Total Ginsenoside Extract from Panax ginseng Enhances Neural Stem Cell Proliferation and Neuronal Differentiation by Inactivating GSK-3β / 中国结合医学杂志

OBJECTIVE@#To study the effects of total ginsenosides (TG) extract from Panax ginseng on neural stem cell (NSC) proliferation and differentiation and their underlying mechanisms.@*METHODS@#The migration of NSCs after treatment with various concentrations of TG extract (50, 100, or 200 µ g/mL) were monitored. The proliferation of NSCs was examined by a combination of cell counting kit-8 and neurosphere assays. NSC differentiation mediated by TG extract was evaluated by Western blotting and immunofluorescence staining to monitor the expression of nestin and microtubule associated protein 2 (MAP2). The GSK-3β/β-catenin pathway in TG-treated NSCs was examined by Western blot assay. The NSCs with constitutively active GSK-3β mutant were made by adenovirus-mediated gene transfection, then the proliferation and differentiation of NSCs mediated by TG were further verified.@*RESULTS@#TG treatment significantly enhanced NSC migration (P<0.01 or P<0.05) and increased the proliferation of NSCs (P<0.01 or P<0.05). TG mediation also significantly upregulated MAP2 expression but downregulated nestin expression (P<0.01 or P<0.05). TG extract also significantly induced GSK-3β phosphorylation at Ser9, leading to GSK-3β inactivation and, consequently, the activation of the GSK-3β/β-catenin pathway (P<0.01 or P<0.05). In addition, constitutive activation of GSK-3β in NSCs by the transfection of GSK-3β S9A mutant was found to significantly suppress TG-mediated NSC proliferation and differentiation (P<0.01 or P<0.05).@*CONCLUSION@#TG promoted NSC proliferation and neuronal differentiation by inactivating GSK-3β.

Efficacy observation of auricular point sticking in combination with a healthy diet for simple obesity in children / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of combining auricular point sticking and a healthy diet to treat simple obesity in children aged 6-9 years old.Methods: A total of 190 eligible obese kids were divided into an observation group and a control group using the random number table method, with 95 cases in each group. The observation group was intervened by auricular point sticking plus guide on a healthy diet, while the control group was only provided with the guide on a healthy diet. The therapeutic efficacy was observed after intervention for three consecutive months, as well as the changes in body mass (BM), body mass index (BMI), waist circumference (WC), hip circumference (HC), and subcutaneous fat thickness. Results: After the 3-month intervention, the total effective rate was 91.6％ in the observation group, versus 74.7％ in the control group, and the between-group difference was statistically significant (P<0.01); in both groups, the BM, BMI, WC, HC, and subcutaneous fat thickness all decreased significantly (P<0.05), and were lower in the observation group than in the control group, showing statistical significance (P<0.05). Conclusion: Auricular point sticking plus a healthy diet is safe and effective in treating simple obesity in children, producing more significant efficacy than healthy diet intervention alone.

Pharmacokinetic Studies of Factor VIII in Chinese Boys with Severe Hemophilia A: A Single-Center Study / 中华医学杂志(英文版)

<p><b>Background</b>Although much attention has been paid to the pharmacokinetics (PKs) of different factor VIII (FVIII) concentrates in persons with hemophilia A (HA), limited information is available in young boys with severe HA. In this study, we aimed to assess the PK parameters of FVIII products in boys with severe HA in China.</p><p><b>Methods</b>A total of 36 boys (plasma-derived [pd]-FVIII, n = 15; recombinant [r] FVIII, n = 21) were enrolled between January 2015 and May 2016 in Beijing Children's Hospital. PK characteristics of FVIII products were studied according to a reduced 4-sampling time point design (1 h, 9 h, 24 h, and 48 h postinfusion).</p><p><b>Results</b>The mean FVIII half-life (t) was 10.99 ± 3.45 h (range 5.52-20.02 h), the mean in vivo recovery (IVR) was 2.01 ± 0.42 IU/dl per IU/kg (range 1.24-3.02 IU/dl per IU/kg) and mean clearance (CL) of FVIII is 4.34 ± 1.58 ml·kg·h (range 2.29-7.90 ml·kg·h). We also analyzed the influence of several parameters that potentially modulate FVIII PK. The age was closely associated with FVIII half-life (R = 0.32, P < 0.01). The tof FVIII increased by 0.59 h per year. Besides age, von Willebrand factor antigen (VWF:Ag) also was associated with FVIII half-life (R = 0.52, P < 0.01). Patients with blood Group O had a shorter FVIII half-life than patients with non-O blood group (9.40 ± 0.68 h vs. 12.3 ± 0.79 h, t = 2.70, P = 0.01). The FVIII IVR correlated with age (R = 0.21, P < 0.01) and VWF:Ag level (R = 0.28, P < 0.01). CL rates were faster in young patients and in those with low-VWF:Ag levels. CL rates of FVIII are higher in blood Group O versus non-blood Group O persons (5.02 ± 0.38 vs. 4.00 ± 0.32 ml·kg·h, t = 2.53, P = 0.02).</p><p><b>Conclusions</b>Chinese boys with severe HA have similar PK values to other ethnic groups and large differences in FVIII PK between individual patients. Age, blood group, and VWF:Ag levels are important determining factors for FVIII CL.</p>

