Effect of Strength Training of Hip Muscles on Functional Ankle Instability: Evaluated with Surface Electromyography / 中国康复理论与实践

Objective:To investigate the effect of strength training of hip muscles on functional ankle instability (FAI), and be evaluated with surface electromyography (sEMG). Methods:From January, 2019 to June, 2020, 60 FAI patients were recruited in Qingdao Municipal Hospital. They were divided into control group (n = 30) and observation group (n = 30) randomly. The control group received conventional therapy, including ankle joint mobilization training, strength training and balance training for six weeks, while the observation group received strength training of hip muscles in addition. Cumberland Ankle Instability Tool (CAIT), Star Excursion Balance Test (SEBT) and sEMG were used to assess the function before and after intervention, and integrated electromyography (iEMG) was measured with sEMG. The correlation of CAIT and SEBT to iEMG was analyzed with Pearson coefficient. Results:No one dropped out. Before intervention, there was no significant difference between CAIT, SEBT and iEMG between two groups (P > 0.05). CAIT and SEBT improved significantly in both groups (t > 3.657, P < 0.001) after six-week intervention; the iEMG increased significantly in the observation group (t > 22.038, P < 0.001), while no significance was found in the control group (t < 1.916, P > 0.05); all the indexes were better in the observation group than in the control group (t > 2.125, P < 0.05). iEMG of gluteus medius and gluteus maximum correlated to CAIT and SEBT in the observation group (r = 0.712 to 0.866, P < 0.05). Conclusion:The strength training of the hip muscles could improve the ankle function of FAI patients. iEMG of gluteus medius and gluteus maximum could be a valid measure to assess the effect of strength training on FAI.

Association between Chinese Medicine Therapy and Survival Outcomes in Postoperative Patients with NSCLC: A Multicenter, Prospective, Cohort Study / 中国结合医学杂志

OBJECTIVE@#To evaluate the association between Chinese medicine (CM) therapy and disease-free survival (DFS) outcomes in postoperative patients with non-small cell lung cancer (NSCLC).@*METHODS@#This multiple-center prospective cohort study was conducted in 13 medical centers in China. Patients with stage I, II, or IIIA NSCLC who had undergone radical resection and received conventional postoperative treatment according to the National Comprehensive Cancer Network (NCCN) guidelines were recruited. The recruited patients were divided into a CM treatment group and a control group according to their wishes. Patients in the CM treatment group received continuous CM therapy for more than 6 months or until disease progression. Patients in the control group received CM therapy for less than 1 month. Follow-up was conducted over 3 years. The primary outcome was DFS, with recurrence/metastasis rates as a secondary outcome.@*RESULTS@#Between May 2013 and August 2016, 503 patients were enrolled into the cohort; 266 were classified in the CM treatment group and 237 in the control group. Adjusting for covariates, high exposure to CM was associated with better DFS [hazard ratio (HR) = 0.417, 95% confidential interval (CI): 0.307-0.567)]. A longer duration of CM therapy (6-12 months, 12-18 months, >24 months) was associated with lower recurrence and metastasis rates (HR = 0.225, 0.119 and 0.083, respectively). In a subgroup exploratory analysis, CM therapy was also a protective factor of cancer recurrence and metastasis in both stage I-IIIA (HR=0.50, 95% CI: 0.37-0.67) and stage IIIA NSCLC postoperative patients (HR = 0.48, 95% CI: 0.33-0.71), DFS was even longer among CM treatment group patients.@*CONCLUSIONS@#Longer duration of CM therapy could be considered a protective factor of cancer recurrence and metastasis. CM treatment is associated with improving survival outcomes of postoperative NSCLC patients in China. (Registration No. ChiCTR-OOC-14005398).

