ObjectiveThis paper aims to investigate the mechanism of Jinyang Dingtong plaster in improving the peripheral pain sensitization and synovial fibrosis in rats with knee osteoarthritis （KOA） by blocking the ion channels of transient receptor potentials （TRPs）. MethodsThe active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by using ultra-high performance liquid chromatography-electrospray ionization-quadrupole ion trap tandem mass spectrometry （UPLC-MS/MS） technology. A KOA rat model was established through intra-articular injection of monoiodoacetic acid. The rats were randomly divided into blank control group， KOA group， compound Nanxing Zhitong plaster Group， and Jinyang Dingtong plaster group， with eight rats per group. Among them， the rats in the compound Nanxing Zhitong plaster group and the Jinyang Dingtong plaster group were intervened with external application treatment. After the intervention period， the cold and mechanical stimulus pain thresholds of rats in each group were detected， and the transverse diameter of the knee joint was measured. The levels of inflammatory factors in the serum such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， nerve growth factor （NGF）， and calcitonin gene-related peptide （CGRP） were determined by enzyme-linked immunosorbent assay （ELISA）. Protein expression levels of transient receptor potential ankyrin 1 （TRPA1）， transient receptor potential melastatin 8 （TRPM8）， transient receptor potential vanilloid 1 （TRPV1）， transient receptor potential vanilloid 4 （TRPV4）， transforming growth factor-β （TGF-β）， and vascular endothelial growth factor （VEGF） in synovial tissue were detected by Western blot. Histopathological changes in synovial tissue were observed by using hematoxylin and eosin （HE）， Masson， and Sirius red staining， while the expression of type Ⅰ collagen and alpha-smooth muscle actin （α-SMA） was detected by multiplex immunofluorescence. ResultsA total of 35 active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by UPLC-MS/MS， including phenolic acids， flavonoids， quinones， alkaloids， terpenes， lignans， and coumarins. Among them， the constituents such as berberine， paeoniflorin， ferulic acid， and caffeic acid exhibit clear anti-inflammatory， analgesic， and anti-fibrotic pharmacological effects. Compared to the blank control group， rats in the KOA group showed a significant decrease in cold and mechanical stimuli pain thresholds （P<0.01）. After 14 and 28 days of Jinyang Dingtong plaster intervention， the pain threshold in this group was significantly increased compared to that in KOA group （P<0.01）， showing no significant difference from that in compound Nanxing Analgesic plaster group. Additionally， Jinyang Dingtong plaster reduced the levels of IL-1β， TNF-α， NGF， and CGRP in the serum of KOA rats （P<0.01）， lowered the expression of TRPA1， TRPM8， TRPV1， TRPV4， TGF-β， and VEGF proteins in synovial tissue （P<0.01）， improved synovial pathological damage in KOA rats， and significantly decreased fluorescence intensity of type Ⅰ collagen and α-SMA （P<0.01）. ConclusionJinyang Dingtong plaster can improve the peripheral pain sensitization and synovial fibrosis in KOA rats by downregulating the expression of ion channels of TRPs and related inflammatory and fibrotic factors.

