Agreement of corneal high-order aberrations measured by three wavefront aberrometers in myopic adults / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To evaluate the agreement of corneal high-order aberrations from Topcon KR-1W, i.Profiler and OPD-Scan Ⅲ wavefront aberrometers in myopic adults.METHODS:A prospective clinical study. A total of 92 adult patients(92 eyes)with myopia in the department of optometry, the People's Hospital of Guangxi Zhuang Autonomous Region from June to August 2022 were enrolled. The third-order and fourth-order corneal aberrations at the pupil diameter of 4 and 6mm were measured by Topcon KR-1W, i.Profiler, and OPD-Scan Ⅲ, respectively. The difference and agreement of the three aberrometers were evaluated.RESULTS: The measurements at 6mm pupil diameter were all greater than those at 4mm pupil diameter. Although there were no statistical differences in the measurements of Z-44、Z-24 by the three aberrometers at 4 pupil diameter(P>0.05), there were statistical differences in other measurements(P<0.05). The aberration results measured by the three aberrometers were statistically different at the 6mm pupil diameter(P<0.05). The 95% limit of agreement(95%LoA)of the measurements of higher-order aberration, including the third-order aberrations at 4mm pupil diameter and the third-order and fourth-order aberrations at 6mm pupil diameter(except for the Z-24)were greater than 0.1μm. The concordance correlation coefficient(Pc)was lower than 0.90, indicating a poor consistency. The correlation coefficients of corneal higher-order aberrations were significantly different among the three aberrometers at 4 and 6mm pupil diameter(r4mm=0.215～0.805, P4mm<0.05; r6mm=0.561～0.916, P6mm<0.001).CONCLUSION:There were significant differences in the measurements of the third- and fourth-order corneal aberrations at 4 and 6mm pupil diameter among Topcon KR-1W, i.Profiler, and OPD-Scan Ⅲ, and the agreements were poor, so they are not interchangeably in clinical applications.

Effect of Ginsenoside Rg1 on Biological Activity of Cryopreserved Schwann Cells and Nerve Regeneration After Allograft in Rat Sciatic Nerve / 中国实验方剂学杂志

ObjectiveTo observe the effect of ginsenoside Rg1 （G-Rg1） on the biological activity of cryopreserved Schwann cells （SCs） of the rat sciatic nerve and explore the feasibility of G-Rg1 in reducing the cryopreservation-induced injury in SCs. MethodBilateral sciatic nerves of SD rats were randomly divided into a fresh group， a blank group， and five G-Rg1 groups of different doses （1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1）. The nerves in the blank group and the G-Rg1 groups were preserved in liquid nitrogen solutions containing 0， 1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1 G-Rg1 for four weeks. The apoptosis of SCs was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）/S100 immunofluorescence staining. The expression of cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， major histocompatibility complex （MHC）-Ⅰ， and MHC-Ⅱ was detected by Western blot. Subsequently， all nerves were cultured in the incubator at 37 ℃ with 5% CO2 for 7 days. The expression of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF） was detected by Western blot. In addition， the above cryopreserved nerves in the blank group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 groups were transplanted to the Wistar rats by allografting （blank transplantation group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 transplantation groups）， and fresh sciatic nerve allograft and isograft control group were set up. Sixteen weeks after transplantation， compound muscle action potential （CMAP） and nerve conduction velocity （NCV） were measured by electrophysiology. Nerve filament （NF）200 immunofluorescence staining， transmission electron microscopy， and toluidine blue staining were used to analyze the histology of the regenerated nerves. ResultCompared with the fresh group， the blank group and the G-Rg1 groups showed increased expression of Caspase-9， Caspase-3， and the apoptosis of SCs （P<0.05，P<0.01） and decreased expression of GDNF， NGF， MHC-Ⅰ， and MHC-Ⅱ （P<0.01）. Compared with the results in the blank group， the expression of Caspase-9 and Caspase-3 decreased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups （P<0.01）， and the apoptosis of SCs was reduced in the 1×10-7-1×10-4 mol·L-1 G-Rg1 groups（P<0.05，P<0.01） and increased in the 1×10-3 mol·L-1 group （P<0.05）， while the expression of GDNF and NGF increased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups and decreased in the 1×10-3 mol·L-1 group （P<0.05）. There was no statistical significance in the expression of MHC-Ⅰ and MHC-Ⅱ between the blank group and the G-Rg1 groups. Compared with the 1×10-7 mol·L-1 and 1×10-3 mol·L-1 G-Rg1 groups， the 1×10-6 1×10-5， 1×10-4 mol·L-1 G-Rg1 groups showed decreased expression of Caspase-3 and the apoptosis of SCs （P<0.05，P<0.01） and increased expression of GDNF and NGF （P<0.05，P<0.01）. There was no statistical significance in MHC-Ⅰ and MHC-Ⅱ expression among G-Rg1 groups. Sixteen weeks after transplantation， compared with the isograft group， the blank transplantation group and the G-Rg1 transplantation groups showed decreased CMAP， NCV， myelin sheath thickness， and number of myelinated nerve fibers （P<0.01）， and the 1×10-6 and 1×10-4 mol·L-1 G-Rg1 transplantation groups showed decreased NF200 （P<0.01）. Compared with the allograft group， the blank transplantation group and the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Compared with the blank transplantation group， the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Among all groups of G-Rg1 transplantation， each index of the 1×10-5 mol·L-1 G-Rg1 transplantation group was superior to that of the 1×10-4 and 1×10-6 mol·L-1 G-Rg1 transplantation group （P<0.05）. ConclusionG-Rg1 at a certain centration can maintain the biological activity of cryopreserved SCs of rat sciatic nerve， alleviate the cryopreservation-induced injury of rat sciatic nerve， and promote nerve regeneration after allograft.

