Objective To observe the expressions of nucleotide-binding oligomerization domain receptor 1(NOD1)and its mediated RIP2/NF-κB signaling pathway-related proteins and autophagy-related proteins in cerebral cortex of rats with cerebral ischemia-reperfusion injury(CIRI);To explore the possible mechanism of eye acupuncture alleviating CIRI.Methods SPF-grade male Wistar rats were randomly divided into blank control group(12 rats),sham-operation group(12 rats)and modeling group(36 rats).A CIRI model was established by improved suture method.The rats in the modeling group were randomly divided into the model group,eyeacupuncture group,outside the acupoint area group,with 12 rats in each group.Neurological deficits in rats were evaluated by Longa score,TTC staining was used to observe the cerebral infarction,HE staining was used to observe the morphology of ischemic brain tissue,electron microscopy was used to observe the formation of autophagosome in ischemic brain tissue,RT-qPCR was used to detect the mRNA expressions of NOD1,receptor interacting protein 2(RIP2),nuclear factor-κB(NF-κB)p65 in ischemic cerebral cortex,Western blot was used to detect the expressions of NOD1,RIP2,NF-κB p65 and autophagy related proteins in ischemic cerebral cortex.Results Compared with the sham-operation group,the neurological deficit score of rats in the model group significantly increased(P<0.01),the infarct volume significantly increased(P<0.01),the typical cribriform infarct foci and multiple autophagosomes appeared in the ischemic brain tissue,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly increased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly increased(P<0.01),and the protein expression of p62 significantly decreased(P<0.01).Compared with the model group,the neurological deficit score in eye acupuncture group significantly decreased(P<0.01),the cerebral infarction volume significantly decreased(P<0.01),the area of cribriform reticular infarct and the number of autophagosomes in ischemic brain tissue significantly decreased,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly decreased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly decreased(P<0.01),and the protein expression of p62 significantly increased(P<0.01).There was no statistical significance compared with outside the acupoint area group.Conclusion Eye acupuncture can attenuat the injury of neurons in CIRI rats,and its mechanism may be related to inhibiting the activation of RIP2/NF-κB signaling pathway mediated by NOD1,thereby reducing autophagy of neurons.

Objective To observe the expressions of nucleotide-binding oligomerization domain receptor 1(NOD1)and its mediated RIP2/NF-κB signaling pathway-related proteins and autophagy-related proteins in cerebral cortex of rats with cerebral ischemia-reperfusion injury(CIRI);To explore the possible mechanism of eye acupuncture alleviating CIRI.Methods SPF-grade male Wistar rats were randomly divided into blank control group(12 rats),sham-operation group(12 rats)and modeling group(36 rats).A CIRI model was established by improved suture method.The rats in the modeling group were randomly divided into the model group,eyeacupuncture group,outside the acupoint area group,with 12 rats in each group.Neurological deficits in rats were evaluated by Longa score,TTC staining was used to observe the cerebral infarction,HE staining was used to observe the morphology of ischemic brain tissue,electron microscopy was used to observe the formation of autophagosome in ischemic brain tissue,RT-qPCR was used to detect the mRNA expressions of NOD1,receptor interacting protein 2(RIP2),nuclear factor-κB(NF-κB)p65 in ischemic cerebral cortex,Western blot was used to detect the expressions of NOD1,RIP2,NF-κB p65 and autophagy related proteins in ischemic cerebral cortex.Results Compared with the sham-operation group,the neurological deficit score of rats in the model group significantly increased(P<0.01),the infarct volume significantly increased(P<0.01),the typical cribriform infarct foci and multiple autophagosomes appeared in the ischemic brain tissue,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly increased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly increased(P<0.01),and the protein expression of p62 significantly decreased(P<0.01).Compared with the model group,the neurological deficit score in eye acupuncture group significantly decreased(P<0.01),the cerebral infarction volume significantly decreased(P<0.01),the area of cribriform reticular infarct and the number of autophagosomes in ischemic brain tissue significantly decreased,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly decreased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly decreased(P<0.01),and the protein expression of p62 significantly increased(P<0.01).There was no statistical significance compared with outside the acupoint area group.Conclusion Eye acupuncture can attenuat the injury of neurons in CIRI rats,and its mechanism may be related to inhibiting the activation of RIP2/NF-κB signaling pathway mediated by NOD1,thereby reducing autophagy of neurons.