Effect of poly(DL-lactide-co-glycolide) on scar formation after glaucoma filtration surgery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Glaucoma filtering surgery (GFS) is the most common procedure performed in the treatment of glaucoma. Although antiscarring agents help prevent postsurgical scarring and improve glaucoma surgical outcomes, they may be associated with an increased incidence of severe and potentially blinding complications. Poly(DL-lactide-co-glycolide) (PDLLA/GA) is a bioresorbable polymer, which can be prepared with a large range of physical, mechanical, and biological properties and has been widely used in medicine, including as an absorbable suture and a drug carrier and especially as a scaffold in tissue engineering. This study aimed to evaluate the effect of PDLLA/GA on scar formation after glaucoma filtration surgery (GFS).</p><p><b>METHODS</b>Forty-eight New Zealand white rabbits were divided into two groups randomly and GFS was performed on the right eye of each. PDLLA/GA membranes were put under the sclera flap for evaluation. GFS with no membrane inserted served as control. Clinical evaluations of intraocular pressure (IOP) and the presence of a filtration bleb were performed at intervals (3 days, 1, 2, 4, 8, 12, 20, and 24 weeks) postoperatively. At each time point, three eyes per group were excised to observe histological changes such as inflammation and scar formation and the expression of collagen type IV, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1). The expression of connective tissue growth factor (CTGF) mRNA was determined by reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>The lower IOP level and an effective bleb were maintained for a long time after GFS in the PDLLA/GA group. The histological analysis showed less inflammation and scar formation, weaker expression of collagen type IV and PCNA, more intense MMP-9 and TIMP-1, slightly elevated ratio of MMP-9 and TIMP-1, and a smaller increase in CTGF mRNA postoperatively in the PDLLA/GA group but less than the control group (P < 0.05).</p><p><b>CONCLUSION</b>PDLLA/GA membranes may be promising for preventing fibrosis after GFS.</p>

In vivo biological stability of chemically pretreated silicone gel inserts intended for use in keratoprostheses / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pretreatment with chemical agents could alter the surface chemistry of the silicone gel, which makes it suitable for epithelial migration onto its surface and thus enhances the cytobiocompatibility. This study aimed to evaluate the biological response of the corneal stroma to porous silicone gel pretreated with different chemical agents in vivo.</p><p><b>METHODS</b>The porous silicone gels were treated with a mixed acid solution containing 23.2% H2SO4 and 0.8% K2Cr2O7 for 10 or 15 minutes or with 30% H2O2 for 15 minutes. Discs (4 mm in diameter) were inserted into interlamellar stromal pockets of New Zealand white rabbits and followed up for a period of 3 months. Clinical evaluations such as corneal infiltration, edema and neovascularization were performed daily. At 3 months, the fibroplasias and collagen deposition were examined under light and scanning electron microscopy (SEM) and by immunohistochemical analysis.</p><p><b>RESULTS</b>Pretreatment of the discs obviously decreased conjunctival congestion, discharge, cornea edema, and the extent of neovascularization. More fibroblasts migrated into the pretreated discs than into the control, and collagen was deposited, indicating that the biocompatibility of the corneal replacements was enhanced by the chemical pretreatments. From immunohistochemical analysis, Type I collagen deposition in the pretreated silicone discs was greater than in the control.</p><p><b>CONCLUSIONS</b>Chemical treatment of silicone gel is effective in decreasing rabbit corneal inflammation, encouraging fibroblast in-growth, and enhancing tissue compatibility. Pretreated gels show good biological stability when used as a skirt material in Keratoprosthesis (Kpros).</p>

Application of Smart Plug lacrimal plug in the treatment of dry eye / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy of Smart Plug lacrimal plug in the treatment of dry eye.</p><p><b>METHODS</b>Twenty-nine patients with dry eyes (58 eyes) were treated with Smart Plug lacrimal plug. All the patients were follow up for 6-10 months (average 7.8 months). Schirmer I test (SIT), tear break-up time (TBUT) and corneal fluorescein staining were used for evaluation of the clinical efficacy.</p><p><b>RESULTS</b>The SIT was 3.35∓2.60 mm and 6.35∓4.15 mm, TBUT was 3.14∓1.22 s and 5.46∓1.26 s, and corneal fluorescein staining was 2.15∓0.09 and 0.05∓0.01 before and after the treatment, respectively (P<0.05). The subjective symptoms were improved in most of the patients.</p><p><b>CONCLUSION</b>Smart Plug lacrimal plug is effective in the treatment of dry eye due to aqueous tear deficiency, and can be one of the choice of dry eye treatment.</p>