Analysis on UPLC Fingerprint and Determination of Multi-index Components in Pipa Qingfeiyin / 中国实验方剂学杂志

Objective:To establish the ultraperformance liquid chromatography （UPLC） fingerprint of Pipa Qingfeiyin substance benchmark， and to establish a quantitative analysis method for simultaneous determination of the contents of five index components， so as to provide reference for the quality control and evaluation of this famous classical formula. Method:ACQUITY UPLC^® CSH^TM C₁₈ column （2.1 mm×100 mm， 1.7 μm） was used with mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-7 min， 5%-7%A； 7-11 min， 7%-8%A； 11-22 min， 8%-14%A； 22-30 min， 14%-15%A； 30-35 min， 15%-25%A； 35-42 min， 25%-40%A； 42-45 min， 40%-50%A； 45-50 min， 50%-60%A）， the flow rate was 0.35 mL·min^-1， the column temperature was 25 ℃， the detection wavelengths were 278 nm and 248 nm. UPLC fingerprints of 15 batches of Pipa Qingfeiyin substance benchmark were established， and the "Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine" software （2012 edition） was used for similarity analysis， and the common peaks were assigned. Cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to evaluate the fingerprint data. UPLC fingerprint method was used to simultaneously determine the contents of five components in the substance benchmark. Result:The method validation of fingerprint and determination method was good， the similarities between 15 batches of Pipa Qingfeiyin substance benchmark and their control fingerprint were ≥0.997， 23 common peaks were identified and 11 chromatographic peaks were identified. CA， PCA and OPLS-DA divided 15 batches of the substance benchmark into two groups. The linear relationship of phellodendrine hydrochloride， chlorogenic acid， berberine hydrochloride， palmatine hydrochloride and ammonium glycyrrhizinate was good in a certain range of concentration （<italic>R</italic>²>0.999）， their average recovery was 96.47%-101.16%， and the contents of these five components in the substance benchmark were 0.87-2.00， 1.53-5.95， 18.45-33.97， 3.87-6.29， 1.02-4.12 mg·g^-1， respectively. Conclusion:The established UPLC fingerprint and multi-index component content determination methods have strong specificity， good resolution and high sensitivity， it can be characterized except for the Ginseng Radix et Rhizoma flavor， which can provide reference for the quality control and evaluation of Pipa Qingfeiyin compound preparation.

Establishment of UPLC Fingerprint and Quantitative Analysis of Multi-components of Shengyutang / 中国实验方剂学杂志

Objective:To establish ultra performance liquid chromatography （UPLC） fingerprint of Shengyutang and quantitative analysis method of 11 index components in this famous classical formula. Method:UPLC-diode array detector/evaporative light scattering detector （UPLC-PDA/ELSD） was used， two chromatographic conditions were established by different detectors according to the polarity of chemical components. Conditions of fingerprint 1 were as follows：ACQUITY UPLC HSS T₃ column （2.1 mm×100 mm， 1.8 µm） with the mobile phase of acetonitrile （A）-0.6% formic acid solution （C） for gradient elution （0-4 min， 0-4%A； 4-8 min， 4%A； 8-9 min， 4%-8%A； 9-14 min， 8%-9%A； 14-21 min， 9%-15%A； 21-26 min， 15%-17%A； 26-30 min， 17%-20%A； 30-35 min， 20%-32%A； 35-40 min， 32%-40%A； 40-50 min， 40%-80%A； 50-55 min， 80%A）， the flow rate of 0.3 mL·min^-1， PDA with detection wavelengths of 280 nm and 321 nm， the column temperature at 30 ℃. Conditions of fingerprint 2 were as follows：the CORTECS C₁₈ column （3.0 mm×100 mm， 2.7 µm） with the mobile phase of acetonitrile （A）-water （D） for gradient elution （0-11 min， 19%A； 11-16 min， 19%-25%A； 16-34 min， 25%-28%A； 34-47 min， 28%-47%A； 47-60 min， 47%-80%A）， the flow rate of 0.4 mL·min^-1， ELSD with drift tube temperature of 95 ℃， the carrier gas （air） flow rate of 2.0 L·min^-1， and the column temperature at 30 ℃. UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were established， and the similarity was evaluated by similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine （2012 edition） issued by the Chinese Pharmacopoeia Commission， and the contents of eleven index components in this famous classical formula were determined. Result:The similarities of UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were >0.98 by comparing with the control fingerprint， 27 and 16 common peaks were identified in fingerprint 1， 2， respectively. It was tested and verified that the precision， repeatability， stability， linear relationship and other results of this method all met the requirements of the 2020 edition of <italic>Chinese Pharmacopoeia</italic>. The contents of chlorogenic acid， ferulic acid， calycosin glucoside， verbascoside， senkyunolide I， senkyunolide H， senkyunolide A， ginsenoside Rg₁， ginsenoside Re， ginsenoside Rb₁ and astragaloside A in 15 batches of Shengyutang were 0.063-0.193， 0.509-0.638， 0.160-0.318， 0.012-0.056， 0.394-0.519， 0.110-0.143， 0.031-0.097， 0.382-0.595， 0.292-0.505， 0.590-0.803， 0.142-0.367 mg·g^-1， respectively. Conclusion:The established detection method meets the requirements of the 2020 edition of <italic>Chinese Pharmacopoeia</italic>， which can characterize the overall characteristics of chemical components in Shengyutang， and provide experimental basis for the quality standard research of this famous classical formula.

