[Microfluidic Chip and Flow Cytometry for Examination of the Antiplatelet Effect of Ticagrelor].

Objective To examine the antiplatelet effect of ticagrelor by microfluidic chip and flow cytometry under shear stress in vitro. Methods Microfluidic chip was used to examine the effect of ticagrelor on platelet aggregation at the shear rates of 300/s and 1500/s.We adopted the surface coverage of platelet aggregation to calculate the half inhibition rate of ticagrelor.The inhibitory effect of ticagrelor on ADP-induced platelet aggregation was verified by optical turbidimetry.Microfluidic chip was used to construct an in vitro vascular stenosis model,with which the platelet reactivity under high shear rate was determined.Furthermore,the effect of ticagrelor on the expression of fibrinogen receptor (PAC-1) and P-selectin (CD62P) on platelet membrane activated by high shear rate was analyzed by flow cytometry. Results At the shear rates of 300/s and 1500/s,ticagrelor inhibited platelet aggregation in a concentration-dependent manner,and the inhibition at 300/s was stronger than that at 1500/s (both P<0.001).Ticagrelor at a concentration ≥4 µmol/L almost completely inhibited platelet aggregation.The inhibition of ADP-induced platelet aggregation by ticagrelor was similar to the results under flow conditions and also in a concentration-dependent manner.Ticagrelor inhibited the expression of PAC-1 and CD62P. Conclusion We employed microfluidic chip to analyze platelet aggregation and flow cytometry to detect platelet activation,which can reveal the responses of different patients to ticagrelor.

Berberine blocks inflammasome activation and alleviates diabetic cardiomyopathy via the miR­18a­3p/Gsdmd pathway.

Diabetic cardiomyopathy (DCM) is a cardiovascular disease which has been reported as a major cause of mortality worldwide for several years. Berberine (BBR) is a natural compound extracted from a Chinese herb, with a clinically reported anti­DCM effect; however, its molecular mechanisms have not yet been fully elucidated. The present study indicated that BBR markedly alleviated DCM by inhibiting IL­1ß secretion and the expression of gasdermin D (Gsdmd) at the post­transcriptional level. Considering the importance of microRNAs (miRNAs/miRs) in the regulation of the post­transcriptional process of specific genes, the ability of BBR to upregulate the expression levels of miR­18a­3p by activating its promoter (­1,000/­500) was examined. Notably, miR­18a­3p targeted Gsdmd and abated pyroptosis in high glucose­treated H9C2 cells. Moreover, miR­18a­3p overexpression inhibited Gsdmd expression and improved biomarkers of cardiac function in a rat model of DCM. On the whole, the findings of the present study indicate that BBR alleviates DCM by inhibiting miR­18a­3p­mediated Gsdmd activation; thus, BBR may be considered a potential therapeutic agent for the treatment of DCM.

[Effect of Lipoteichoic Acid Synthesis-Related Gene dltD on Acid Tolerance of Highly Cariogenic Strains of Streptococcus mutans].

Objective: To study the role and possible mechanism of dltD in the acid tolerance of Streptococcus mutans 593 (SM593), and to provide a theoretical basis for the ecological prevention and control of dental caries by constructing the dltD gene deletion strain of SM593 (SM593-ΔdltD). Methods: 1) SM593-Δ dltD was constructed by homologous recombination. 2) The growth curve of SM593 dltD and SM593-Δ dltD under different pH culture conditions was drawn by the automatic growth curve analyzer to compare their acid tolerance. Colony forming unit (CFU) at different time points was used to calculate the survival rate and to compare the acid tolerance response (ATR) of SM593 and SM593-Δ dltD. 3) Under different pH conditions, glycolysis experiments, proton permeability test and H +-ATPase activity test were conducted to make preliminary exploration into the mechanisms of how dltD gene deletion may affect acid tolerance. Results: 1) PCR and sequencing results showed that the SM593-Δ dltD was constructed successfully. 2) With decreasing pH value of the culture medium, the growth of SM593-Δ dltD slowed down. When the pH value of the culture medium was 5.0, SM593-Δ dltD was not allowed to grow, and its acid tolerance was lower than that of SM593. Compared with SM593, the ATR capability of SM593-Δ dltD was decreased. 3) SM593 dltD and SM593-Δ dltD did not show obvious difference in their glycolysis ability under different pH conditions. Compared with SM593 dltD, the proton permeability of SM593-Δ dltD under different pH conditions was increased significantly (P<0.05), and H +-ATPase activity decreased significantly (P<0.05). Conclusion: Compared with SM593 dltD, SM593-Δ dltD showed obvious decrease in acid tolerance, which may be caused by the significant increase in proton permeability and significant decrease in the H +-ATPase activity induced by the deletion of the dltD gene, hence reducing its ability to maintain intracellular pH homeostasis.

