Thermal stability of neuraminidase in influenza vaccine / 中国生物制品学杂志

@#Abstract：Objective To analyze the stabilities of neuraminidase（NA）in influenza vaccine at different temperatures and provide a reference for further complete understanding of overall shelf life of vaccines. Methods Monovalent bulks of influenza H1N1，H3N2 and B vaccines were stored at 4（low temperature），25（room temperature）and 37 ℃（changed temperature）for 0. 5，2，7，24 and 48 h separately，using that at 100 ℃（extreme temperature）for 1 h as control，and determined for NA activity by enzyme⁃linked lectin method. Results The NA activities of influenza H1N1 vaccines stored at 25 and 37 ℃ decreased significantly with the increasing of time. No significant decreases were observed in H3N2 and B vaccines even after storage at two non⁃storage temperatures for 48 h. However，all the NA activities of three vaccines decreased at 100 ℃. Conclusion Both H3N2 and B vaccines showed high stability at abnormal storage temperatures not more than 37 ℃，while H1N1 vaccine was relatively sensitive to the temperature for storage.

Effect of bone cement distribution on the clinical outcome of unilateral transpedicular puncture for spinal osteoporotic fractures / 中国骨伤

OBJECTIVE@#To investigate the effects of different bone cement morphology distribution on the clinical efficacy of unilateral percutaneous vertebroplasty(PVP) for spinal osteoporotic fractures.@*METHODS@#The clinical data of 66 patients with osteoporotic vertebral compression fractures received unilateral PVP treatment from January 2019 to April 2020 were retrospectively analyzed. There were 16 males and 50 females, including 83 vertebral bodies, 45 thoracic vertebrae and 38 lumbar vertebrae, and 55 patients with single-segment, 6 double-segment, 4 three-segment and 1 four-segment. The age ranged from 60 to 93 years with an average of (76.83±8.65) years. The included patients were admitted to hospital 1 to 10 days after onset, and were diagnosed by anteroposterior and lateral X-rays, MRI and bone density examination before surgery. According to the shape of bone cement in postoperative X-ray, the patients were divided into O-shaped group (28 cases) and H-shaped group (38 cases). In O-shaped group, the bone cement presented agglomeration mass distribution in the affected vertebra in postoperative X-ray while the bone cement presented disseminated honeycomb distribution in the affected vertebrae in H-shaped group. Bone cement injection volume was collected in two groups. The intraoperative bone cement leakage and postoperative adjacent vertebral fractures were observed. The VAS of the two groups before operation and 1 day, 1 month, 6 months and 1 year after operation were compared;and ODI of the two groups 1 day, 6 months and 1 year after operation were compared. The kyphosis angle and anterior height of the affected vertebrae were measured before operation and 1 week, 1 year after operation.@*RESULTS@#All 66 patients completed 1-year follow-up, and all patients healed well at the puncture site after surgery. There were 1 case and 8 cases of bone cement leakage in O-shaped group and H-shaped group during surgery respectively (P<0.05), but no serious complications occurred. One case occurred adjacent vertebral fracture in both groups during one-year follow-up (P>0.05). There was no statistical significance in injection amount of bone cement between the two groups (P>0.05). The VAS scores of O-shaped group and H-shaped group were 7.89±0.79, 2.75±1.08, 0.46±0.58, 0.36±0.49 and 8.00±1.04, 2.58±1.15, 0.53±0.56, 0.42±0.50 before operation, 1 day, 6 months, 1 year after operation respectively, and there was no statistical significance(P>0.05), and the VAS scores were 0.96±0.58 and 1.18±0.83 at 1 month after operation respectively, with statistical significance(P<0.05). The ODI scores of O-shaped group and H-shaped group were 12.43±3.78, 10.00±2.46, 8.43±1.50 and 12.11±3.68, 9.53±2.35, 8.32±1.51 at 1 day, 6 months and 1 year after surgery respectively, and there was no statistical significance between the two groups(P>0.05). There were no statistical significance in kyphotic angles and anterior height before surgery and 1 week, 1 year after surgery between two groups (P>0.05).@*CONCLUSION@#No matter the distribution of bone cement is O-shape or H-shape, it can achieve good clinical effect, and the prognosis effect is equivalent. Therefore, when performing unilateral puncture PVP surgery, it is not necessary to deliberately increase the puncture angle of the puncture needle in order to achieve the full diffusion of the affected vertebrae, so as to reduce the risk of damaging important structures and bone cement leakage.