Antitumor effects of different Ganoderma lucidum spore powder in cell- and zebrafish-based bioassays / 中西医结合学报

OBJECTIVE@#Ganoderma lucidum spore (GLS) is gaining recognition as a medicinal part of G. lucidum and has been reported to possess various pharmacological properties, such as antitumor activity. In this work, wall-broken GLS powder (BGLSP) and wall-removed GLS powder (RGLSP), two kinds of GLS powder with different manufacturing techniques, were compared in terms of contents of active constituents and in vivo and in vitro antitumor effects.@*METHODS@#The ultraviolet and visible spectrophotometry method was used to determine the contents of polysaccharides and total triterpenoids in BGLSP and RGLSP. Seventeen individual triterpenoids were further quantified using ultra-high-performance liquid chromatography and quantitative analysis of multi-components by single marker. The antitumor effects of BGLSP and RGLSP were evaluated using in vitro cell viability assay against human gastric carcinoma SGC-7901, lung carcinoma A549 and lymphoma Ramos and further validated by in vivo zebrafish xenograft models with transplanted SGC-7901, A549 and Ramos.@*RESULTS@#The results showed that the contents of polysaccharides, total triterpenoids and individual triterpenoids of RGLSP were significantly higher than those of BGLSP. Although both BGLSP and RGLSP inhibited the three tumor cell lines in vitro in a dose-dependent manner, the inhibitory effects of RGLSP were much better than those of BGLSP. In the in vivo zebrafish assay, RGLSP exhibited more potent inhibitory activities against tumors transplanted into the zebrafish compared with BGLSP, and the inhibition rates of RGLSP reached approximately 78%, 31% and 83% on SGC-7901, A549 and Ramos, respectively.@*CONCLUSION@#The results indicated that the antitumor effects of GLS were positively correlated with the contents of the polysaccharides and triterpenoids and demonstrated that the wall-removing manufacturing technique could significantly improve the levels of active constituents, and thereby enhance the antitumor activity.

Effects of Renshenjian Decoction on blood and urine metabonomics in type 2 diabetes rats based on ~1H-NMR / 中国中药杂志

Proton nuclear magnetic resonance(~1H-NMR) is used to investigate the effect of Renshenjian Decoction on serum and urine metabolism of type 2 diabetic rats with insulin resistance induced by high-sugar and high-fat diet combined with low-dose streptozotocin(STZ). After the successful establishment of the insulin resistance model of type 2 diabetes, administration for 35 days, the serum and urine of rats were taken. Once the ~1H-NMR data have been collected and processed, PCA and OPLS-DA were used to analyze them. The results show that: compared with the blank group, the contents of methionine, taurine, α-glucose and β-glucose in the serum of the model group increased significantly(P<0.001), while the contents of 3-hydroxybutyric acid, lactic acid and unsaturated fatty acids decreased significantly(P<0.01). In the model group, the contents of trimethylamine oxide, glycine, α-glucose, β-glucose, taurine and phosphocholine in urine increased significantly(P<0.05), while the contents of creatine, lactic acid, acetic acid and citric acid decreased significantly(P<0.05). Compared with the model group, the contents of 3-hydroxybutyric acid and unsaturated fatty acids in serum of rats in the treatment group increased significantly(P<0.05), while the contents of taurine, α-glucose and β-glucose decreased significantly(P<0.01). In the treatment group, the contents of lactic acid, taurine and creatine in urine increased significantly(P<0.05), while the contents of trimethylamine oxide, glycine, α-glucose, β-glucose and phosphocholine decreased significantly(P<0.01). The results show that Renshenjian Decoction can regulate metabolic disorder and promote the metabolic phenotype to return to the normal range. It displayed therapeutic effect on type 2 diabetic rats with insulin resistance and provided a certain scientific basis for the biological basic research of Renshenjian Decoction by improving insulin resistance in diabetes mellitus.