Analysis on prescription rules of Chen Baogui's prescriptions for treating epigastric fullness based on data mining / 中国中药杂志

To summary and analyze the prescription rules of Professor Chen Baogui, a famous traditional Chinese medicine (TCM) doctor, for treating epigastric fullness. Professor Chen Baogui's prescriptions for treating epigastric fullness were collected and the treatment data were input into traditional Chinese medicine inheritance support system (TCMISS) to analyze the rules of the prescriptions by using data mining methods. Based on the screened 214 cases, the treatment experience of Professor Chen Baogui for treating epigastric fullness was summarized and analyzed. It was found that Professor Chen gave priority to recuperation of Qi activity. The results of four properties and five tastes showed Professor Chen's medication compatibility rules: one was simultaneous use of cold and warm drugs, and the other was simultaneous use of pungent drugs for dispersion and bitter drugs for purgation. In drug use, the basic prescriptions had the efficacy of promoting Qi circulation and regulating viscera function, additionally with the drugs with functions of eliminating digestion and inducing stagnation, activating blood circulation to dissipate blood stasis, replenishing Qi and nourishing Yin, tranquilizing mind, strengthening muscles and bones according to the TCM syndrome type. The clinical experience of Professor Chen for treating epigastric fullness was objectively summarized with the help of TCMISS, which was significant for analyzing and inheriting academic thinking and medication experience from famous TCM doctors.

Expressions of HPV16/18 E6 protein and its clinical, predictive value of cervical intraepithelial neoplasia / 上海交通大学学报（医学版）

Objective·To investigate the clinical significance and predictive value of human papillomavirus (HPV) 16/18 E6 protein in the different cervical intraepithelial neoplasia. Methods·The expression of HPV16/18 E6 in 10 normal cervical tissues, 33 cervical intraepithelial neoplasiaⅠ (CINⅠ ), 31 CINⅡ- Ⅲ, 30 cervical cancers was detected by immunohistochemistry, explored the expression difference and the relationship with the clinicopathological characteristics of cervical cancer and the prognosis of different CIN. Results·The positive expression rates of HPV16/18 E6 in normal cervical tissues, CINⅠ , CINⅡ - Ⅲ and cervical cancer group were up-regulated (χ2=19.82, P=0.000). HPV16/18 E6 increased positive expression rates in the low grade and the big size tumors of cervical cancer tissues were detected (P=0.033, P=0.011). There were positive correlations between the overexpression and the pathological grade, tumor size, poor prognosis of cervical cancers respectively (r=0.456, P=0.011; r=0.578, P=0.000; r=0.645, P=0.000).The sensitivity,specificity,and diagnostic accuracy rates of HPV16/18 E6 positive expression to the progression of CINⅠ,CINⅡ-Ⅲand cervical cancer were respectively 100.00%, 62.50%, 43.75%; 96.77%, 91.30%, 92.86%; 96.97%, 83.87%, 66.67%. Conclusion·HPV16/18 E6 overexpression plays an important role in the generation, development and the poor prognosis of cervical cancer. HPV16/18 E6 has a good predictive value for the prognosis and hierarchical management of cervical diseases.

Comprehensive treatment with Chinese medicine in patients with advanced non-small cell lung cancer: A multicenter, prospective, cohort study / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine whether additional Chinese medicine (CM) could prolong survival and improve the quality of life (QOL) in patients with advanced non-small cell lung cancer (NSCLC) compared with Western medicine (WM) alone.</p><p><b>METHODS</b>This was a multicenter, prospective cohort study. A total of 474 hospitalized patients with stage III-IV NSCLC were recruited and divided into 2 groups. Patients in the WM group received radiotherapy, chemotherapy, and optimal supportive therapy according to the National Comprehensive Cancer Network (NCCN) guidelines. In the integrative medicine (IM) group, individualized CM (Chinese patent medicines and injections) and WM were administered. The primary end point was overall survival, and the secondary end points were time to disease progression, adverse events, and QOL. Follow-up clinical examinations and chest radiography were performed every 2 months.</p><p><b>RESULTS</b>The median survival was 16.60 months in the IM group and 13.13 months in the WM group (P<0.01). The incidences of loss of appetite, nausea, and vomiting in the IM group were significantly lower than those in the WM group (P<0.05). The QOL based on Functional Assessment of Cancer Therapy-Lung in the IM group was markedly higher than that in the WM group at the fourth course (P<0.05).</p><p><b>CONCLUSIONS</b>Additional CM may prolong survival and improve the QOL patients with NSCLC. The adverse effects of radio- and chemotherapy may be attenuated as CM is used in combination with conventional treatments.</p>