ObjectiveThis paper aims to investigate the mechanism of Jinyang Dingtong plaster in improving the peripheral pain sensitization and synovial fibrosis in rats with knee osteoarthritis （KOA） by blocking the ion channels of transient receptor potentials （TRPs）. MethodsThe active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by using ultra-high performance liquid chromatography-electrospray ionization-quadrupole ion trap tandem mass spectrometry （UPLC-MS/MS） technology. A KOA rat model was established through intra-articular injection of monoiodoacetic acid. The rats were randomly divided into blank control group， KOA group， compound Nanxing Zhitong plaster Group， and Jinyang Dingtong plaster group， with eight rats per group. Among them， the rats in the compound Nanxing Zhitong plaster group and the Jinyang Dingtong plaster group were intervened with external application treatment. After the intervention period， the cold and mechanical stimulus pain thresholds of rats in each group were detected， and the transverse diameter of the knee joint was measured. The levels of inflammatory factors in the serum such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， nerve growth factor （NGF）， and calcitonin gene-related peptide （CGRP） were determined by enzyme-linked immunosorbent assay （ELISA）. Protein expression levels of transient receptor potential ankyrin 1 （TRPA1）， transient receptor potential melastatin 8 （TRPM8）， transient receptor potential vanilloid 1 （TRPV1）， transient receptor potential vanilloid 4 （TRPV4）， transforming growth factor-β （TGF-β）， and vascular endothelial growth factor （VEGF） in synovial tissue were detected by Western blot. Histopathological changes in synovial tissue were observed by using hematoxylin and eosin （HE）， Masson， and Sirius red staining， while the expression of type Ⅰ collagen and alpha-smooth muscle actin （α-SMA） was detected by multiplex immunofluorescence. ResultsA total of 35 active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by UPLC-MS/MS， including phenolic acids， flavonoids， quinones， alkaloids， terpenes， lignans， and coumarins. Among them， the constituents such as berberine， paeoniflorin， ferulic acid， and caffeic acid exhibit clear anti-inflammatory， analgesic， and anti-fibrotic pharmacological effects. Compared to the blank control group， rats in the KOA group showed a significant decrease in cold and mechanical stimuli pain thresholds （P<0.01）. After 14 and 28 days of Jinyang Dingtong plaster intervention， the pain threshold in this group was significantly increased compared to that in KOA group （P<0.01）， showing no significant difference from that in compound Nanxing Analgesic plaster group. Additionally， Jinyang Dingtong plaster reduced the levels of IL-1β， TNF-α， NGF， and CGRP in the serum of KOA rats （P<0.01）， lowered the expression of TRPA1， TRPM8， TRPV1， TRPV4， TGF-β， and VEGF proteins in synovial tissue （P<0.01）， improved synovial pathological damage in KOA rats， and significantly decreased fluorescence intensity of type Ⅰ collagen and α-SMA （P<0.01）. ConclusionJinyang Dingtong plaster can improve the peripheral pain sensitization and synovial fibrosis in KOA rats by downregulating the expression of ion channels of TRPs and related inflammatory and fibrotic factors.

Objective: To assess the effects of subchronic intake of high-dose Dendrobium officinale on body weight, food intake, food utilization, and blood biochemical parameters in rats, so as to provide insights into assessment of edible safety of D. officinale. Methods: Eighty SPF-grade SD rats were randomly divided into the low-, medium- and high-dose groups and the control group, of 10 male and 10 female rats in each group. Rats in the low-, medium- and high-dose groups were administered with D. officinale feeds at doses of 2.0, 4.0, and 8.0 g/kg body weight, respectively, while animals in the control group were given basic diet for successive 13 weeks. The rat body weight, food intake, food utilization, and blood biochemical parameters were compared between groups. Results: Normal diet and activity was seen in all rats, and no abnormal syndromes, signs or deaths were found during the study. There were no significant differences in rat body weight, food intake, total weight gain, total food intake, alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatinine, triacylglycerol, cholesterol, total protein, albumin, albumin/globulin ratio or blood glucose among four groups (P>0.05). The food utilization 7 weeks post-administration [(8.71%±0.78%) vs. (10.54%±1.37%), P<0.05] and the total food utilization [(18.00%±0.41%) vs. (19.51%±1.21%), P<0.05] were significantly lower in male rats in the high-dose group than in the control group. Conclusion Subchronic intake of high-dose D. officinale shows no toxicity in rats, and reduced food utilization may be associated with the health function of D. officinale in male rats.

Objective By comparing the physical properties (cell area, volume and elastic modulus) of red blood cells (RBCs) between newborn infants and the elderly over 80 years old, and correlation with the physiological and biochemical parameters such as total cholesterol and glycosylated hemoglobin, the effects of different ages and biochemical parameters on RBC physical properties were analyzed. Methods The mcropipette aspiration was used to measure the surface area, volume and elastic modulus of erythrocytes in newborn infants and the elderly over 80 years old, and the data were analyzed by statistical distribution analysis, correlation analysis and regression analysis. Results The mean values of RBC volume, surface area and elastic modulus in the elderly over 80 years old were smaller than those in newborn infants, and the mean values of RBC mechanical parameters in the same age group were not significantly different. The erythrocytes geometric parameter distribution of newborn infants was more concentrated than that of the elderly, while the elastic modulus distribution of newborn infants was more dispersed than that of the elderly. The mechanical properties of RBCs in newborn infants were highly correlated with the total cholesterol and gestational week; the mechanical properties of RBCs in the elderly were highly correlated with diastolic blood pressure and glycated hemoglobin. Conclusions There are significant differences in physical properties of RBCs between newborn infants and the elderly over 80 years old, and the biochemical parameters that affect physical properties of RBCs at different ages are also different.