Effects of miR-133a-5p on the proliferation and adhesion of gastric cardia adenocarcinoma cells and the polarization of M1 macrophages / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究miR-133a-5p调控血清外泌体源性纤连蛋白1（fibronectin1，FN1）的表达对胃贲门腺癌（gastric cardia adenocarcinoma，GCA）细胞增殖、黏附和M1型巨噬细胞极化的影响。方法：借助GEO数据库分析GCA组织中的差异表达基因，进行功能富集分析。采用qPCR检测FN1在GCA组织、血清、血清外泌体和细胞中的表达。向GCA患者血清外泌体中转染FN1的过表达载体及其对照质粒，向HGC-27细胞中转染miR-133a-5p模拟物及其对照，将转染后的HGC-27细胞和外泌体共培养，再将此细胞与THP-1细胞共培养。采用CCK-8和细胞黏附实验分别检测各组HGC-27细胞的增殖和黏附情况，WB法、ELISA分别检测细胞中CD86、iNOS的水平以及对巨噬细胞分泌IL-6、IL-1β的影响。采用双荧光素酶报告基因实验验证FN1 mRNA和miR-133a-5p之间的相互作用关系。结果：与健康人对照组相比，GCA组织、血清、血清外泌体和细胞中的FN1表达水平显著上调（均P<0.05），FN1高表达的血清外泌体与GCA患者的TNM分期（P=0.032 9）和淋巴结转移有关联（P=0.012 7）。富含FN1的血清外泌体能够被GCA细胞内化，与过表达FN1的外泌体共培养能够提高HGC-27细胞的增殖和黏附能力，抑制THP-1细胞中IL-6、IL-1β、CD86和iNOS的表达，抑制M1型巨噬细胞极化（（P<0.05或P<0.01）。miR-133a-5p在GCA组织和细胞中较对照组显著降低，可负调控FN1的表达，过表达miR-133a-5p能够通过降低GCA细胞增殖和黏附能力，促进IL-6、IL-1β、CD86和iNOS的表达，部分逆转FN1对GCA细胞恶性行为的促进作用（P<0.05或P<0.01）。结论：miR-133a-5p可通过抑制血清外泌体分泌FN1对GCA细胞的恶性行为起抑制作用，对M1型巨噬细胞极化起促进作用。

Qualitative and quantitative analysis of Paris polyphylla var. chinensis by UPLC-Q-TOF-MS/MS and HPLC / 中国中药杂志

Paridis Rhizoma(PR) is prepared from the dried rhizome of Paris polyphylla var. yunnanensis(PPY) or P. polyphylla var. chinensis(PPC) in Liliaceae family. The rapid development of PPY or PPC planting industry resulted from resource shortage has caused the waste of a large number of non-medicinal resources. To clarify the chemical compositions in rhizomes, fibrous roots, stems, leaves, seeds and pericarps of PPC, and explore the comprehensive application value and development prospect of these parts, the qualitative and quantitative analyses on the different parts of PPC were carried out by ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) and high performance liquid chromatography(HPLC). A total of 136 compounds were identified, including 112 steroidal saponins, 6 flavonoids, 11 nitrogen-containing compounds and 7 phytosterols. Rhizomes, fibrous roots, and seeds mainly contained protopennogenyl glycosides and pennogenyl glycosides; leaves and stems mainly contained protodiosgenyl glycosides and diosgenyl glycosides; pericarps mainly contained pennogenyl glycosides, followed by diosgenyl glycosides. The total level of four saponins was the highest in fibrous roots and rhizomes, followed by those in the pericarps and arillate seeds, and the lowest in the stems and exarillate seeds. This study can provide data support for the comprehensive development and rational application of non-medicinal parts of PPC.