Triggering of toll-like receptors 2 and 4 by Aspergillus fumigatus conidia in immortalized human corneal epithelial cells to induce inflammatory cytokines / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cornea epithelial cells play early and crucial roles in the initiation of ocular surface responses to pathogens. Participation of toll-like receptor (TLR) 2 and TLR4, which are major forms of fungi receptors, may be involved in Aspergillus fumigatus induced immune responses. The objective of the present study was to examine whether inactive Aspergillus fumigatus conidia induce NF-kappaB activation and production of proinflammatory cytokines, and whether the expression of TLR2 and TLR4 were amplified by conidia in cultured immortalized human corneal epithelial cells (THCEs). This may contribute to our knowledge of the mechanism by which the host cornea can successfully defend against invasive fungi.</p><p><b>METHODS</b>Aspergillus fumigatus conidia were used to challenge THCE cells. THCE cells were harvested after 0.5, 1, 2 or 4 hours incubation. Real-time quantitative PCR was performed to determine the expression of TLR2, TLR4, TNF-alpha and IL-8. Western blotting was performed to determine the expression of NF-kappaB. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the expression of TNF-alpha and IL-8. And the release of TNF-alpha and IL-8 in the cell supernatant were also assessed by ELISA with or without pretreatment with TLR2 and TLR4 neutralizing antibodies.</p><p><b>RESULTS</b>Aspergillus fumigatus conidia elicited the expression of TLR2, TLR4, TNF-alpha and IL-8 mRNA in THCEs. Exposure of THCE cells to Aspergillus fumigatus conidia resulted in NF-kappaB activation, which increased at 30 minutes (increased from 11.35+/-2.74 in the controls to 19.12+/-3.48, P<0.05) and thereafter increased steadily up to 4 hours after challenge (P<0.01). Concomitant with NF-kappaB activation, secretion of TNF-alpha and IL-8 in conidia-challenged cells was increased in a time-dependent manner. Incubation of THCE cells with TLR2 antibody or TLR4 antibody before conidia challenge resulted in inhibition of conidia-induced TNF-alpha and IL-8 secretion (P<0.05), TLR2 antibody and TLR4 antibody together significantly increased inhibition of the conidia-induced secretion of TNF-alpha and IL-8 from THCE cells (P<0.01).</p><p><b>CONCLUSION</b>Aspergillus fumigatus conidia stimulates THCEs inflammatory response through a pathway dependent on TLR2 and TLR4 signaling.</p>

Expression profiles and function of Toll-like receptors in human corneal epithelia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Toll-like receptors play an important role in the human immune system. This study was conducted to investigate the expression profiles and function of Toll-like receptor (TLR) 1 - 9 in human corneal epithelium.</p><p><b>METHODS</b>The expression of TLR1 - 9 mRNA in 20 human donor corneal epithelia samples abraded during photorefractive keratotomy (PRK) and cultivated telomerase-immortalized human corneal epithelial cells (THCEs) was examined by semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. Human peripheral blood mononuclear cells (PBMCs) were used as positive controls. The expression of the TLR2 and TLR4 proteins was detected by Western analysis. ELISA was used to detect IL-8 secretion from THCEs challenged with ligands for TLR3 and TLR4 with and without antibody blockade.</p><p><b>RESULTS</b>The expression of TLR1 - 9 at the mRNA level was detected in the epithelia of 20 patients and in THCE. Significant differences among individuals were observed. One patient was found to lack of the expression of TLR3, 4, 6 and 8, whereas another did not express TLR5. The expression of TLR2 and TLR4 protein was detected in human corneal epithelial cells. As THCE cells express TLR1 - 9, cells were challenged with lipopolysaccharides (LPS) and poly I:C to determine whether TLR4 and TLR3 were functional. The results showed that secretion of IL-8 by cells stimulated with LPS and Poly I:C was 7 to 10 fold greater than secretion by unchallenged cells. Blocking TLR4 with an anti-TLR4 antibody significantly inhibited the LPS-induced IL-8 production by THCE (P < 0.05).</p><p><b>CONCLUSION</b>Human corneal epithelial cells express multiple TLRs and are able to recognize LPS and poly I:C. Different expression profiles among individuals suggest that differences in the susceptibilities and sensitivities to bacterial and viral infection in human populations relate to different patterns of TLR expression.</p>