Comparison between minimally invasive deltoid-split and extended deltoid-split approach for proximal humeral fractures: a case-control study.

BACKGROUND: With the rapid aging of the population, the incidence of proximal humeral fracture (PHF) has increased. However, the optimal method for open reduction and internal fixation (ORIF) remains controversial. METHODS: We performed a retrospective analysis of patients with PHF who underwent locking plate internal fixation at our institution from January 2016 to December 2018. Patients were divided into two groups based on the surgical approach used: an expanded deltoid-split approach group (ORIF group) and minimally invasive deltoid-split approach group (minimally invasive percutaneous plate osteosynthesis, [MIPPO] group). The groups were compared in terms of demographic and perioperative characteristics, and clinical outcomes. RESULTS: A total of 115 cases of PHF were included in our study, of which 64 cases were treated using the minimally invasive deltoid-split approach and 51 using the extended deltoid-split approach. Fluoroscopy was performed significantly less frequently in the ORIF group and the surgical duration was shorter. However, the postoperative visual analogue scale (VAS) pain score and duration of postoperative hospital stay were significantly higher compared to the MIPPO group. Moreover, secondary loss was significantly less extensive in the ORIF group compared to the MIPPO group, while there was no significant group difference in fracture healing time, Constant shoulder score, or complications at the last follow-up visit. CONCLUSIONS: The clinical outcomes associated with both the minimally invasive and extended deltoid-split approaches were satisfactory. The data presented here suggest that the extended deltoid-split approach was superior to the minimally invasive deltoid-split approach in terms of operational time, fluoroscopy, and secondary loss of reduction, while the minimally invasive approach was superior in terms of postoperative pain and hospital stay. Accordingly, neither procedure can be considered definitively superior; the optimal surgical procedure for PHF can only be determined after full consideration of the situation and requirements of the individual patient.

Hidden blood loss and the influential factors after intramedullary nail fixation of extra-articular tibial fractures - a retrospective cohort study.

PURPOSE: There were few reports in the literature about the hidden blood loss (HBL) after intramedullary nail (IMN) fixation for extra-articular tibial fractures. Our purpose was to evaluate the amount of hidden blood loss after intramedullary nail fixation for extra-articular tibial fractures, meanwhile, identified the influential factors causing HBL. METHODS: From January 2015 to December 2017, 122 consecutive extra-articular tibial fracture patients fixed with IMN and 96 met all inclusion criteria for the chosen analysis. Preoperative hematocrit (Hct) levels on admission and postoperative Hct levels on the third day after surgery to calculate the amount of HBL. RESULTS: The mean HBL was 473.29 ± 102.75 ml after IMN fixation of extra-articular tibial fractures, the multiple regression analysis showed gender, surgical duration, and the diameter of the medullary cavity at the narrowest part of the tibia had an independent influence on HBL. CONCLUSIONS: A significant amount of postoperative HBL has occurred after IMN fixation of extra-articular tibial fractures. Surgeons should be aware that more HBL can be developed in patients who have male sex, small medullary cavity, and long-time surgical duration. HBL deserves attention to ensure patients' safety in the perioperative period of IMN fixation for extra-articular tibial fractures.

[Quality evaluation of Styrax based on quantitative determination of 7 chemical components using HPLC wavelength switching method and chemometrics].