Chemical Constituents of Benzoinum and Its Anti-tumor Activities in Vitro / 中国实验方剂学杂志

Objective::To identify the active anti-tumor constituents of Benzoinum according to observation of the anti-tumor effect of chemical constituents from Benzoinum in vitro. Method::The 95%ethanol extract of Benzoinum was systematically separated by silica gel column chromatography, medium pressure liquid preparation chromatography and preparation liquid chromatography, and their structures were identified by physicochemical property and spectral data. Anti-tumor activities of the compounds of Benzoinum were screened by in vitro cells including human hepatoma cells in vitro (HepG2), human lung cancer cells (A549), human cervical cancer cells (HeLa), human breast cancer cells (MCF-7) and human prostate cancer cells (PC-3). Result::Fifteen compounds were isolated from Benzoinum and identified as myricadiol(1), 3-keto-oleanonic acid(2), (4E)-1, 5-bis(4-hydroxyphenyl)-1-methoxy-2-(methoxy-methyl)-4-pentene(3a and 3b), (E)-p-coumaryl alcohol γ-Ο-methyl ether(4), sesamin(5), 5-(3″benzoyloxypropyl)-7-methoxy-2-(3′, 4′-methylenedioxy phenyl)-benzofuran(6), dibutyl phthalate(7), methyl 4-hydroxy-3-methoxybenzoate(8), p-hydroxybenzaldehyde(9), p-hydroxyacetophenone(10), acetovanillone(11), 3-oxo-olean-11, 13(18)-dien-28, 19β-olide(12), vanillin(13), benzoic acid(14), and siaresinolic acid(15). Compounds 1 to 11 were isolated from the resin of Styrax tonkinensis for the first time. A part of these compounds had good anti-tumor activities. Among them, compound 2, 12 showed a strongest activity. Conclusion::The chemical constituents of Benzoinum have good prospects for the development and application of anti-tumor drugs.

Changes of HPLC Fingerprint and Main Flavonoids Content of Aurantii Fructus Before and After Processing / 中国实验方剂学杂志

Objective::To establish HPLC fingerprints of Aurantii Fructus and its processed products, and to quantitatively analyze the contents of four flavonoids in these products. Method::HPLC was employed with Inertsil ODS-3 C18 column (4.6 mm×250 mm, 5 μm), the mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution, the detection wavelength of 283 nm, and the flow rate of 1.0 mL·min-1. HPLC fingerprints of raw products, stir-fried bran products and processing products of Aurantii Fructus were established. Similarity evaluation and cluster analysis were used to analyze the chromatographic data. At the same time, the contents of narirutin, naringin, hesperidin and neohesperidin were determined. Result::HPLC fingerprints of Aurantii Fructus and its processed products were established, taking naringin as the reference peak, 8, 15, 11 common peaks were demarcated for raw products, stir-fried bran products, processing products, respectively, the similarities of fingerprints were >0.95.Contents of the above four flavonoids in raw products were 0.574 7%, 5.986 3%, 0.302 2%and 3.574 7%, respectively. After processing, the contents of these four components in stir-fried bran products turned into 0.948 4%, 5.103 4%, 0.549 3%and 3.533 7%, their contents in processing products turned into 0.605 3%, 4.762 3%, 0.404 7%and 3.264 9%, respectively. Conclusion::The HPLC fingerprint of Aurantii Fructus changes significantly before and after processing. The contents of four flavonoids change to a certain extent before and after processing. The order of contents of narirutin and hesperidin in samples was stir-fried bran products>processing products>raw products, while the order of contents of naringin and neohesperidin was raw products>stir-fried bran products>processing products.

Effects of Endogenous Neurotrophic Factors on Nerve Regeneration after Cryopreserved Sciatic Nerve Allograft in Rats / 中国康复理论与实践