The role of NF-kappaB activation in lipopolysaccharide induced keratitis in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nuclear factor-kappa B (NF-kappaB) is elevated in regulating transcription of many cytokines and inflammatory mediators. The purpose of this study was to investigate the activation and the significance NF-kappaB in lipopolysaccharide (LPS) induced keratitis.</p><p><b>METHODS</b>LPS induced keratitis model was based on Wistar rats. At 0.5, 1, 3, 6, 12, 24 or 72 hours after LPS treatment, the rat corneas were observed with a slit lamp microscope, then the rats were sacrificed and their corneas were excised for routine histological analysis. The expression of NF-kappaB was detected with immunohistochemical staining. The change of tumour necrosis factors-alpha (TNF-alpha) mRNA expression was identified by reverse transcriptase polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Histological findings demonstrated that LPS treated corneas showed significant changes in corneal structure. Corneal edema, pronounced inflammatory cells infiltration and inordinate collagen fibres were observed. Immunohistochemical results showed that the expression of NF-kappaB and its activation obviously increased after LPS treatment compared with the normal group and control group. Positive cells could be observed at 0.5 hour and peak expression of NF-kappaB was observed between 3 hours and 12 hours after infection, but returned to or approached normal level by 72 hours. RT-PCR showed that the level of TNF-alpha mRNA began to increase 0.5 hour after LPS treatment, peaked at 6 hours and then subsided by 72 hours. NF-kappaB had a positive correlation with the expression of TNF-alpha mRNA (r = 0.964, P < 0.01), both NF-kappaB and TNF-alpha had a strong positive correlation with the degree of inflammatory response in LPS treated corneas (r = 0.929, P < 0.01; r = 0.587, P < 0.05, respectively).</p><p><b>CONCLUSIONS</b>The activation of NF-kappaB was increased in LPS treated corneas and was elevated in LPS induced keratitis by promoting overexpression of TNF-alpha mRNA. NF-kappaB may play an important role in the pathogenesis of LPS-associated keratitis in rats.</p>

Effect of chemical treatment of silicon gel on tissue compatibility / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Silicon gel is unfavourable for cell attachment and growth. This study was to study if pretreating the surface of silicon gel with chemical agents affects the proliferation of epithelial cells.</p><p><b>METHODS</b>Silicon gel was made and treated with either mixed acid solution (containing 232 g/dm(3) of H(2)SO(4) and 8 g/dm(3) of K(2)Cr(2)O(7)) or 300 cm(3)/dm(3) peroxide for 5, 10, and 15 minutes or 10, 15, and 20 minutes, respectively. The cultured corneal epithelial cells were seeded onto those silicon gels and kept for 13 days. Immunohistochemical investigations were then carried out for integrin (alpha 6 or beta 4) and actin.</p><p><b>RESULTS</b>Growth of the epithelial cells in silicon gels treated with mixed acid solution for 10 minutes and 15 minutes was much significant than that in the untreated gels. After a 12-hour culture, a small number of corneal epithelial cells were proliferated on the surface of the silicon gels that had been treated with peroxide for 15 minutes. After a 3-day culture, those cells were further proliferated and fused together. The corneal epithelial cells did not grow well in the silicon gels treated with peroxide for 10 or 20 minutes. Immunostaining revealed the expression of actin and integrin alpha 6 or beta 4 on the silicon gels that were treated with mixed acid solution for 10 minutes or peroxide for 15 minutes.</p><p><b>CONCLUSION</b>Silicon gels treated either with mixed acid solution for 10 or 15 minutes or with peroxide for 15 minutes improves cell proliferation.</p>

The clinical and mammographic features of plasma cell mastitis / 中华放射学杂志

Objective To investigate the clinical and mammographic features of plasma cell mastitis.Methods Twenty-five patients(28 lesions)with histologically confirmed plasma cell mastitis, aged from 26 to 70 years(mean age 41 years),were examined with X-ray mammography.The clinical manifestations and imaging features were retrospectively reviewed.Results No case was in lactation.The painful irregular masses,ranged from 1.3 to 8cm in size,were found in 22 patients,while 3 patients with acute episode.Recurrent episodes of breast masses were noted in 4 patients.Based on the mammographic appearances,the plasma cell mastitis were classified as the following four types:inflammation-like type (2/28),ductal ectasia type(3/28),focal infiltration type(10/28)and nodular type(13/28).The valuable radiogyaphic signs:(1)An asymmetrically increased density along the lactiferous duct with a flame-like appearance,inhomogeneous low density tubular structures and scattered stick-shape calcifications.(2) Architectural distortion and oil cysts formation in adjacent area,(3)Subareolar ductal ectasia.Conclusions The clinical and mammographic characteristics of plasma cell mastitis are critical to avoiding unnecessary surgery.Histopathological result is needed for the diagnosis in patients highly suspected of malignancy.