Objective: To observe the effect of 5-hudroxymethyl-2-furfural (5-HMF) on glycolipid metabolism and hepatic function in mice with type 2 diabetes mellitus (T2DM) and hepatic injury. Methods: A low, a medium and a high 5-HMF dose group, a model group, and a control group were designed, with ten female ICR mice in each group. The low, medium and high dose group were given 0.27, 0.80 and 2.67 mg/kgbw 5-HMF, respectively, for 12 weeks; while the model group and the control group were given volume controlled deionized water. The model group and three dose groups were fed with high-fat and high-sugar food (36%), and the intraperitoneal injection of alloxan (60 mg/kgbw) was executed in the 10th and 11th week; the control group were fed with normal food. The body weight, blood glucose, blood lipid, and liver function of mice were determined regularly. The livers were stained by periodic acid Schiff and the changes in pathology were observed. Results: Compared with the control group, the serum levels of glucose (GLU), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) were significantly higher in the model group (P<0.05). Compared with the model group, the AST level in the low and high 5-HMF dose group, and the LDH level in the low, medium and high 5-HMF dose group, were significantly lower (P<0.05). There were no significant differences in the levels of GLU, total cholesterol, LDL-C, triacylglycerol, HDL-C and ALT between the model group and the three dose groups (P>0.05). Moderate to severe vacuolar degeneration was observed in the model group, while mild vacuolar degeneration was observed in the high dose group. Medium or large amount of hepatic glycogen granules were observed in the high dose group and the model group. Conclusion Under the conditions of this experiment, 5-HMF does not show any obvious function of reducing blood glucose and lipid in the mice with T2DM and liver injury, but show some protective effects on liver function．

We prepared a polymorphic form of valnemulin hydrogen tartrate (Form I) to overcome the instability and irritating odor of valnemulin hydrochloride that affect its use in the production and application of veterinary drugs. The physicochemical properties of Form I were characterized by scanning electron microscopy, X-ray powder diffraction, infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The results showed the crystal structure and thermal properties of Form I were very different from those of a commercially available form of valnemulin hydrogen tartrate (Form II). Form I and Form II were more stable than valnemulin hydrochloride after storage under irradiation and high humidity conditions, respectively. The solubility of Form I was 2.6 times that of Form II, and Form I was selected for use in pharmaceutical kinetics experiments in vivo. Compared to valnemulin hydrochloride, after oral administration at a dose of 10 mg/kg in pigs, Form I had similar pharmaceutical kinetic behavior but a slightly higher area under the concentration–time curve from time zero to the last measurable concentration. Consequently, Form I should be suitable for the development of simple formulations and be effective in the clinical application of veterinary drugs.
Administration, Oral ; Calorimetry, Differential Scanning ; Humidity ; Hydrogen ; Kinetics ; Microscopy, Electron, Scanning ; Odors ; Pharmacokinetics ; Powder Diffraction ; Solubility ; Spectrum Analysis ; Swine ; Veterinary Drugs

The core of medical humanity is to take care of patients. Many physicians treat patients at the bedside while trying to explore innovations to overcome the problems and obstacles that lie ahead of the patient. They are tracing the international front, learning to explore, promoting the growth of knowledge, increasing the clinical skills. They implement common induction and identification in clinical work, and strive for the personalized treatment of patients. They follow translational medicine, engage in research pro-jects, so that novel theories can be applied to technical practice, and promote the development and break-through of medical humanistic care.

Objective To investigate the effect of paclitaxel drug-coated balloon (DCB) dilatation in treating coronary in-stent restenosis (ISR) occurring after drug-eluting stent (DES) implantation,and to observe the long-term changes of the target vascular lumen area in order to clarify the curative effect of paclitaxel DCB in treating ISR.Methods Four patients with ISR whose clinical condition met the DCB indication were selected.According to the standard process,sufficient pre-expansion of ISR was performed first,then paclitaxel DCB dilatation was carried out to dilate the lesion segment of ISR,and no stent was implanted.Both coronary angiography and intravascular ultrasound (IVUS) were performed immediately after the treatment as well as 9 months to measure the minimum lumen area (MLA) and cross-sectional area (SA) of the stent,and the intimal hyperplasia was also been evaluated.Results In all 4 patients,angiography performed immediately after paclitaxel DCB dilatation showed that neither dissection of the dilated segment of the target artery nor obvious residual stenosis was observed.Angiography performed 9 months after the treatment revealed that all dilated segments of the target arteries were patent,and no pronounced restenosis of stent segment was seen.IVUS examination was indicated that MLA became enlarged,SA showed an increasing trend,and intimal hyperplasia showed a tendency to be inhibited.Conclusion For the treatment of ISR,pure paclitaxel DCB dilatation can obtain long-term lumen area enlargement,thus,repeated stent implantation can be avoided,which,in turn,can reduce the risk of ISR recurrence.Paclitaxel DCB dilatation can locally release paclitaxel,which has curative effect on the coronary artery wall to inhibit the excessive proliferation of intima.