This study was designed to comprehensively characterize and identify the chemical components in traditional Chinese medicine Psoraleae Fructus by establishing an ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS) method in combination with in-house library. The chromatographic separation conditions(stationary phase, column temperature, mobile phase, and elution gradient) and key MS monitoring parameters(capillary voltage, nozzle voltage, and fragmentor) were sequentially optimized via single-factor experiments. A BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was finally adopted, with the mobile phase consisting of 0.1% formic acid in water(A) and acetonitrile(B) at the flow rate of 0.4 mL·min~(-1) and column temperature of 30 ℃. Auto MS/MS was utilized for data acquisition in both positive and negative ion modes. By comparison with reference compounds, analysis of the MS~2 fragments, in-house library retrieval and literature research, 83 compounds were identified or tentatively characterized from Psoraleae Fructus, including 58 flavonoids, 11 coumarins, 4 terpenoid phenols, and 10 others. Sixteen of them were identified by comparison with reference compounds, and ten compounds may have not been reported from Psoraleae Fructus. This study achieved a rapid qualitative analysis on the chemical components in Psoraleae Fructus, which provided useful reference for elucidating its material basis and promoting the quality control.
Chromatography, High Pressure Liquid ; Medicine, Chinese Traditional ; Tandem Mass Spectrometry ; Cell Cycle ; Coumarins

Objective:To observe the effect of Wuzi Yanzong Wan made of different processed products on the apoptosis of spermatogenic cells in rats with kidney essence deficiency， and explore its protective effect on spermatogenic cells. Method:SD rats were randomly divided into the blank group， model group， whole raw product group， pharmacopoeia group and salt-processed product group， with 8 rats in each group. The kidney essence deficiency model was replicated by giving tripterygium glycoside tablets （the dose of 20 mg·kg^-1）. The flow cytometry （FCM） was used to analysis the apoptosis of spermatogenic cells in testis， the immunohistochemistry （IHC） and Western blot were used to detect the expression levels of B-cell lymphoma-2 （Bcl-2） and Bcl-2 associated X protein （Bax） in the testis. High performance liquid chromatography （HPLC） was used to compare the contents of eight components （chlorogenic acid， ellagic acid， hyperoside， isoquercitrin， verbascoside， astragalin， kaempferol and schisandrin） in Wuzi Yanzong Wan made of different processed products， the mobile phase was composed of acetonitrile （A）-0.4% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-15%A； 5-10 min， 15%-17%A； 10-25 min， 17%A； 25-35 min， 17%-26%A； 35-60 min， 26%-56%A）， the detection wavelength was set at 254 nm. Result:Compared with the model group， the total apoptosis rate of spermatogenic cells， protein expression of Bax and Bcl-2 in each administration group were improved. Among them， the pharmacopoeia group and salt-processed product group had significant effects （P<0.01）， and the improvement effect of the pharmacopoeia group and salt-processed product group was significantly better than that of the whole raw product group （P<0.05）. The contents of chlorogenic acid， hyperoside， isoquercitrin and verbascoside in Wuzi Yanzong Wan were increased after the herbal medicines being processed with salt-water. The content of ellagic acid in the salt-processed product group increased， while it decreased in the pharmacopoeia group. The contents of verbascoside， astragalin， kaempferol and schisandrin in samples from the salt-processed product group were greater than those in samples from the pharmacopoeia group. Conclusion:Wuzi Yanzong Wan may reduce the apoptosis of spermatogenic cells in rat testis by inhibiting the expression of Bax and promoting the expression of Bcl-2， and exert its effect of nourishing kidney and enriching essence. The enhanced anti-spermatogenic effect of Wuzi Yanzong Wan after processing may be related to the changes in chemical composition content after processing.

The study aims at investigating the grade standard and the quality characteristic of Pinelliae Rhizoma for commodity specification, which provides the reference for its grade standard formulation. 42 Pinelliae Rhizoma simples were collected from 5 medicinal materials markets and 2 producing areas. Based on the previous herbalogical study and market investigation, we combined with the data analysis to select the grading indicators using SPSS software for descriptive statistical analysis, analysis of variance, K-cluster analysis and correlation analysis. According to the actual production condition, we developed the grading standards of Pinelliae Rhizoma. Moreover, we compared the internal indicators(water, total ash, leachate and guanosine) of Pinelliae Rhizoma at various grade levels, and analyzed the correlation between appearance traits and internal indicators. The herbalogical study and market research found that the Pinelliae Rhizoma was better in large, solid and white. The results from descriptive and variance analysis showed that the appearance of Pinelliae Rhizoma was significantly different in weight per grain and grain number of 500 g. Referring to the 2015 Chinese Pharmacopoeia and the production practice, we use the length, weight per grain and grain number of 500 g as the classification index of Pinelliae Rhizoma. The results from correlation analysis showed that there was no significant correlation between the appearance of Pinelliae Rhizoma and the intrinsic quality index. In addition, we found there was no significant difference in the content of the intrinsic index except for the total ash and the extract. The current study established the classification index of the product specification and grade standard of Pinelliae Rhizoma with length, weight per grain and grain number of 500 g as the index, which can provide the basis for the classification of the product specification and grade of Pinelliae Rhizoma market.
Drugs, Chinese Herbal ; standards ; Pinellia ; chemistry ; Rhizome ; chemistry

