ABSTRACT
ObjectiveTo reveal the medication rules of the clinical prescriptions containing Citri Sarcodactylis Fructus， and to provide a basis for the modern clinical application of Citri Sarcodactylis Fructus， the development of health products and the research and development of new drugs. MethodThe clinical prescriptions containing Citri Sarcodactylis Fructus were retrieved from CNKI， Wanfang Data， and VIP， and then a basic database of prescriptions was established via the traditional Chinese medicine inheritance auxiliary platform （V2.5）， IBM SPSS V20， and Excel （Microsoft Office 2016）. The frequency and association rules of the medicines in prescriptions （compatible medicines） and the corresponding syndromes were then mined. ResultThe prescriptions were screened according to the inclusion and exclusion criteria. A total of 458 clinical prescriptions containing Citri Sarcodactylis Fructus were collected， involving 388 Chinese medicines， and the total frequency of medicines reached 6 199. The core compatible medicines （frequency > 130） of Citri Sarcodactylis Fructus included Poria （frequency of 222）， Atractylodis Macrocephalae Rhizoma （217）， Paeoniae Radix Alba （196）， Bupleuri Radix （159）， and Citri Reticulatae Pericarpium （142）. The Citri Sarcodactylis Fructus-compatible medicines with frequency > 49 were selected for further analysis， which included 34 medicines with the cumulative frequency of 3 131 （50.51% of the total frequency）. These medicines mainly have the functions of tonifying Qi， invigorating Qi， tonifying blood， alleviating edema and promoting urination， promoting digestion， and activating blood and relieving pain. They are mainly warm， cold， or mild-natured， taste bitter， sweet， or acrid， and have the tropism in the spleen， liver， stomach， or lung meridians. The association rule analysis demonstrated that 14 medicine combinations were commonly used， and the core combinations were Poria-Citri Sarcodactylis Fructus， Atractylodis Macrocephalae Rhizoma-Citri Sarcodactylis Fructus， Paeoniae Radix Alba-Citri Sarcodactylis Fructus， Bupleuri Radix-Citri Sarcodactylis Fructus， Atractylodis Macrocephalae Rhizoma-Poria-Citri Sarcodactylis Fructus， and Citri Reticulatae Pericarpium-Citri Sarcodactylis Fructus. The clinical prescriptions containing Citri Sarcodactylis Fructus were mainly used to treat 52 diseases corresponding to 11 types of traditional Chinese medicine （TCM） syndromes. Three representative syndrome types， including spleen and stomach syndromes， Qi-blood-body fluid syndromes， and gynecological syndromes were selected for further association rule analysis. In the treatment of spleen and stomach syndromes， the core compatible drugs were Atractylodis Macrocephalae Rhizoma， Poria， Paeoniae Radix Alba， Bupleuri Radix， Citri Reticulatae Pericarpium， and Pinelliae Ehizoma， which， together with Citri Sarcodactylis Fructus， formed 25 commonly used medicine combinations （16 combinations composed of 2 medicines and 9 combinations composed of 3 medicines）. In the treatment of Qi-blood-body fluid syndromes， the core compatible drugs were Poria， Paeoniae Radix Alba， Atractylodis Macrocephalae Rhizoma， Astragali Radix， Hordei Fructus Germinatus， and Bupleuri Radix， which， together with Citri Sarcodactylis Fructus， formed 23 common medicine combinations （17 combinations composed of 2 medicines， 5 combinations composed of 3 medicines， and 1 combination composed of 4 medicines）. In the treatment of gynecological syndromes， the core compatible medicines were Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， Angelicae Sinensis Radix， Astragali Radix， Cyperi Rhizoma， and Poria， which constituted 25 common medicine combinations （15 combinations composed of 2 medicines and 10 combinations composed of 3 medicines）. ConclusionWe employed the traditional Chinese medicine（TCM） inheritance auxiliary platform to explore the compatibility of Citri Sarcodactylis Fructus-containing clinical prescriptions and the corresponding TCM syndromes， which intuitively showcased the medication rules of Citri Sarcodactylis Fructus. Specifically， Citri Sarcodactylis Fructus was mainly combined with the medicines for tonifying Qi， invigorating Qi， tonifying blood， alleviating edema and promoting urination， promoting digestion， and activating blood and relieving pain to treat different TCM syndromes. While soothing liver， regulating Qi， harmonizing stomach， and relieving pain， the combinations tonify and activate blood， invigorate spleen， and resolve dampness. The findings are of great significance to the rational application of Citri Sarcodactylis Fructus， the development of health food， and the research of new drugs and will bolster the development of Chinese medicine industry.