Objective:To investigate the effect of endogenous neurotrophic factor (ENTFs) on nerve regeneration after cryopreserved sciatic nerve allograft in rats. Methods:The 15-mm sciatic nerves from female Sprague-Dawley rats were placed in DMEM solution and pretreated in vitro for 1 day, 3 days, 7 days, 14 days, and 21 days at 37 ℃ with 5% CO2 (groups A, B, C, D and E) respectively. Fresh nerve group (group F) was set up. The protein expression of glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), Bcl-2, Bax, Caspase-3, major histocompatibility complex (MHC)-I and MHC-II was detected by Western blotting. The above six groups were cryopreserved in liquid nitrogen for four weeks. The living cells and dead cells of the nerves after cryopreservation were observed by Calcein-AM/propidium iodide staining. In addition, the above six cryopreserved groups and another fresh nerve group (group G) were transplanted to the Wistar rats by allografting (groups A', B', C', D', E', F' and G'). Isograft group (group H') was set up. One week after transplantation, the expression of CD8+ T cells and macrophages of the graft were observed by immunofluorescence staining, and the plasma levels of interleukin (IL)-2, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α were detected by ELISA. Twenty weeks after transplantation, the compound muscle action potential (CMAP) and nerve conduction velocity (NCV) were examined by electrophysiology. The wet weight ratio of gastrocnemius muscle was calculated by the operational side compared with the contralateral side. The expression of neurofilament protein (NF) 200 of the transplanted nerves was observed by immunofluorescence staining. The number of myelinated nerve fibers was analyzed by toluidine blue staining. The thickness of myelinated was analyzed by electron microscopy. Results:Compared with group F, the protein expression of GDNF and NGF increased in groups C, D and E (P < 0.05); the protein expression of Bcl-2 reduced and the protein expression of Bax and Caspase-3 increased in groups B, C, D, and E (P < 0.05); the protein expression of MHC-I and MHC-II decreased in all the pretreated groups (P < 0.05). Four weeks after cryopreservation, compared with groups F and G, the number of living cells decreased in groups C, D and E. One week after transplantation, compared with groups F' and G', the expression of CD8+ T cells and macrophages decreased, and the plasma concentration of IL-2 and TNF-α decreased in groups C', D' and E' (P<0.05). Twenty weeks after transplantation, CMAP, NCV, the wet weight ratio of gastrocnemius muscle, the number of axon and thickness of myelin sheath were better in groups C', D' and E' than in groups F' and G' (P<0.05), as well as the expression of NF200. Conclusion:ENTFs can be induced by pretreating the sciatic nerve in vitro. Cryopreserved sciatic nerve with high expression of ENTFs could promote nerve regeneration and functional recovery after allograft.

Effects of Triptolide on Biological Activity of Sciatic Nerve in Cold Preservation and Nerve Regeneration after Allogeneic Transplantation in Rats / 中国康复理论与实践

@#Objective To investigate the effects of triptolide (T10) on biological activity of sciatic nerve in cold preservation and nerve regeneration after allogeneic transplantation. Methods Cell Counting Kit-8 (CCK-8) was used to test the proliferation of SCs in logarithmic phase in 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution. The sciatic nerves from Sprague-Dawley rats were pretreated in 0 mol/L, 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution at 4 ℃ or 37 ℃ for 24 hours (n = 6). The expression of nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) was detected with Western blotting. Other sciatic nerve fragments were randomly divided into fresh nerve group (group A, n = 30), DMEM preservation group (group B, n = 30), T10 preservation group (group C, n = 30), T10 pretreatment DMEM preservation group (group D, n = 30) and T10 pretreatment T10 preservation (group E, n = 30), and were stored under 4 ℃ for four weeks. Calcein-AM/PI double staining laser confocal microscope and flow cytometry were used to detect the living cells and dead cells. The expression of the major histocompatibility complex (MHC)-I, MHC-II and intercellular cell adhesion molecule-1 (ICAM-1) was detected with Western blotting. The corresponding sciatic nerves were used to repaire 10 mm defects in Wistar rats (named groups A', B', C', D' and E'), and fresh sciatic nerve from Wistar rats were also used to do it (group F'). Compound muscle action potential (CMAP) and motor nerve conduction velocity (MNCV) were tested 16 weeks after transplantation, and then the grafts were observed for the nerve regeneration. Results SCs proliferated as the controls in the T10 solution with a concentration of 1×10-9 to 1×10-7 mol/L (P > 0.05). The expression of all the neurotrophic factors was more under 37 ℃ than under 4 ℃ in all the concentrations of T10 solution, and it was the most in the concentration of 1×10-8 mol/L whenever under 37 ℃ or 4 ℃ (P < 0.05). After four weeks of cold preservation, compared with groups B, C and D, the living nerve cells were the most in group E, and the expression of MHC-I, MHC-II and ICAM-1 was the least (P < 0.05). CMAP, MNCV and the never regeneration were better in group E' than in groups A', B', C' and D' (P < 0.05). A large number of myelinated nerve fibers were observed in groups E' and F', uniformity in size, wide distribution, and with myelin sheath, compared with those in groups A', B', C' and D'. Conclusion A certain concentration of T10 can induce the sciatic nerve of rats to express neurotrophic factor in vitro, which can improve the biological activity of cold preservation nerves, reduce the immunogenicity, and promote the regeneration of recipient nerve after allogeneic transplantation. It is even better to be pretreated with T10 before cold preservation.

Amniotic cells protect and repair mouse brain cells following ischemia-reperfusion injury / 中国组织工程研究

BACKGROUND:Amniotic cells are mainly composed of amniotic epithelial cells and amniotic mesenchymal cells, which have multi-differentiation potential and can be transformed into neurons as wel as synthesize and release biological y active substances and neurotrophic factors. In preliminary studies, amniotic cells that are transplanted into the brain can significantly promote the regeneration of brain neurons. OBJECTIVE:To explore the role of amniotic cells in mouse brain cells after ischemia-reperfusion injury. METHODS:The model of cerebral ischemia-reperfusion injury was established in Babl/c mice using occlusion of bilateral common carotid arteries, and then brain cells were separated from mice. Amniotic cells were isolated from mouse placenta. Brain cells from Balb/C mice co-cultured with amniotic cells served as experimental group, and brain cells cultured with PBS as control group. RESULTS AND CONCLUSION:The viability of brain cells in the experimental group was significantly higher than that in the control group (P0.05);after 48 hours co-culture, however, the necrotic rate of brain cells was significantly lower in the experimental group than the control group (P<0.05). In cellcycle, the experiment group showed increased S phase cells;while, the control group exhibited increased G 1 phase cells and decreased S phase cells. G 2 phase cells had no changes in number in both two groups. Through the above results, amnion cells can be proved to protect and promote the regeneration of brain cells of Balb/C mice with ischemia-reperfusion injury, and inhibit cellnecrosis and apoptosis.