Influence of nm23-H1 gene silence in K562 cell on its differentiation toward megakaryocyte / 中华血液学杂志

<p><b>OBJECTIVE</b>To construct a stable nm23-H1-knock-down cell model with K562 cell line and study its differentiation toward megakaryocyte.</p><p><b>METHODS</b>Eukaryotic expression vector pSilencer 4.1-CMV-sinm23 expressing siRNA targeting nm23-H1 was transfected into K562 cells with lipofectamine2000. Cells with stably nm23-H1 silence were screened out by G418. Real-time quantitative PCR, immunocytochemistry, western blot were used to confirm the nm23-H1-knock-down K562 model. Cell differentiation capacity was detected by NBT reduction assay. Surface antigen Gp IIb-IIIa (CD41) of knock-down cells treated with phorbol 12-myristate 13-acetate was analyzed by flow cytometry. Western blot was used to detect the ERK1/2 signal pathway after the stimulation of phorbol 12-myristate 13-acetate.</p><p><b>RESULTS</b>Endogenous nm23-H1 was silenced by pSilencer 4.1-CMV-sinm23 and the silence efficiency was up to 75% and 70% in mRNA and protein levels respectively compared with the mock cells. Under phorbol 12-myristate 13-acetate treatment, the knock-down cells displayed a significantly increased differentiation ability toward megakaryocyte compared with control. The NBT reduction values were (0.31 +/- 0.07) and (0.23 +/- 0.05) respectively. Further results revealed that nm23-H1 gene regulating the megakaryocytic differentiation was due in part to the increased ERK1/2 phosphorylation.</p><p><b>CONCLUSIONS</b>A stable nm23-H1-knock-down K562 cell model is successfully constructed. nm23-H1 involves in regulating the megakaryocytic differentiation of K562 cell line.</p>

Efficient purification of recombinant human NDPK-A in pilot-scale / 生物工程学报

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) efficiently in pilot scale, cells of rhNDPK-A producing E. coli were homogenized by high pressure under 4 degrees C, 950 Pa. The insoluble debris was removed by microfiltration and the soluble portion was concentrated by ultrafiltration. The resulted crude sample was loaded on DEAE-sepharose Fast Flow. The target fraction was collected and then load on Cibacron Blue 3GA Sepharose CL-4B. Eluted with buffer containing ATP from the AC column, rhNDPK-A was polished with ultrafiltration. The results showed that after homogenized 2 rounds, 1500g cells of E. coli brought crude sample containing 47.6g NDPK-A. Treated with microfiltration and ultrafiltration, 27.3g of NDPK-A were recovered from this bacteria homogenate. After 2-step purification with column chromatography and then polished with ultrafiltration, 17.2 g rhNDPK-A were collected with purity of 96.3%. The recovery of the whole purification process was 36.2%, and the productivity of rhNDPK-A was 1.15 g per 100 g wet cells. Comparing the recovery of each purification step, it was found that the recovery of polish is higher than that of affinity chromatography, which is higher than that of ion exchange chromatography. The limit step was the process of sample pretreatment among the 4 purification steps. Combine with the fermentation results reported before, it was deduced that the productivity of rhNDPK-A was 510 mg/L. In conclusion, an easily controlled purification condition with high yield provides material for the translation researches of NDPK; In addition, it was suggested the crucial step determine the recovery of non-secretive recombinant proteins might be the process of sample pretreatment, not be the process of column chromatography.

Silencing HSV1 gD expression in cultured cells by RNA interference / 病毒学报

To explore the anti-HSV-1 effect of silencing gD gene expression by RNA interference, five 21-nucleotide duplex small interfering RNAs(siRNAs) targeting the HSV1 gD sequence were designed and the gD-EGFP fusion gene expression vector was constructed, then co-transfected into Vero cell, and screened the effective siRNA through analyzing the intensity of the EGFP fluorescence. Finally, the anti-HSV1 effect was confirmed by plaque reduction assay, real-time PCR and daughter virus titration of HSV1 infected Vero cells transfected with siRNAs. The study demonstrated that siRNAs could effectively and specifically inhibit gD gene expression in HSV1-infected cells, but only had a little effect on HSV1 infection, so taking gD as the target of siRNA against HSV1 needs further study.