Pim-1 Protects Retinal Ganglion Cells by Enhancing Their Regenerative Ability Following Optic Nerve Crush

Provirus integration site Moloney murine leukemia virus (Pim-1) is a proto-oncogene reported to be associated with cell proliferation, differentiation and survival. This study was to explore the neuroprotective role of Pim-1 in a rat model subjected to optic nerve crush (ONC), and discuss its related molecules in improving the intrinsic regeneration ability of retinal ganglion cells (RGCs). Immunofluorescence staining showed that AAV2- Pim-1 infected 71% RGCs and some amacrine cells in the retina. Real-time PCR and Western blotting showed that retina infection with AAV2- Pim-1 up-regulated the Pim-1 mRNA and protein expressions compared with AAV2-GFP group. Hematoxylin-Eosin (HE) staining, γ-synuclein immunohistochemistry, Cholera toxin B (CTB) tracing and TUNEL showed that RGCs transduction with AAV2-Pim-1 prior to ONC promoted the survival of damaged RGCs and decreased cell apoptosis. RITC anterograde labeling showed that Pim-1 overexpression increased axon regeneration and promoted the recovery of visual function by pupillary light reflex and flash visual evoked potential. Western blotting showed that Pim- 1 overexpression up-regulated the expression of Stat3, p-Stat3, Akt1, p-Akt1, Akt2 and p-Akt2, as well as βIII-tubulin, GAP-43 and 4E-BP1, and downregulated the expression of SOCS1 and SOCS3, Cleaved caspase 3, Bad and Bax. These results demonstrate that Pim-1 exerted a neuroprotective effect by promoting nerve regeneration and functional recovery of RGCs. In addition, it enhanced the intrinsic regeneration capacity of RGCs after ONC by activating Stat3, Akt1 and Akt2 pathways, and inhibiting the mitochondrial apoptosis pathways. These findings suggest that Pim-1 may prove to be a potential therapeutic target for the clinical treatment of optic nerve injury.

Characterization of the ligand binding of PGRP-L in half-smooth tongue sole (Cynoglossus semilaevis) by molecular dynamics and free energy calculation

Background: Peptidoglycan (PGN) recognition proteins (PGRPs) are important pattern recognition receptors of the host innate immune system that are involved in the immune defense against bacterial pathogens. PGRPs have been characterized in several fish species. The PGN-binding ability is important for the function of PGRPs. However, the PGRP-PGN interaction mechanism in fish remains unclear. In the present study, the 3-D model of a long PGRP of half-smooth tongue sole (Cynoglossus semilaevis) (csPGRP-L), a marine teleost with great economic value, was constructed through the comparative modeling method, and the key amino acids involved in the interaction with Lys-type PGNs and Dap-type PGNs were analyzed by molecular dynamics and molecular docking methods. Results: csPGRP-L possessed a typical PGRP structure, consisting of five ß-sheets and four α-helices. Molecular docking showed that the van der Waals forces had a slightly larger contribution than Coulombic interaction in the csPGRP-L-PGN complex. Moreover, the binding energies of csPGRP-L-PGNs computed by MM-PBSA method revealed that csPGRP-L might selectively bind both types of MTP-PGNs and MPP-PGNs. In addition, the binding energy of each residue of csPGRP-L was also calculated, revealing that the residues involved in the interaction with Lys-type PGNs were different from that with Dap-type PGNs. Conclusions: The 3-D structure of csPGRP-L possessed typical PGRP structure and might selectively bind both types of MTP- and MPP-PGNs, which provided useful insights to understanding the functions of fish PGRPs.