To evaluate the promoting blood circulation and removing blood stasis effects of Danshen-Honghua(DH) herb pair with different preparations (alcohol, 50% alcohol and water) on blood rheology and coagulation functions in acute blood stasis rats, and optimize the best preparation method of DH based on principal component analysis(PCA), hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods. Ice water bath and subcutaneous injection of adrenaline were both used to establish the acute blood stasis rat model. Then the blood stasis rats were administrated intragastrically with DH (alcohol, 50% alcohol and water) extracts. The whole blood viscosity(WBV), plasma viscosity(PV), erythrocyte sedimentation rate(ESR) and haematocrit(HCT) were tested to observe the effects of DH herb pair with different preparations and doses on hemorheology of blood stasis rats; the activated partial thromboplastin time(APTT), thrombin time(TT), prothrombin time(PT), and plasma fibrinogen(FIB) were tested to observe the effects of DH herb pair with different preparations on blood coagulation function and platelet aggregation of blood stasis rats. Then PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods were all used to comprehensively evaluate the total promoting blood circulation and removing blood stasis effects of DH herb pair with different preparations. The hemorheological indexes and coagulation parameters of model group had significant differences with normal blank group. As compared with the model group, the DH herb pair with different preparations at low, middle and high doses could improve the blood hemorheology indexes and coagulation parameters in acute blood stasis rats with dose-effect relation. Based on the PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods, the high dose group of 50% alcohol extract had the best effect of promoting blood circulation and removing blood stasis. Under the same dose but different preparations, 50% alcohol DH could obviously improve the hemorheology and blood coagulation function in acute blood stasis rats. These results suggested that DH herb pair with different preparations could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats, and the optimized preparation of DH herb pair on promoting blood effects was 50% alcohol extract, providing scientific basis for more effective application of the DH herb pair in modern clinic medicine.

Objective:To investigate the prevalence of volatile sulfur compounds ( VSC ) in oral cavity of preschool children, and to analyze related factors, thus to provide scientific basis for the prediction and treatment of halitosis. Methods:The VSC content ( hydrogen sulfide, methyl mercaptan, dimethyl sul-fide) of 170 preschool children (4 to 6 years old) was detected by a portable gas chromatograph Oral-ChromaTM . The status of the oral health was evaluated. The living habits and other aspects were obtained through questionnaires from the children' s parents. A soft package for social statistics version 13. 0 ( SPSS 13 . 0 ) was used in which univariate analysis and multivariate analysis were utilized to analyze the related factors of halitosis in children. Results:In the study, 34. 4% of the total subjects had excessive VSC. Hydrogen sulfide (H2S) [(1. 59 ± 2. 41) ng/10 mL] and total VSC concentration [(2. 14 ± 4. 42) ng/10 mL] in the girls were significantly higher (P<0. 05) than those in the boys. The tongue coating score had a significant positive correlation with H2S [tongue coating area (1. 68 ± 2. 48) ng/10 mL,tongue coating thickness (2. 18 ± 2. 69) ng/10 mL] and total VSC concentration [ tongue coating area, (2. 26 ± 4. 31) ng/10 mL,tongue coating thickness (2. 41 ± 3. 02) ng/10 mL , P<0. 01]. The site number of DI-S≥2 had a significant positive correlation with methyl mercaptan ( CH3 SH) and dime-thyl sulfide [(CH3)2S] concentration (P<0. 01). The concentration of H2S [(1. 19 ± 1. 62) ng/10 mL] in children, whose mother had a higher degree of education, was statistically lower (P<0. 01).The children who took dessert or sweat drinks more frequently had lower H2S [(1. 04 ± 1. 55) ng/10 mL, P<0. 05] concentration, while CH3SH and (CH3)2S concentration [(0. 29 ± 1. 92) ng/10 mL, (0. 37 ± 2. 06) ng/10 mL, P<0. 05) were higher in the children with mouth -breath habit. Conclu-sion:A high prevalence of halitosis was noted in preschool children. Gender, tongue coating index, deb-ris index-simplified, status of the mother' s education, frequency of eating dessert or sweat drinks and dry mouth breath were related with halitosis of preschool children. The different related factors resulted in changes of each VSC concentration.

BACKGROUNDIn clinical studies there is still a lot of controversy about the increased anterior and rotational stability between double-bundle (DB) and single-bundle (SB) anterior cruciate ligament (ACL) reconstruction. The aim of this study was to evaluate the clinical results of four-tunnel DB ACL reconstruction.

METHODSSixty-four consecutive patients with ACL ruptures from May 2005 to May 2006 were randomly assigned into two groups: 32 cases for SB ACL reconstruction and 32 cases for DB ACL reconstruction. Clinical data, including KT 2000, Biodex test, Lysholm score, Tegner score and IKDC score, were prospectively collected until at least 10 months post-operative.