Immune Responses and Histopathological Changes in Rabbits Immunized with Inactivated SARS Coronavirus / 中国病毒学

To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

Preliminary study on a new sentinel lymphoscintigraphy agent 99mTc-Rituximab for breast patient / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To develop a new imaging agent, (99m)Tc-Rituximab, for sentinel lymphoscinti-graphy and investigate its feasibility in detecting the sentinel lymph node (SLN) for breast cancer patient.</p><p><b>METHODS</b>(99m)Tc-Rituximab was prepared by introducing (99m)Tc directly into the mclecule of Rituximab, thus a new kind of lymphoscintigraphy agent ((99m)Tc-Rituximab) was produced. Five Bal b/c mice and ten breast cancer patients underwent dynamic SLN mapping procedures using (99m)Tc-Rituximab. Axillary lymph node status in breast cancer patients were evaluated with the fusion images, which was processed by computer software. With ROI technique, SLN/background ratio and the rate of SLN uptake were calculated and the dynamic curve of SLN uptake was drawn. The other 61 breast carcinoma patients underwent routine lymphatic mapping. Sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND) were both performed in all 71 patients. Fresh SLNs and all axillary nodes were sent for pathological examination using hematoxylin and eosin (H&E) staining.</p><p><b>RESULTS</b>(99m)Tc-Rituximab, with a yield of about 95.0%, can show SLN status in both mice and patients. The fusion image showed SLNs clearly, even the depth of SLNs in the axilla. The results of dynamic SLN mapping showed visible SLN starting from 10 min, up to 16 h after injecting tracer, however, secondary lymph nodes were not detectable at all. In mice, the ratio of SLN/background was 2.1, the rate of uptake was about 3.0% to 3.5%, and the equation of the curve was produced as: y = 0.036x + 2.7875, r(2) = 0.9787. For breast cancer patients, the uptake rate was different from case to case, but the cure seemed to be consistent with logarithm. The result of SLNB and ALND showed that the sensitivity and accuracy of SLNB was both 100.0%. The average number of SLNs found in each case was 2.6. Of nineteen breast cancer patients (26.8%) with lymph node metastasis, 11 were found to only have SLN metastasis.</p><p><b>CONCLUSION</b>(99m)Tc-Rituximab, as a new tracer to mapping sentinel lymph node for breast cancer patient, is able to show the status of sentinel lymph node reliably. The fusing image is easy to obtain and be interpreted by the clinician.</p>

Analysis of the relationship between nm23-H1 gene and human chronic myeloblastic leukemia using siRNA / 生物工程学报

To investigate the relationship between nm23-H1 gene and human chronic myeloblastic leukemia we designed siRNAs which target nm23-H1 gene. According to the principles of designing siRNA, we selected three siRNAs and transfected them into K562 cells by lipofectamine2000. The expression levels of nm23-H1 mRNA were detected by reverse transcriptase polymerase chain reaction after transfection for 24 hours. The expression levels of nm23-H1 protein were assayed by immunocytochemical method after transfection for 48 hours. And after transfection for 24, 48 and 72 hours, cell proliferation was determined by MTT method. Among the three siRNAs, siNM526 can effectively inhibit the expression of nm23-H1 on mRNA and protein levels. The growth of K562 cells was suppressed after transfection of siNM526. These results suggest that low expression level of nm23-H1 in K562 cells inhibited cell proliferation, namely reduced malignant degree of them. Therefore nm23-H1 gene might be a potential target of leukemia treatment.