Objective To review multi-center experience on rescue use of extracorporeal membrane oxygenation(ECMO) in pediatric patients with respiratory failure and to investigate its short-term outcome in China.Methods The survey was conducted in 4 tertiary hospitals in China mainland.All children<18 years old who had been supported with non-open chest ECMO to rescue respiratory failure in PICU were reviewed.Results Twenty-one patients with ECMO support were diagnosed respiratory failure.Male was 14,female was 7.Twelve patients successfully weaned off ECMO, 8 patients survived to discharge and 13 died.The largest part of protopathy was severe pneumonia(33%)and acute respiratory distress syndrome(29%).Their mean age was 29.0(9.0,81.5)months, mean weight was 12.0(9.0,20.8)kg,and mean OI index was 31.5(19.2,41.0).The average length of ECMO run was 149.0(91.2,242.0)hours.The blood gas analysis showed that PO2 increased from 49.5(40.4,61.9)mmHg(1mmHg=0.133kPa) at the beginning to 65.0(42.6,120.5)mmHg at 24h after ECMO treatment,and the increase of PO2 was higher in the survival compared with the nonsurvival[52.0(1.8,89.4) mmHg vs.8.2(-15.1,33.9) mmHg,P=0.036].The SO2 increased from 80.0%(70.4%,91.8%)at ECMO initiation to 98.0%(95.6%,100%)at 24h after ECMO treatment,and increase of SO2 was higher in survival group compared to that of nonsurvival group[23.5%(11.4%,27.1%) vs.4.3%(2.4%,23.8%),P=0.039].VV mode had higher survival rate than VA mode(3/3 vs.5/18).The longer the use of ventilator before ECMO,the mortality rate increased as well.The mechanical ventilation time before ECMO was significantly longer in nonsurvival group than that in the survival group[4.5(2.5,12.0)h vs.1.6(1.0,2.2)h,P=0.015].The most common complications during ECMO run were bleeding and disfunction of oxygenator.Conclusion ECMO is an effective support treatment for the pediatric patients with respiratory failure,which significantly improves oxygenation.

BACKGROUNDAltered immunoresponse is associated with tumorigenesis and cancer progression. This study assessed the levels of tumor-infiltrating CD3 + or CD8 + T lymphocytes and interleukin-2 (IL-2) protein in radically resected non-small cell lung cancer (NSCLC) tissues to predict overall survival (OS) of the patients.

METHODSParaffin-embedded tissue specimens from 129 NSCLC patients were retrospectively collected for immunostaining of CD8 + , CD3 + , and IL-2 expression. Clinicopathological and survival data were collected and analyzed using the Chi-squared test, Kaplan-Meier curves, and the log-rank test or the Cox regression model.

RESULTSThe data showed a significant inverse association between CD8 + T lymphocyte levels and IL-2 expression (r = -0.927; P = 0.000) and between the levels of CD8 + and CD3 + T lymphocytes (r = -0.722; P = 0.000), but a positive association between CD3 + T lymphocyte levels and IL-2 expression (r = 0.781; P = 0.000) in NSCLC tissues. Furthermore, the levels of CD3 + and CD8 + T lymphocytes and IL-2 expression were associated with tumor stage (P = 0.023, 0.006, and 0.031, respectively) and the level of CD8 + T lymphocytes was associated with the patient gender (P = 0.024). In addition, the levels of CD8 + T lymphocytes were associated with an unfavorable 5-year OS, whereas patients with high levels of CD3 + T lymphocytes in tumor lesions and IL-2-expressing tumors had significantly better 5-year OS rates than patients with low levels.

CONCLUSIONSThe levels of CD8 + T cells in tumor lesions and IL-2 expression were both independent predictors of OS for these NSCLC patients. Thus, the detection of tumor-infiltrating CD3 + or CD8 + T lymphocytes and IL-2 expression could be useful to predict the prognosis of radically resected NSCLC patients.

CD3 Complex ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Female ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Lung Neoplasms ; immunology ; metabolism ; Lymphocytes, Tumor-Infiltrating ; metabolism ; Male ; Prognosis