In order to evaluate the quality of Styrax more comprehensively,this study attempted to establish an HPLC wavelength switching method to simultaneously determine the content of seven compounds in Styrax,and chemometrics were used to analyze the quality difference between different sources of Styrax,and finally establish a characteristic chromatogram of Styrax. The column was Agilent ZORBAX Extend C18( 4. 6 mm×250 mm,5 µm) with phase a mixture of acetonitrile-0. 1% phosphoric acid aqueous solution as the mobile phase in a gradient elution procedure; the detection wavelength was set as follows: 0-13. 5 min,194 nm( benzoic acid);13. 5-20. 5 min,278 nm( cinnamic acid); 20. 5-32 min,194 nm( benzyl benzoate,benzyl cinnamate,cinnamyl cinnamate,dehydroabietic acid); 32-55 min,241 nm( abietic acid). The methodological verification results showed that when the injection masses of benzoic acid,cinnamic acid,benzyl benzoate,benzyl cinnamate,cinnamyl cinnamate,dehydroabietic acid and abietic acid were0. 006 948-0. 694 8,0. 001 426-0. 142 6,0. 013 16-0. 658 0,0. 006 148-0. 614 8,0. 008 035-0. 803 5,0. 002 121-0. 212 1,and0. 010 172-1. 017 2 µg,respectively,there were good linear relationship between injection mass and peak area. The average recovery rates of seven compounds were in the range from 94. 34% to 105. 8%,and all RSD were less than 3. 0%( n = 6). The methodological verification results showed that the developed HPLC wavelength switching method has good accuracy and repeatability. The results of the sample analysis showed that the quality of Styrax from different sources was quite different. The chromatogram of Styrax reference material( S1) was used as the reference chromatogram to calculate the fingerprint similarity of each batch of samples. The results showed that the similarities of samples S2-S10 were 0. 952,0. 949,0. 981,0. 351,0. 751,0. 969,0. 979,0. 992 and 0. 971,respectively.The similarity values of other batches samples were satisfactory,except for sample S5 and S6,indicating that the quality difference among these samples is small. The similarity values of S11-S20 were 0. 060,0. 055,0. 054,0. 285,0. 092,0. 002,0. 044,0. 044,0. 044,and 0. 040,respectively. The results showed that compared with the sample S1,there was a large quality difference among S11-S20. Based on the chromatograms of S1-S10,the HPLC characteristic chromatograms of Styrax was established and the purpose is to give reference to other pharmaceutical researchers. The newly developed HPLC wavelength switching method have the advantages of simplicity,reproducibility and specificity,and the developed HPLC characteristic chromatograms provided a reference method for the overall quality evaluation of Styrax.

Modified citrus pectin inhibited bladder tumor growth through downregulation of galectin-3.

Modified citrus pectin (MCP) is a carbohydrate enriched complex, which has been implicated in cancer treatment and prevention. However, the effects of MCP on urinary bladder cancer (UBC) are unknown. In this study, MCP was first tested in T24 and J82 human UBC cells and showed the inhibition of cell viability by the sulforhodamine B (SRB) assay. The MCP-treated UBC cells exhibited G2/M phase arrest with the decrease of Cyclin B1 and phosphorylated Cdc2. Caspase-3 was also activated, leading to the cleavage of Caspase-3 and PARP. We further explored the possible molecular mechanisms upon MCP treatment in UBC cells. Reduction of galectin-3 was observed and followed with the inactivation of Akt signaling pathway. Of note, galectin-3 knockdown by RNA interference recapitulated the MCP-mediated anti-proliferation, cell cycle arrest and apoptosis. Moreover, oral administration of MCP to the T24 xenograft-bearing nude mice inhibited the tumor growth significantly (P < 0.05). Quantification analysis of immunohistochemistry staining for Ki67 and cleaved Caspase-3 confirmed the decrease of proliferation index (P < 0.05) and the increase of apoptosis index (P < 0.01) in 700 mg/kg MCP-fed UBC xenografts. Using the information from TCGA database, we revealed that the overexpression of galectin-3 was associated with high tumor grade with lymph node metastasis, poor overall survival in UBC patients. Considering the remarkable inhibitory effects of MCP on UBC cell proliferation and survival in vitro and in vivo mainly through galectin-3, which is upregulated in UBCs, MCP may become an attractive agent, as a natural dietary fiber, for prevention and therapy of UBCs.

The complete mitochondrial genome of the Lethrinops lethrinus.

In the present study, for the first time, we reported the complete mitochondrial genome of Lethrinops lethrinus, which is 16,582 bp in length, including 13 protein-coding genes, 2 ribosomal RNA, 22 transfer RNA genes and a control zone. Moreover, its GC content is 45.99% (27.42% A, 26.59% T, 30.10% C, and 15.88% G), similar to that of Alticorpus geoffreyi (GC content of 45.82%). We further made the phylogenetic tree on the complete mitochondrial genomes of the above two species and other 12 closely related species to show their phylogenic relationship. The above results would facilitate our understanding of the evolution of Cichlidae mitochondrial genome.