Physical and chemical characters of recombinant human nucleoside diphosphate kinase A / 生物工程学报

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) and determine its physical and chemical characters, recombinant NDPK-A producing E. coli was cultured in 80L fermentor under high cell density culture (HCDC) conditions. The harvested cells were treated with high pressure to break the cell up, tangential-flow microfiltration to remove the bacteria debris and ultrafiltration to concentrate the filtered solution containing target protein. The crude NDPK-A was purified by ion exchange chromatography with DEAE Sepharose Fast Flow, affinity chromatography with Cibarcron Blue 3GA Sepharose CL-4B and gel filtration with Sephadex G-100. The purity of rhNDPK-A was analyzed with SDS-PAGE and RP-HPLC. The Enzymatic activity was determined with RP-HPLC. The molecular weight (MW) was measured with matrix assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS). The N-terminal residue was sequenced with Edman method. The apparent molecular weight of rhNDPK-A in solution was determined with multiangle laser light-scattering method (MALS). It was found that the purity of rhNDPK-A was 97.3% with SDS-PAGE method and 99.2% with RP-HPLC method. The specific enzymatic activity was (900 +/- 100) u/mg. The molecular weight was 17017, which was 132 less than the calculated value according to the amino acid sequence of NDPK-A. The sequencing result of rhNDPK-A revealed that its N-terminal residue was Ala, which was the second residue on N-terminal of native NDPK-A. The calculated MW of N-terminal deleted rhNDPK-A was 17017, exactly equal to the experimental value. The result of apparent MW determination revealed that rhNDPK-A formed homohexamer in solution with a MW of 102kD. These results suggested that rhNDPK-A possessed character identical to its native counterpart of assembling into hexamer. Confirming the identity of rhNDPK-A to its native counterpart provided a good foundation for drug development and mechanism study of NDPK-A.

Influence of the reductase deficient Escherichia coli on the solubility of recombinant proteins produced in it / 生物工程学报

The cytoplasm of E. coli is a reducing environment where cysteines do not engage in disulfide bonds. Any disulfide bonds that do appear are rapidly reduced through the action of disulfide reducing enzymes such as thioredoxin and glutaredoxin. To study the influence of E. coli cytoplasm on the solubility of recombinant proteins produced in it, bovine fibroblast growth factor (BbFGF), with single disulfide bond, and anti-HBsAg single-chain Fv (HBscFv), with two disulfide bonds, were selected as the pattern molecules of simple protein and complex protein, respectively. pJN98-BbFGF, a BbFGF expressing plasmid based on the vector pET3c, was constructed and transformed into normal host BL21(DE3) and a reductase deficient strain, E. coli Origami(DE3). At the same time, pQE-HBscFv, a HBscFv expressing plasmid was constructed and transformed into M15 [pREP4] and Origami(DE3). The recombinant BbFGF and HBscFv were produced in 2 types of bacteria and their solubilities and bioactivities were determined, respectively. It was found that the majority of BbFGF had formed inclusion body in the cytoplasm of BL21 (DE3) and all of them turned into soluble protein in Origami(DE3). It was also found the productivity of BbFGF in Origami (DE3) was 5% - 10% of the total protein and the value was 15% - 23% in BL21(DE3). BbFGFs produced in 2 recombinant bacteria were purified by cation exchange and heparin affinity chromatography. MTT assay revealed that the bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED50 of BbFGFs from different bacteria was 1.6ng/mL and 2.2ng/mL, respectively. As far as HBscFvs, both of them formed inclusion body in the cytoplasm of M15 [pQE-HBscFv] and Origami [pQE-HBscFv]. The inclusion body was solubilized in 6mol/L GuHCl, purified with a His-Trap column and then refolded by dialysis step-by-step against buffers containing downtrend concentration of GuHCl. Indirect ELISA was applied to determine the HBsAg binding activity of HBscFvs. It was found there was no obvious difference between the bioactivity of refolded HBscFvs produced from 2 recombinant bacteria. On the other hand, the supernatant of Origami [pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15 [pQE-HBscFv] displayed without any bioactivity. The soluble HBsFv in the cytoplasm of Origami [pQE-HBscFv] was purified by cation exchange and immobilized metal affinity chromatography (IMAC) and the yield was 1 - 2mg/L. Those results suggested that modification of the redox environment of E. coli cytoplasm greatly improved the solubility of recombinant disulfide-bonded proteins produced in it. In the next step, we had like to co-express of molecular chaperones or refoldase to raise the yield of soluble recombinant proteins, as well as optimizing the culture condition of the "oxidizing" E. coli.