ABSTRACT
@#AIM: To investigate the application of rectusmuscle transplantation optimization in the treatment of refractory supraventricular insufficiency in one eye caused by double supraventricular paralysis with inferior rectus restriction and high original vertical strabismus. <p>METHODS: Retrospective analysis was performed on 6 cases of double superior muscle palsy admitted to our hospital from January 2015 to January 2019. After the failure of conservative treatment, rectus muscle transplantation optimization was adopted to observe the improvement of preoperative and postoperative eye position, vertical strabismus, motor function of upper and lower rotation of paralytic eye, as well as the improvement of pseudo ptosis and compensated head position. <p>RESULTS: After the restriction was lifted, rectusmuscle transplantation optimization was performed. The original <i>in situ</i> vertical strabismus degree before surgery, one month and six months after surgery were(49.83 ± 6.55)PD,(2.67±2.07)PD, and(1.83±2.40)PD. There was a significant difference in vertical strabismus before and after 1mo and 6mo of operation(<i>P</i><0.001). There was no significant difference in the comparison of vertical strabismus between 1mo and 6mo of operation(<i>P</i>=0.899). All patients had the orthotopic position in postoperative position, and the paralysis function of the paralyzed eyes improved significantly after the operation; Pseudopterygoptosis in four cases and compensatory head position in two cases before operation were significantly improved after operation. <p>CONCLUSION: Optimization of rectus muscle transplantation is suitable for unilateral superior rotation deficiency with limitation of inferior rectus, which can effectively improve the superior rotation function of affected eye with stable curative effect.
ABSTRACT
Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.
ABSTRACT
Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.
ABSTRACT
<p><b>BACKGROUND</b>Endothelial progenitor cells (EPCs) transplantation is a promising therapeutic strategy for ischemic retinopathy. The current study aimed to establish a simple, reliable and fluorescent labeling method for tracking EPCs with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) in laser-injured mouse retina.</p><p><b>METHODS</b>EPCs were isolated from human umbilical cord blood mononuclear cells, cultivated, and labeled with various concentrations of CFSE. Based on fluorescence intensity and cell morphology, a 15 minutes incubation with 5 µmol/L CFSE at 37°C was selected as the optimal labeling condition. The survival capability and the apoptosis rate of CFSE-labeled EPCs were measured by Trypan blue staining and Annexin V/PI staining assay respectively. Fluorescence microscopy was used to observe the label stability during the extended culture period. Labeled EPCs were transplanted into the vitreous cavity of pigmented mice injured by retinal laser photocoagulation. Evans Blue angiography and flat mounted retinas were examined to track the labeled cells.</p><p><b>RESULTS</b>EPCs labeled with 5 µmol/L CFSE presented an intense green fluorescence and maintained normal morphology, with no significant changes in the survival capability or apoptosis rate after being labeled for 2 days, 1 and 4 weeks. The fluorescence intensity gradually decreased in the cells at the end of 4 weeks. Evans Blue angiography of the retina displayed the retinal capillarity network clearly and fluorescence leakage was observed around photocoagulated spots in the laser-injured mouse model. One week after transplantation of labeled EPCs, the fluorescent cells were identified around the photocoagulated lesions. Four weeks after transplantation, fluorescent tube-like structures were observed in the retinal vascular networks.</p><p><b>CONCLUSION</b>EPCs could be labeled by CFSE in vitro and monitored in vivo for at least 4 weeks, and participate in the repair of injured retinal vessels.</p>