To study the method and the optimal quantity of osteoblast transplantation to stimulate bone regeneration by transplanting 6～8 day old cultured osteoblasts and bFGF, together with osteoblast free calvaria of fetal rabbits into the defects of the rabbit radius. The fibroblasts were embedded in the type Ⅰcollagen in the concentrations of 10 7 ,10 6 ,10 5 , 10 4 /ml. Bone repair was evaluated by X ray, bone density,and scanning electron microscopy. Bone union ratio of the radius with transplantation of 10 7 ,10 6 ,10 5 , 10 4 /ml osteoblasts was 40%, 65%, 31 25%, 5 55% respectively at 4 weeks after transplantation, and 41 67%, 75%, 37 5%, 10% respectively at 8 weeks after transplantation. Bone density of radius defect with transplantation of 10 7 , 10 6 , 10 5 , 10 4 /ml osteoblasts was (0 112?0 018), (0 159?0 033), (0 122?0 039), (0 066?0 002) respectively at 6 weeks after transplantation and (0 150?0 059), (0 173?0 041), (0 145?0 023), (0 103?0 023) respectively at 8 weeks after transplantation, In 10 cases of bone non union, union was achieved by transplanting fetal osteoblast and basic fibroblast growth factor. This finding indicated that that transplantation of fetal osteoblasts could stimulate bone defect repair. The optimal concentration of osteoblasts implant was 10 6 cells /ml. It might be used in the treatment of bone non union.

Objective A design of guide wire localization apparatus for the internal fixation of dens fractures. Methods From October 1999 to February 2000, 5 cases of dens fractures were treated by anterior internal fixation using cannulated compression screws; 4 cases were of Anderson typeⅡ and 1 case was typeⅢ . The guide wire localization apparatus was used for all of the patients during operations. Results The operating time was 2 to 2.5 hours, averaging 2 hours 15 minutes. The amount of bleeding was 20 to 100 ml, averaging 60 ml. The time of follow-up was 5 to 9 months, averaging 6.7 months. The locations of all the 5 cannulated compression screws were excellent, positioning in the center of the odontoid process without deviation. 4 cases healed normally and 1 had delayed bone healing. Conclusion The use of a guide wire location apparatus increased the accuracy of internal fixation of screws in dens fracture, decreased X-ray to radiation to the bodies, and reduced the possibility of pollution in operation.

OBJECTIVE: To explore the therapeutic effect of tetrandrine(TET) on silicosis model rats and its toxic effect on liver and kidney function. METHODS: The specific pathogen free healthy male Wistar rats were randomly divided into the control group, the model group and the TET group, with 14 rats in each group. By un-exposure tracheal injection method, the rats in the model and TET groups were given one-time tracheal infusion of free silicon dioxide suspension with a mass concentration of 50 g/L to establish the rat model of silicosis. Rats in the control group were infused with 1 mL of 0.9% sodium chloride solution with the same method. On the second day after the model was established, the TET group was given 30 mg/kg body mass of TET solution by gavage. The other two groups were given the same amount of 0.9% sodium chloride solution. The treatment was once per day, six times per week. Seven rats in each group were sacrificed on the 28 th and 56 th days after modeling. The morphological change of the lung, liver and kidney tissues of each group was observed. The enzyme-linked immunosorbent assay was used to detect the level of tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1), interleukin(IL)-1β and IL-6, in the lung tissues of rats in each group. The activities of aminotransferase(ALT), aspartate aminotransferase(AST) and the levels blood urea nitrogen(BUN), creatinine(CRE) were detected by automatic biochemical analyzer. RESULTS: The lung organ coefficients of rats in the TET group were lower than those of the model group on the 28 th and 56 th days(all P<0.05). The lung organ coefficient of the rats in the TET group on the 56 th day was higher than that in the same group on the 28 th day(P<0.05). The lung tissue structure of the control group was normal. After modeling, the lung tissues of rats in model group showed different degrees of pathological changes such as alveolar structure destruction, inflammatory cell infiltration, and fibrosis on the 28 th and 56 th days. The degree of pathological changes in TET group was less than that of the model group. In the lung tissues of rats in the model group, the levels of TNF-α, TGF-β1, IL-1β and IL-6 were higher than those of the control group(all P<0.01). The levels of TNF-α, TGF-β1, IL-1β and IL-6 in the lung tissues of rats in the TET group were lower than that of the model group(all P<0.01), but there was no statistically significant difference when compared with the control group(all P>0.05). The activities of ALT and AST in the TET group were higher than those in the model group and the control group(all P<0.01). The level of serum BUN in TET group was higher than that in control group(P<0.01), but it showed no statistical difference when compared with the control group(P>0.05). The level of serum CRE in each group showed no significant difference(P>0.05). There were no abnormal pathological changes found in the liver and kidney tissues of rats in each group at different times. CONCLUSION: TET can reduce the inflammatory response in silicosis rats and improve lung tissue fibrosis; however, the therapeutic dose may have certain toxicity to the liver and kidney of the silicosis rats.