The effects of p38MAPK and HBxAg on cell proliferation and apoptosis in human hepatocarcinogenesis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of host-derived p38 mitogen-activated protein kinase subunit 38 (p38MAPK) and the hepatitis B virus X antigen (HbxAg) on cell proliferation and apoptosis in human hepatocellular carcinoma (HCC), and to study the mechanism underlying hepatocarcinogenesis.</p><p><b>METHODS</b>Liver tissues were biopsied from healthy individuals and patients with chronic hepatitis B (CHB), liver cirrhosis, paratumor cirrhosis, and HCC. Immunohistochemical staining was used to detect expressions of HBxAg, p38MAPK, cell cycle G2/M phase-related factors (cdc25B, p34cdc2, cyclin B1), and cell proliferation factor ki-67.The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method (known as TUNEL) was used to detect apoptosis.</p><p><b>RESULTS</b>The highest rates of HBxAg were detected in CHB (65.0%) and HCC (44.4%) liver samples, and the antigen was mainly expressed in nuclei. Increasingly higher rates of p38MAPK, cdc25B, cyclin B1, and p34cdc2 expression were detected with increases in disease severity: normal liver (40.0%, 20.0%, 20.0%, and 30.0%, respectively), chronic hepatitis B (60.0%, 65.0%, 40.0%, and 50.0%, respectively), liver cirrhosis (65.0%, 75.0%, 70.0%, and 55.0%, respectively), paratumor cirrhosis (66.7%, 75.0%, 75.0%, and 63.9%, respectively), and HCC (77.8%, 80.6%, 80.6%, and 72.2%, respectively). In addition, the intracellular location of p38MAPK expression was different under different disease conditions, showing nuclear expression in CHB and liver cirrhosis samples and cytoplasmic expression in paratumor cirrhosis and HCC samples (x2 = 1.11, P more than 0.05). The proliferation index (PI) and the apoptosis index (AI) were both increased along with disease severity (normal more than CHB more than paratumor cirrhosis more than HCC) (PI: 0.0000+/-0.000, 0.0502+/-0.011, 0.0411+/-0.009, 0.0762+/-0.017; AI: 0.0351+/-0.024, 0.0607+/-0.022, 0.0562+/-0.013, 0.0716+/-0.011), with the notable exception for liver cirrhosis (PI: 0.1810+/-0.036 and AI: 0.1200+/-0.018). PI in poorly-differentiated HCC (0.2285+/-0.062) was significantly higher than in well-differentiated HCC (0.1216+/-0.032, t = 2.082, P = 0.044). AI in well-differentiated HCC (0.152+/-0.026) was significantly higher than in poorly-differentiated HCC (0.081+/-0.022, t = 2.129, P = 0.041).</p><p><b>CONCLUSIONS</b>In the process of hepatocarcinogenesis, HBxAg may cause a series of abnormal changes in cell cycle, proliferation and apoptosis by affecting the expression of p38MAPK.</p>

The application of wireless mobile information technology in pre-hospital and in-hospital emergency treatment of severe craniocerebral injury / 中国医师进修杂志

Objective To investigate the value of the wireless mobile information technology in pre-hospital and in-hospital emergency treatment of severe craniocerebral injury.Methods The clinical data of 55 severe craniocerebral injury patients with using wireless mobile information technology in ambulance from October 2010 to March 2012 (wireless group) and 50 severe craniocerebral injury patients without using wireless mobile information technology in ambulance from April 2009 to September 2010 (traditional group) were enrolled in this study.The scores of Glasgow coma scale,time from injury to being given the exact treatment,and survival rate were compared between two groups.Results The scores of GCS between two groups showed no significant difference(t =0.551,P＞ 0.05).The time from injury to being given the exact treatment in wireless group was significantly shorter than that in traditional group [(1.646 ± 0.499) h vs.(2.085 ±0.573) h,t =4.051,P＜0.05].Comparing to traditional group,the survival rate in wireless group was significantly reduced [78.2％ (43/55) vs.58.0％ (29/50),x2 =4.057,P ＜ 0.05].Conclusions Wireless mobile information technology using in pre-hospital enable in-hospital treatment in advance,shorten the waiting time to get the exact treatment.Wireless mobile information technology is the joint between pre-hospital and in-hospital and enhance the survival rate of severe craniocerebral injury.