Subthreshold micropulse yellow laser (577 nm) for chronic central serous chorioretinopathy with foveal leakage / 眼科新进展

Objective To assess the safety and efficacy of subthreshold micropulse yellow laser (577 nm) in the treatment of chronic central serous chorioretinopathy with foveal leakage.Methods This was a prospective study of 12 patients (12 eyes) with chronic central serous chorioretinopathy (CSC).All patients had been treated using multiple spots of subthreshold micropulse yellow laser at 577 nm with a duty cycle of 5％ over areas of focal and diffuse leakage.And lweeks,1 months,3 months,and 6 months after treatment,the best corrected visual acuity (BCVA),central macular thickness (CMT) and reduction in subretinal fluid (SRF) were recorded.Results The mean BCVA measured at 6 months after laser treatment was 0.19 ±0.11,which was in comparison to 0.27 ± 0.08 before laser treatment,and the difference was statistically significant (P =0.016).The mean CMT was significantly reduced from (432.42 ±134.17) μm before laser treatment to (248.75 ±36.06) μm after 6 months (P =0.002).The mean SRF height was significantly decreased from (213.58 ± 132.60) μm at baseline to (17.25 ±21.90) μm (P =0.002).At the last follow-up,the SRF had disappeared completely in 6 out of 12 eyes,but there were still 6 eyes suffering from SRF.There was no evidence of retinal or choroidal damage during 6-month follow up.Conclusion Subthreshold micropulse yellow laser (577 nm) is an effective treatment option for chronic CSC with foveal leakage.

Risk Factors for Prognosis of Hepatocellular Carcinoma After Curative Resection In Patients with Low Hepatitis B Viral Load

ABSTRACT Background. A retrospective cohort study was conducted to investigate the effect of hepatitis B surface antigen (HBsAg) level on prognosis in low viral load (< 2000 lU/mL) patients with hepatitis B-related hepatocellular carcinoma (HCC) after curative resection. Material and methods. A total of 192 patients with low viral load who had received curative resection of pathologically confirmed HCC were analyzed to determine the factors affecting prognosis. The risk factors for survival, early and late recurrence (2 years as a cut-off) were studied. Results. The median follow-up time was 38.5 months. The overall survival rates at 1-, 3-, and 5-year after curative resection were 94.2%, 64.0%, and 45.2%, respectively. The cumulative recurrence rates at 1-, 3-, and 5-year after curative resection were 22.4%, 46.5%, and 67.0%, respectively. Patients with high serum HBsAg levels (> 250 lU/mL) had significantly lower survival rates than those with low HBsAg levels (HR: 1.517,95% Cl: 1.005-2.292, P = 0.047). Stratified analysis showed that patients with high HBsAg levels had a significantly higher late recurrence incidence than those with low HBsAg levels (HR: 2.155, 95% Cl: 1.094-4.248, P = 0.026), but did not have a significantly higher risk of early recurrence postoperatively (HR: 1.320,95% Cl: 0. 837-2.082, P = 0.233). Multivariate analysis revealed that HBsAg > 250 lU/mL was an independent risk factor associated with late recurrence (HR: 2.109, 95% Cl: 1.068-4.165, P = 0.032). Conclusions. HBsAg > 250 lU/mL at the time of tumor resection was an independent risk factor for late recurrence in low viral load HCC patients.

Clinical manifestation and laboratory examinations of invasive Scedosporium infection / 临床检验杂志

Objective To investigate the clinical manifestation and the characteristics of laboratory examinations of invasive Scedocporium infection.Methods The clinical data of 8 patients infected with Scedosporium from January 2011 to April 2017 were collected and retrospective analysis combined with related literatures was performed.Results Among the 8 patients,6 strains of S.apiospermum,1 strains of Peudallescheria boydii and 1 strains of S.prolificans were detectable.The predisposing factors of Scedosporium infection were trauma,environmental exposure and hypoimmunity.The septahypha in specimens could be direcdy observed under microscopic examination with positive rate 100％.The growth speed of cultured colony was relatively fast and the invasiveness was strong.The colony of Scedosporium displayed various forms from white cashmere to black yeast sample.The color was gradually become dark from the center of colony with lengthening time of cultivation.Scedosporium could be identified by microscopic morphology combining culture technique.Conclusion The course of invasive Scedosporium infection may progress rapidly with serious and dangerous illness state.The most common infection of Scedosporium should be induced by S.apiospernum.The knowledge and understanding for Scedosporium infection should be strengthened to improve the level of diagnosis and treatment.