RESULTSThe average values of KT 2000 were (1.47 +/- 1.17) mm and (1.68 +/- 1.14) mm for the SB and DB ACL reconstruction groups at 30 degrees of knee flexion (P > 0.05), and were (1.04 +/- 0.98) mm and (1.13 +/- 0.98) mm at 90 degrees of knee flexion (P > 0.05). There were also no significant differences in Lysholm score, Tegner score, IKDC score and Biodex test scores between the two groups (P > 0.05). The operation time of DB ACL reconstruction was 20 minutes longer than the SB ACL reconstruction (P < 0.05).

CONCLUSIONDouble bundle ACL reconstructions have no obvious clinical advantages over single bundle ACL reconstructions.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Female ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Prospective Studies ; Reconstructive Surgical Procedures ; methods ; Tendons ; transplantation ; Treatment Outcome ; Young Adult

BACKGROUNDDifferent kinds of minimally invasive surgery (MIS) procedures have now been used in total knee arthroplasty (TKA). Compared with traditional TKA procedure with a long skin incision, clinical studies showed MIS procedures had some advantages. Quadriceps sparing (QS) procedures are the most minimally invasive MIS procedure until now. This study was aimed to find the insertion types for Chinese patients' vastus medialis and if the QS procedure had some advantages in patients' early recovery.

METHODSBetween February 2006 and May 2007, 120 consecutive patients underwent unilateral primary TKA under general anesthesia, among whom 14 patients were lost to follow-up, the remaining 106 cases were enrolled in this study. Among the 106 cases there were 85 right knees, 21 left knees (15 men and 91 women, with a mean age of 65.1+/-7.4 years); osteoarthritis in 97 patients (91.5%) and rheumatoid arthritis in 9 patients (8.5%). MIS TKA was performed in 49 cases (MIS TKA group), while MIS-QS TKA in 57 cases (MIS-QS TKA group). During the operation, the type I, II and III insertions of the vastus medialis for all patients were recorded. Each knee was rated post-operatively according to the Hospital of Special Surgery (HSS) scoring system. Clinical follow-up was undertaken at 1 week, 2, 6, 12 and 24 weeks. Operating time and complications were recorded.

RESULTSThere was no statistically significant difference between the two groups for gender distribution, age, left or right knee incidence, pre-operative diagnosis, incidence of varus or valgus deformity. Of the 106 cases there was 1 (0.9%) case with a type I insertion of the vastus medialis, 4 (3.8%) cases with type II insertions, 101 (95.3%) cases with type III insertions. The HSS scoring was significantly different between the MIS-QS TKA group and MIS TKA group within the first two weeks post operation. From 2 weeks later to 24 weeks, no significant difference was found. The average operating time was (53.3+/-12.4) minutes in the MIS TKA group and (64.1+/-15.1) minutes in the MIS-QS TKA group (P<0.001). In the MIS-QS TKA group, 1 patient had delayed healing of the partial skin incision (1.8%). No other complications were found in either group.

CONCLUSIONSAlthough most of the Chinese patients had type III insertions of the vastus medialis, the MIS-QS TKA procedure showed less injury to the quadriceps than the standard MIS TKA and this could contribute to the earlier recovery of the patients. But a shorter skin incision and more tension on the skin may also lead to more skin complications.

Aged ; Aged, 80 and over ; Anesthesia Recovery Period ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Quadriceps Muscle ; surgery ; Time Factors

OBJECTIVETo observe the biomechanical change of the remodeling process of a four bundles model using semitendinosus tendon for the reconstruction of anterior cruciate ligament (ACL) in rabbits, and to explore the possibility of using braiding technique for improving the outcome for reconstruction of ACL.

METHODSTwo techniques were employed for the ACL reconstruction, one group with the implant of parallel bundles and the other with the braided bundles. Specimen were collected at 26 and 52 weeks after surgery respectively for biomechanical test. Gross morphology was also observed.

RESULTSThe group with the implant of the braided bundles showed a more fused rate. The biomechanical test showed that the strength of the reconstructed ACL with the unbraided bundles was 35.87% of the controlled normal ACL, while the braided bundles reached 67.20%. The strength of the graft of the 26 and 52 weeks with the braided bundles were significantly higher than that of the unbraided group, while the stiffness of the 52 weeks graft of the braided group was significantly higher than that of the unbraided group also.

CONCLUSIONSBraiding each bundle to become a single implant for reconstruction of ACL might have better outcome since it will decrease the movement between each bundles and improve the biomechanical properties.

Animals ; Anterior Cruciate Ligament ; surgery ; Biomechanical Phenomena ; Rabbits ; Tendons ; surgery ; Transplantation, Autologous