OBJECTIVETo investigate the effects and mechanisms of Yishen Huoxue decoction (YHD) on chronic renal failure (CRF) rats induced by 5/6 nephrectomy.

METHODSThe glomerulosclerosis model was established by 5/6 nephrectomy in rats. Experimental animals were allocated into the normal group, the model group, the YHD group and the benazepril group. Urine protein of 24 h (UP) at the 6th and 12th weekend after operation, blood urea nitrogen (BUN) and creatinine (SCr), albumin (Alb) and haemoglobin (HB) at the 12th weekend were measured, renal pathology changes were examined with light microscope, the expressions of proliferating cell nuclear antigen (PCNA) and fibronectin (Fn) were examined by immunohistochemistry and mRNA expressions of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) by RT-PCR at the 12th weekend.

RESULTSCompared with those in the normal group, the levels of UP, BUN and SCr, the area of glomerular mesangial matrix, the FN deposition, PCNA expression in glomeruli and tubular interstitium and mRNA expressions of CTGF and PAI-1 were all significantly higher in the model group (P < 0.05). All the above-mentioned indexes were lower in the YHD group than those in the model group (P < 0.05). PCNA positively expressed cells in glomeruli of the normal, model group, YHD group and benazepril group was 7.00 +/- 2.24,34.78 +/- 6.96,15.75 +/- 2.61 and 15.50 +/- 2.57 respectively, positively correlated to the expression of CTGF, PAI-1, FN and SCr level.

CONCLUSIONYHD could delay the progression of CRF in 5/6 nephrectomized rats, and the mechanisms were mainly related to the inhibition on renal cell proliferation and it induced over-expression of cytokines, and accumulation of extracellular matrix.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; biosynthesis ; genetics ; Glomerulosclerosis, Focal Segmental ; drug therapy ; etiology ; metabolism ; Kidney Failure, Chronic ; drug therapy ; etiology ; metabolism ; Male ; Nephrectomy ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

We studied the construction of fusion protein TAT-RIG-I-GFP prokaryotic expression vector and verified the function of TAT in transmembrane delivery. First, four pairs of specific primers were designed, and the RIG-I gene of Mallard Duck (Anas platyrhynchos) was cloned. Then, the pET-TAT-RIG-I-GFP and pET-RIG-I-GFP prokaryotic expression vectors were constructed. Meanwhile, they were converted to E. coli BL21 (DE3), which were induced to be expressed after culture. After the purification of His-60 nickel affinity chromatography column and the identification of SDS-PAGE, the purified TAT-RIG-I-GFP and RIG-I-GFP proteins were incubated to DF-1 cells. Finally, fluorescence microscopy was used to observe whether the corresponding fluorescence was produced in DF-1 cells. The results showed that pET-TAT-RIG-I-GFP fusion with TAT showed obvious green fluorescence in DF-1 cells. However, the pET-RIG-I-GFP without TAT cannot display green fluorescence. This shows that TAT-fused protein have successfully delivered DF-1 cells and play a key role in transmembrane delivery. In conclusion, these results provide a solid material basis for further study of antiviral drugs in poultry.
Cell Membrane ; DNA Primers ; Escherichia coli ; Gene Expression ; Gene Products, tat ; Genetic Vectors ; Recombinant Fusion Proteins

In recent years, research on immunotherapy has made great progress. Currently, immunotherapy has made significant breakthrough, especially programmed death 1/programmed death-ligand 1 (PD-1/PD-L1) checkpoint inhibitors (e.g, Nivolumab, Pembrolizumab, Atezolizumab, Durvalumab and Avelumab, etc.) have brought clinical benefits to patients with various pathological types of lung cancer, including squamous cell carcinoma, adenocarcinoma and small cell lung cancer. In this paper, the application value and current status of PD-1/PD-L1 checkpoint inhibitors in lung cancer were comprehensively analyzed by reviewing and interpreting representative clinical studies. Based on the results of various large-scale clinical trials results, the indications of immunotherapy in lung cancer have been continuously broadened, and the details of immunotherapy have also been constantly optimized. However, immunotherapy still faces many challenges, such as the selection of immune combination strategies, the exploration of biomarkers, the management of adverse events, the feasibility of application of driver gene mutation population and so on. In this article, we made a systematic review about the latest progress of PD-1/PD-L1 checkpoint inhibitors in lung cancer, in order to provide cutting-edge reference for the clinical workers.
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Animals ; Antineoplastic Agents, Immunological ; therapeutic use ; B7-H1 Antigen ; antagonists & inhibitors ; genetics ; immunology ; Humans ; Immunotherapy ; Lung Neoplasms ; drug therapy ; genetics ; immunology ; Programmed Cell Death 1 Receptor ; antagonists & inhibitors ; genetics ; metabolism