Decreased expression of β-nerve growth factor correlated with histological changes in a cryptorchidism rat model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nerve growth factor (NGF) is well-known for its important role in the development and maintenance of the nervous system. Along with its neurotrophic role, NGF has been detected in the testis of mouse, rat and human, suggesting an additional non-neurotrophic effect in the male reproductive system. The expression of β-NGF in the undescended testes (cryptorchidism) has not been detected at present. The aim of this study was to evaluate the expression of β-nerve growth factor mRNA and protein in experimental cryptorchidism.</p><p><b>METHODS</b>A unilateral mechanical cryptorchidism model in the Sprague-Dawley rat was established and the expression of β-NGF with histologic changes in experimental cryptorchidism were investigated using one step quantitative real-time reverse transcription-polymerase chain reaction, in situ hybridization histochemistry, immunofluorescence and hematoxylin-eosin staining.</p><p><b>RESULTS</b>The expression of β-NGF mRNA and protein were both significantly decreased in the development of unmarred testis and cryptorchidism-induced testis, and the decrease of β-NGF in cryptorchidism-induced testis was far greater than that in uninjured testis.</p><p><b>CONCLUSION</b>From this investigation, we confirmed a lower expression of β-NGF in undescended testes than in the development of testis.</p>

Effect of topical tacrolimus on xenogeneic hair follicle transplantation / 中华医学美学美容杂志

Objective To study the effect of topical tacrolimus (FK506) on the survival of the xenogeneic transplanted hair follicles from human scalp to Wistar rats. Methods In our study, Wistar rats were used as recipients and human as donor. The black hair follicles of human scalp were harvested, and then the xenogeneic grafts were transplanted to white Wistar rats on the back. 20 couples of rats were divided in 2 groups: topical tacrolimus group (group A), and blank control group (group B). After operaton, we compared the survival time of hair follicles and their histologic outcomes in order to verify the practicability of xenogeneic transplantion of hair follicles, and the topical application of tacrolimus results.Results The mean survival time of group A was longer [(49. 9 ±7. 1) days] as compared to group B [(13. 1±1. 2) days]. The longest survival time was 65 days in group A and 14 days in group B, respectively. By comparison of the results we found that topical tacrolimus prolonged the survival time of the xenotransplanted hair follicles significantly and that tropical medication could not avoid rejection. Conclusions The immune privilege function dependent on the hair follicle anagen and axillary topical tacrolimus, can prolong the survival time of the xenogeneic transplanted hair follicles in rats significantly.

Role of p38MAPK in apoptosis of human paratumor cirrhosis hepatocellular cell line QSG-7701 induced by cisplatin / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the roles of p38 MAPK in apoptosis of the normal liver cell, the paratumor cirrhosis hepatocellular cell and the hepatocellular carcinoma cell.</p><p><b>METHODS</b>Three cell lines were adopted (the normal liver cell line HL-7702, the paratumor cirrhosis hepatocellular cell line QSG-7701 and the hepatocellular carcinoma cell line QGY-7703) and treated with Diamminedichloroplatin (DDP, cisplatin) and p38MAPK inhibitor SB203580. The apoptosis and cell cycles were detected by flow cytometry and electromicroscopy. The expressions of p38MAPK, CDC25B, p34cdc2 and cyclinB1 were detected by immunocytochemical staining , confocal microscopy and western blot.</p><p><b>RESULTS</b>The apoptotic rates in all three cell lines pretreated with DDP increased obviously and the rates in normal liver cells and HCC cells increased continuously even after SB203580 treatment, whereas in paratumor cirrhosis cells the rate decreased and the cell cycle stopped at S phase.</p><p><b>CONCLUSION</b>Cisplatin induces apoptosis in the paratumor cirrhosis hepatocellular cell line QSG-7701 via activation of p38MAPK pathway and it differs in the normal liver cells from the hepatocellular carcinoma cells.</p>

Effect of qi-tonifying and stasis-eliminating therapy on expression of vascular endothelial growth factor and its receptors Flt-1, Flk-1 in the brain of intracerebral hemorrhagic rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects and mechanism of qi-tonifying and stasis-eliminating (QTSE) therapy on the expression of vascular endothelial growth factor (VEGF) and its receptors Flt-1 and Flk-1 in the brains of intracerebral hemorrhagic (model) rats.</p><p><b>METHODS</b>One hundred and eighty Sprague-Dawley rats were randomly divided into six groups: the normal group (n=5), the sham-operative (SO) group (n=35), the model group (n=35), the QTSE group (n=35), the QT group (n=35) and the SE group (n=35). All the rats except those in the normal group and SO group were established into an intracerebral hemorrhage(ICH) model by intracerebral injection of collagenase type VII and the latter three were orally administered with Buyang Huanwu Decoction (a classical recipe for QTSE) or with some of its components for qi-tonification and for stasis-elimination, respectively. To the other three groups, normal saline solutions were given instead. Behavioral tests were carried out in the animals randomly chosen from each group on days 1, 2, 4, 7, 14, 21 and 28 after modeling. The expressions of VEGF, Flk-1 and Flt-1 were determined by immunohistochemistry and the number of vascular segments with positive expression in the injured brain area of the rats was calculated.</p><p><b>RESULTS</b>From day 7 onwards, the asymmetric forelimb use rate in the QTSE group recovered more significantly than that in the other model groups. In the model group, the expressions of VEGF, Flk-1 and Flt-1 appeared on day 1 and reached a peak on day 21, then weakened gradually. In the QTSE group, as compared with the other model groups, a higher level of VEGF expression was shown from day 7 (P<0.01) and a higher level of Flt-1 expression was shown from the 7th day to the 21st day (P<0.01).</p><p><b>CONCLUSION</b>QTSE therapy can up-regulate the expressions of VEGF and its receptors (Flk-1 and Flt-1) and improve the recovery of kinetic function in the ICH rats, which may be correlated with its action in modulating vascular regeneration to promote the reconstruction of microvascular networks in the damaged areas.</p>