Determination of Related Substances in Pharmaceutical Excipient Benzyl Alcohol by GC / 中国药师

To establish a GC method to determine the related substances in pharmaceutical excipient benzyl alcohol. Methods:A GC method was used with an Agilent DB-wax eapillary column(0. 32 mm × 30 m,1. 8 μm)and programming temperature. The initial temperature was 50℃, and then raised to 220℃ with a rate of 5℃·min-1 and maintained for 35min. The detector was FID. The temperature of the injection port was 200℃,and the detector temperature was 310℃. The results were confirmed by GC-MS. Results:Within a certain range,the peak area and concentration of every impurity had a good linear relationship (r≥0. 999 9). The recovery was between 96. 1% and 102. 7%. The quantitative limit was between 1. 37-3. 63 ng. Toluene, benzyl chloride, benzalde-hyde and benzyl ether were found out in the samples. Conclusion:The method is accurate and convenient, and suitable for the quanti-tative determination of related substances in pharmaceutical excipient benzyl alcohol.

1483 cases of paternity test with STR loci mutation / 法医学杂志

OBJECTIVE@#To observe and analyze the mutation phenomenon of 17 STR loci of PowerPlex 18D Kit in paternity test of Yunnan population.@*METHODS@#The DNA was extracted by Chelex-100 method. The PowerPlex 18D Kit was used to test 1,483 cases and their conclusions of paternity tests were verified.@*RESULTS@#In the 1,483 cases, 1,047 were parental triplet and 436 were uniparental diad. A total of 2,530 times of meiosis was observed. One STR locus mutation was observed in 24 cases. And 11 mutation loci were found in the 17 STR loci.@*CONCLUSION@#STR loci mutation is a common phenomenon. We should collect the data of STR loci mutation, choose other good polymorphism, low mutation rate of genetic markers, to ensure that the results are accurate and reliable.

Detection of epidermal growth factor receptor gene mutation in non-small cell lung cancer by allele-specific oligonucleotide-PCR and bi-loop probe specific primer quantitative PCR / 中华病理学杂志

<p><b>OBJECTIVE</b>To compare the detection sensitivity of epidermal growth factor receptor (EGFR) mutations between allele specific oligonucleotide PCR (ASO-PCR) and bi-loop probe and specific primer quantitative PCR (BPSP-qPCR).</p><p><b>METHODS</b>A total of 96 non-small cell lung cancer specimens were selected from West China Hospital from September 2009 to December 2010. ASO-PCR was developed to detect the presence of classical EGFR mutations. A total 39 available specimens were also tested by BPSP-qPCR.</p><p><b>RESULTS</b>EGFR mutation detection rate was 30.2% (26/96) by ASO-PCR. The mutation rate was higher in female than in male patients [45.5% (20/44) vs. 17.3% (9/52), P = 0.003], non-smokers than smokers [44.1% (26/59) vs. 8.1% (3/37), P < 0.001] and adenocarcinomas than other subtypes of lung cancer [37.0% (27/73) vs. 8.7% (2/23), P = 0.01]. Among mutation negative cases by ASO-PCR, BPSP-qPCR increased the rate of detection of 19-del and L858R mutation by 10.3% (4/39) in adenocarcinomas and non-smoking subset. Overall, the mutation detection rate of BPSP-qPCR was higher than that of ASO-PCR [66.7% (26/39) vs. 41.0% (16/39), P = 0.02].</p><p><b>CONCLUSION</b>BPSP-qPCR has a better detection sensitivity than that of ASO-PCR.</p>

