0706 in Group caffeine, and 0.0314 +/-0.0231 in Group DDP+caffeine (P <0.01). Apoptosis index was 0.0008 +/-0.0005 in control group, 0.0077 +/-0.0060 in Group DDP, 0.0011 +/-0.0003 in Group caffeine, and 0.0295 +/-0.0069 in Group DDP+caffeine (P <0.01). And the survival time was (33.63 +/-4.63)d in control group, (52.13 +/-11.74)d in Group DDP, (35.63 +/-5.15)d in Group caffeine, and (55.13 +/-16.23)d in Group DDP+caffeine (P <0.01).

CONCLUSIONCaffeine could enhance the anti-tumor effect of cisplatin in rat osteosarcoma.

Animals ; Antineoplastic Agents ; therapeutic use ; Bone Neoplasms ; drug therapy ; pathology ; Caffeine ; therapeutic use ; Cisplatin ; therapeutic use ; Drug Synergism ; Neoplasm Transplantation ; Osteosarcoma ; drug therapy ; pathology ; Rats ; Rats, Inbred F344

BACKGROUNDErythropoietin (EPO) functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production and neuron protection. This study aimed to determine the neuron protective effect of erythropoietin on experimental rats enduring spinal cord injury (SCI) by assessing thrombospondin-1 (TSP-1) level and transforming growth factor-beta (TGF-beta) in the development of a rat model of SCI.

METHODSSixty Sprague-Dawley rats were randomly assigned to three groups: sham operation control group, SCI group and EPO treatment group. By using a weight-drop contusion SCI model, the rats in the SCI group and EPO treatment group were sacrificed at 24 hours and 7 days subsequently. The Basso, Beattie, and Bresnahan (BBB) scores were examined for locomotor function. Pathological changes were observed after HE staining. The expressions of thrombospondin-2 (TSP-1) and TGF-beta were determined by immunohistochemical staining and Western blotting.

RESULTSSlighter locomotor dysfunction was discovered and it was recovered abruptly as higher BBB scores were found in the EPO treatment group than in the SCI group (P < 0.01). Pathologically, progressive disruption of the dorsal white matter and regeneration of a few neurons were also observed in SCI rats. TSP-1 and TGF-beta expression increased at 24 hours and 7 days after SCI in the injured segment, and it was higher in the SCI group than in the EPO treatment group. Spinal cord samples from the animals demonstrated a TSP-1 optical density of 112.2 +/- 6.8 and TSP-1 positive cells of 5.7 +/- 1.3 respectively. After injury, the TSP-1 optical density and cell number increased to 287.2 +/- 14.3/mm(2) and 23.2 +/- 2.6/mm(2) at 24 hours and to 232.1 +/- 13.2/mm(2) and 15.2 +/- 2.3/mm(2) at 7 days respectively. When EPO treated rats compared with the SCI rats, the TSP-1 optical density and cell number decreased to 213.1 +/- 11.6/mm(2) and 11.9 +/- 1.6/mm(2) at 24 hours and to 189.9 +/- 10.5/mm(2) and 9.3 +/- 1.5/mm(2) at 7 days, respectively (P < 0.01). In the SCI rats, the TGF-beta optical density and positive neuron number were 291.4 +/- 15.2/mm(2) and 28.8 +/- 4.9/mm(2) at 24 hours and 259.1 +/- 12.3/mm(2) and 23.9 +/- 4.1/mm(2) at 7 days respectively. They decreased in the EPO treated rats to 222.8 +/- 11.9/mm(2) and 13.7 +/- 2.1/mm(2) at 24 hours and to 196.5 +/- 9.7/mm(2) and 8.7 +/- 2.2/mm(2) at 7 days (P < 0.01).

CONCLUSIONSIncreased expression of TSP-1 and TGF-beta can be found in the injured segment of the spinal cord at 24 hours and 7 days after injury. EPO treatment can effectively prevent pathological alterations from severe spinal cord injury by reduced expression of TSP-1 and TGF-beta.

Animals ; Blotting, Western ; Disease Models, Animal ; Erythropoietin ; therapeutic use ; Female ; Immunohistochemistry ; Neuroprotective Agents ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; drug therapy ; metabolism ; Thrombospondin 1 ; metabolism ; Transforming Growth Factor beta ; metabolism

Adult ; Gallbladder Diseases ; pathology ; Gastric Mucosa ; pathology ; Humans ; Male