To obtain specific antibodies against nsp4 protein of porcine reproductive and respiratory syndrome virus (PRRSV), nsp4 gene was amplified by RT-PCR and cloned into pET-28a(+) vector, designated pET28a-nsp4. pET28a-nsp4 was transformed into Escherichia coli Trasseta (DE3) cells and expressed after induction of IPTG. SDS-PAGE analysis showed that the recombinant protein was expressed in soluble form with the molecular weight of 26 kDa. The soluble fusion protein in the supernatant was purified using Ni+-NTA affinity chromatography. New Zealand rabbits were immunized by the purified nsp4 and anti-sera against nsp4 were obtained. The titer of polyclonal antibodies was about 106 and showed good specificity and sensitivity in the immunofluorescence assay and Western blotting analysis. The polyclonal antibodies also recognized native nsp4 form PRRSV infected Marc-145 cells, providing a useful tool in PRRSV replication mechanism study.

Rheumatoid arthritis （RA） is a refractory autoimmune disease that can cause symmetrical polyarticular disease. The key mechanism of its occurrence and development is the dysequilibrium of helper T cell 17 （Th17）/regulatory T cell （Treg） balance. Therefore， reconstructing Th17/Treg balance may be a new strategy for the treatment of RA. Traditional Chinese medicine has significant advantages in the treatment of RA such as integrity， multi-target， multi-link and multi-path. This paper summarizes the basic and clinical studies on the regulation of Th17/Treg balance in the treatment of RA by traditional Chinese medicine in the past five years， and finds that the active components/sites of traditional Chinese medicine such as flavonoids， alkaloids and terpenes have unique advantages in the regulation of Th17/Treg balance. The traditional Chinese medicine compound formula interferes with Th17/Treg balance by exerting the effects of dispelling wind， dehumidifying， removing blood stasis， unblocking collaterals， relieving pain， dispersing cold and strengthening health. The effect of external treatment of traditional Chinese medicine is obvious and can be used as a clinical adjuvant therapy for RA； related mechanisms of action include regulating the production of inflammatory factors， regulating the expression of transcription factors and interfering with the activation of signaling pathways. However， the existing research has the shortcomings of insufficient mechanism research， few clinical research， limited external treatment research of traditional Chinese medicine， and lack of combination therapy research， which need to be improved by follow- up research.

Objective:To systematically evaluate the effect of brain-computer interface (BCI) on upper-limb motor function after stroke, and compare the effects under different interfaces. Methods:Randomized controlled trials (RCTs) about BCI for upper-limb motor function after stroke were retrieved from databases of PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang Data and CBM, from inception to October, 2020. The quality of the trials was assessed and the data were extracted according to the Cochrane Handbook of Systematic Review. A meta-analysis was carried out with RevMan 5.3 and ADDIS 1.16.8. Results:Ultimately, 14 RCTs involving 504 patients were included. Meta-analysis showed that BCI could obviously improve the Fugl-Meyer Assessment-Upper Extremities (FMA-UE) score (MD = 6.81, 95%CI 1.51 to 12.11, P < 0.05), Action Research Arm Test score (MD = 7.68, 95%CI 0.49 to 14.88, P < 0.05) and modified Barthel Index score (MD = 8.91, 95%CI 5.57 to 12.25, P < 0.001) after stroke. Subgroup analysis showed that FMA-UE score could be improved by BCI for both more than four weeks (MD = 9.44, 95%CI 1.83 to 17.04, P < 0.05) and less than four weeks (MD = 5.18, 95%CI 2.84 to 7.51, P < 0.001). For the types of interface, the probabilities of the best effects from network meta-analysis ranked as electrical stimulator (P = 0.53), visual feedback (P = 0.41) and machine assistance (P = 0.06). Conclusion:BCI, especially with electrical stimulator interface, could obviously improve upper-limb motor function and activities of daily living for stroke patients.