Butylphthalide Suppresses Neuronal Cells Apoptosis and Inhibits JNK-Caspase3 Signaling Pathway After Brain Ischemia /Reperfusion in Rats.

Although Butylphthalide (BP) has protective effects that reduce ischemia-induced brain damage and neuronal cell death, little is known about the precise mechanisms occurring during cerebral ischemia/reperfusion (I/R). Therefore, the aim of this study was to investigate the neuroprotective mechanisms of BP against ischemic brain injury induced by cerebral I/R through inhibition of the c-Jun N-terminal kinase (JNK)-Caspase3 signaling pathway. BP in distilled non-genetically modified Soybean oil was administered intragastrically three times a day at a dosage of 15 mg/(kg day) beginning at 20 min after I/R in Sprague-Dawley rats. Immunohistochemical staining and Western blotting were performed to examine the expression of related proteins, and TUNEL-staining was used to detect the percentage of neuronal apoptosis in the hippocampal CA1 region. The results showed that BP could significantly protect neurons against cerebral I/R-induced damage. Furthermore, the expression of p-JNK, p-Bcl2, p-c-Jun, FasL, and cleaved-caspase3 was also decreased in the rats treated with BP. In summary, our results imply that BP could remarkably improve the survival of CA1 pyramidal neurons in I/R-induced brain injury and inhibit the JNK-Caspase3 signaling pathway.

The complete mitochondrial genome of the Fossorochromis rostratus.

The Fossorochromis rostratus is a species of cichlid endemic to Lake Malawi in East Africa. In this study, we first reported the complete mitochondrial genome of F. rostratus. The whole mitochondrial genome is 16 581 bp in length, which contains 13 protein-coding genes, 22 transfer RNA genes, and two ribosomal RNA genes. The GC content of this mitochondrial genome is 45.96% (27.47% A, 26.57%T, 30.12% C, and 15.84% G), similar to Astatotilapia calliptera (the GC content of 45.90%). We constructed a phylogenetic tree on the complete mitochondrial genomes of these two species and other 10 closely related species to show their phylogenic relationship. The complete mitochondrial genome of F. rostratus and its phylogenic relationship with other related species would facilitate our understanding of the evolution of Cichlidae mitochondrial genome.

The complete mitochondrial genome of the Alticorpus geoffreyi.

The present study reported the complete mitochondrial genome of Alticorpus geoffreyi for the first time. The mitochondrial genome of A. geoffreyi possesses 16,578 bp in length, involving 22 transfer RNA genes, 2 ribosomal RNA, 13 protein-coding genes and one control region. In addition, its GC content is 45.82% that is similar to that of Astatotilapia calliptera (the GC content of 45.90%). Based on the complete mitochondrial genomes of 14 closely related species, the phylogenetic tree was further made to show their phylogenic relationship. The results will serve as a useful dataset for studying the evolution of Cichlidae mitochondrial genome.

The complete mitochondrial genome of the Copadichromis mloto.

The present study reported the complete mitochondrial genome of Copadichromis mloto for the first time. The mitochondrial genome of C. mloto possesses 16,583bp in length, involving 22 transfer RNA genes, 2 ribosomal RNA, 13 protein-coding genes and a control region. In addition, its GC content is 45.94% that is similar to that of C. virginalis (the GC content of 45.74%). Based on the complete mitochondrial genomes of 13 closely related species, the phylogenetic tree was further made to show their phylogenic relationship. The results will serve as a useful dataset for studying the evolution of Cichlidae mitochondrial genome.

Point application with Angong Niuhuang sticker protects hippocampal and cortical neurons in rats with cerebral ischemia.