Apoptosis of Neural Cell and Changes of Expressions of Bax,Bcl-2 Proteins after Frontal Lobe Sharp Injury in Infant Rats / 实用儿科临床杂志

Objective To investigate the alterations and molecular mechanism of neural cell apoptosis after frontal lobe sharp injury in infant rats.Methods Brain damage was induced by frontal lobe sharp injury.dUTP nick end labeling(TUNEL) method was used to detect cell apoptosis after brain injury,and immunohistochemistry was used to detect the expression of Bax,Bcl-2 proteins in and around the area of injury,respectively.Results The apoptosis cells were found as early as 1 hour and reached a peak at 24 hours post-injury,then gradually diminished until 120 hours when only few apoptosis cells presented.The expression of Bax and Bcl-2 proteins increased significantly 1 hour after frontal lobe injury.Bax protein reached its peak at 12 hours while Bcl-2 protein reached its peak at 1 hour post-injury.The ratio of Bax to Bcl-2 increased after lesion with peak at 24 hours,and then decreased thereafter.At 120 hours,very weak expression of Bax and Bcl-2 protein detected.There were few Bax and Bcl-2 positive cells in frontal lobe cortex in blank control and sham operation group.Conclusions Cell apoptosis exists after frontal lobe sharp injury in infant rats and the change of the number of apoptotic cells is correlated with time course of post-injury.The ratio of Bax to Bcl-2 may play a vital role in cell apoptosis.

Effects of naoyian serum on VEGF protein expression in cultured rat cerebral microvascular endothelial cell with hypoxia / 中南大学学报(医学版)

OBJECTIVE@#To determine the effects of naoyian (NYA) serum on the expression of vascular endothelial growth factor (VEGF) protein in cultured rat cerebral microvascular endothelial cell (RCMEC) with hypoxia.@*METHODS@#NYA serum was separated from rat heart which had been filled stomach with NYA successively for 3 days. The rat cerebral microvascular endothelial cells were taken from the Sprageu-Dawley rat brain at postborn 7 days. The rat cerebral microvascular endothelial cells were incubated at anaerobic incubator to establish the hypoxia models. The vigo of RCMEC was determined by MTT. The level of expression of VEGF protein was measured by cell immunohistochemistry and Western blot.@*RESULTS@#The OD value of NYA serum group was higher than the control groups after hypoxia for 18 hours. VEGF protein was increased by hypoxia in cerebral microvascular endothelial cells (P < 0.05). The content of VEGF protein in NYA serum containing medium was more significantly elevated than those cultured in other control media (P < 0.01).@*CONCLUSION@#VEGF protein was induced by hypoxia in rat cerebral microvascular endothelial cells, and NYA could upregulate the expression of VEGF protein, which may be one of the protection mechanisms for cerebral microvascular endothelial cells.

A study on the health status of residents affected by flood disasters / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the immediate and long-term effects of disasters caused by floods on residents health status.</p><p><b>METHODS</b>Stratified sampling by ranks of flood disaster occurred in 1996 and 1998, flood disaster areas and control areas were carried out. A retrospective study was also carried out to study all diseases involved during 1996 - 1999.</p><p><b>RESULTS</b>The incident rates of acute infectious disease in flooding areas in 1996 and 1998 were both higher than those of non-flooding areas (863.181/100 000 and 736.591/100 000, respectively). But there was no different between the incident rate of the first years in flooding areas and that of non-flooding areas. The prevalence rates of 8 kinds of chronic diseases related to circulatory system, nervous system, digestive system, injury and poisonous diseases in flooding areas were also higher than that in the non-flooding areas. The highest incidence rates of most diseases were in the mountainous flooding areas, followed by areas collapsed by flooding, and the lowest were seen in soakedareas by floods. The incidence rates of intestinal infectious diseases and respiratory infectious diseases were lower in areas where prevention and control measures were weak.</p><p><b>CONCLUSION</b>Flood could lead to the increase of incidence rates both on acute infectious diseases and non-infectious diseases. Interventions on non-infectious diseases should also be enforced to stop the epidemics when preventing and controlling acute infectious disease.</p>