Detection of epidermal growth factor receptor gene mutations in non-small cell lung cancer using bi-loop probe specific primer quantitative PCR / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the sensitivity of bi-loop probe and specific primer quantitative PCR (BPSP-qPCR) in the detection of epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>BPSP-qPCR was employed to examine the presence of mutations of EFGR exon 19 through 21. Correlation of the mutations with clinicopathological characteristics and types of tumor samples were performed.</p><p><b>RESULTS</b>In the cohort of 265 specimens, 30.2% (80/265) mutations were found to be 19-del and/or L858R. Females (39.7%, 31/78), non-smokers (41.0%, 43/105) and adenocarcinoma patients (37.8%, 51/135) had a higher mutation rate (P<0.05) among 184 patients whose profiles were available. T790M combined with 19-del and/or L858R accounted for 3.3% (6/184) of the mutations. Male metastatic tumors (29.6%, 8/27), pleural fluids of females (42.9%, 9/21) and non-smokers (40.7%, 11/27) were found to have higher percentage of 19-del and/or L858R mutations, in contrast, no mutations were found in the metastatic lesions of non-adenocarcinoma patients (P>0.05).</p><p><b>CONCLUSIONS</b>BPSP-qPCR is a robust method in detection of EGFR mutations with high consistency and sensitivity. The difference of EGFR mutations in primary tumors, metastatic lesions and pleural fluids suggests that EGFR tyrosine kinase inhibitors (EGFR-TKI) treatment may have variable treatment effects depending on the tumor sites.</p>

The hypoglycemic effect and mechanism of Laminaria japonica on diabetic model induced by alloxan in rats / 国际中医中药杂志

Objective To investigate the hypoglycemic effects of Laminaria japonica (L. japonica) on diabetic model induced by alloxan in rats. Methods Sixty healthy female rats were used to establish diabetic models by injecting alloxan peritoneally, and L.japonica was applied as raw materials for potential marine drugs.The levels of fasting blood glucose (FBG) were detected by automatic blood glucose device. Enzyme linkedimmunoabsorbant assay was applied to determine the insulin level in serum. The shape and structure of isletcells were observed with histopathological staining, and the expression of superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) in islet cells were detected by immunohistochemical technique. Results After the treatment, the levels of FBG of L.japonica treated group B [(9.37±1.70) mmol/LandC (9.18±1.65 ) mmol/L, F= 32.81, q=6.35～11.72, P<0.05 ] reduced, while the serum levels of insulin in treated group A, Band C (0.0378±0.0026, 0.0378±0.0027, 0.0367±0.0035) increased(F= 11.40, q=4.28～8.47, P<0.05) significantly than those of diabetic model group (0.0456 ±0.0057) . The shape and structure of islet cells improved with the up-expressing SOD(t=4.73～4.76, P<0.05)and down-expressing iNOS (t=4.81～5.30, P<0.05) in L.japonica treated group B and C than those in diabetic model group. Conclusion L.japonica might decrease the serum level of FBG through promoting the islet cell recovery by an anti-oxide effect.

Regulating effects and mechanism of Laminaria japonica on serum lipid of hyperlipidemia rats / 中国海洋药物

Objective To investigate the regulating effects and mechanism of Laminaria japonica (L.japonica) on serum lipid of hyperlipidemia rats.Methods Forty healthy female Wistar rats were used to establish hyperlipidemia models by feeding fat-rich forage,and the powder of L.japonica was applied as a supplement in forage for test groups.The levels of serum lipid including the triglyceride(TG),total cholesterol(TC),low-density lipoprotein (LDL),high-density lipoprotein(HDL) and the activities of lipoproteinesterase(LPL) and hepatic lipase(HL) were detected by biochemical assay.Results The levels of serum TG and TC in test group decreased significantly than those in pre-treated and model group (P