Angong Niuhuang pill, a Chinese materia medica preparation, can improve neurological functions after acute ischemic stroke. Because of its inconvenient application and toxic components (Cinnabaris and Realgar), we used transdermal enhancers to deliver Angong Niuhuang pill by modern technology, which expanded the safe dose range and clinical indications. In this study, Angong Niuhuang stickers administered at different point application doses (1.35, 2.7, and 5.4 g/kg) were administered to the Dazhui (DU14), Qihai (RN6) and Mingmen (DU4) of rats with chronic cerebral ischemia, for 4 weeks. The Morris water maze was used to determine the learning and memory ability of rats. Hematoxylin-eosin staining and Nissl staining were used to observe neuronal damage of the cortex and hippocampal CA1 region in rats with chronic cerebral ischemia. The middle- and high-dose point application of Angong Niuhuang stickers attenuated neuronal damage in the cortex and hippocampal CA1 region, and improved the memory of rats with chronic cerebral ischemia with an efficacy similar to interventions by electroacupuncture at Dazhui (DU14), Qihai (RN6) and Mingmen (DU4). Our experimental findings indicate that point application with Angong Niuhuang stickers can improve cognitive function after chronic cerebral ischemia in rats and is neuroprotective with an equivalent efficacy to acupuncture.

[Preliminary study on general regulations for processing of different processing procedures of prepared slices of Chinese crude drugs in China].

General regulations for the processing are the important part of processing procedures of prepared slices of Chinese crude drugs. It has an important significance on enhancing the operability of actual production, regulating production of prepared slices of Chinese crude drugs, improving quality and establishing drug safety. The article could provides suggestions and reference for future compilation work on "National processing procedures of prepared slices of Chinese crude drugs" by comparative analysis and summary on general regulations for the processing of different processing procedures of prepared slices of Chinese crude drugs in China.

[Protein expression by planktonic and biofilm cells of clinical isolations of Streptococcus mutans].

OBJECTIVE: To investigate the changes in the surface-associated protein expression of planktonic cells and biofilm cells of clinical isolations of Streptococcus mutans (Sm). METHODS: The proteins were extracted by the method of Homer from the nonadhered planktonic and the adhered biofilm cells and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis followed by image analysis. Proteins were identified by matrix-assisted laser desorption time of flight mass spectrometry and computer-assisted protein sequence analysis. RESULTS: Image analysis revealed that 30% - 31% of the protein spots in biofilm was modulated. A total of 238 proteins changed 1.3-fold or greater in biofilm cells compared to planktonic cells in Sm18, with 5 only expressed in biofilm cells and 5 not expressed in biofilm cells. A total of 279 proteins changed 1.3-fold or greater in biofilm cells compared to planktonic cells in Sm593, with 12 only expressed in biofilm cells and 2 not expressed in biofilm cells. Two clinical isolations in biofilm cells had three identical protein expressions whose functions were associated with biosynthetic processes. CONCLUSIONS: The two clinical isolations in biofilm status have high expression of some special proteins, which are presumed to be key proteins essential for formation of biofilm. The difference of protein expression in biofilm cells of the two clinical isolations may represent their distinct characters.

[Effect of the self-etching adhesives system on human pulp fibroblast].

OBJECTIVE: To compare and evaluate the biocompatibility of three kinds of dentin bonding agents Xeno III (XO), Adper Prompt (AP), Single bond2 (SB) through cell culture in vitro. METHODS: Three kinds of dentin bonding agents (XO, AP, SB) were applied on the surface of the dental slices which were 5.0 mm in diameter and 0.5 mm in depth. By immersing the slices into the DMEM culture medium, the maceration extracts were obtained. Normal dental pulps of teenagers were collected and human pulp fibroblast was cultured using tissue explant method. The fifth generation pulp cells were exposed to culture medium containing different concentrations of maceration extracts (100.0%, 50.0%, 25.0%, 12.5%) for 24, 72, 120 h. At last, MTT method was used to evaluate the cytotoxicity of the dentin bonding agents on human pulp fibroblast. RESULTS: The results showed that all three kinds of dentin bonding systems had cytotoxicity to human pulp fibroblast in different degree in vitro. The cytotoxicity of XO and AP was less than SB. The difference was statistically significant (P<0.05). CONCLUSION: The results of cell culture in vitro indicated that total-etching adhesives system has more irritation to pulp than self-etching adhesives system.

[The safety and efficiency of fast track surgery in gastric cancer patients undergoing D2 gastrectomy].

OBJECTIVE: To investigate the safety and efficacy of fast track surgery (FTS) management in gastric cancer undergoing D2 gastrectomy. METHODS: Eighty gastric cancer patients undergoing D2 gastrectomy were recruited prospectively. Patients were assigned to receive FTS management (n = 40) or conventional perioperative care (n = 40). The FTS care included shorten preoperative fasting time, no nasogastric decompressing tubes and abdominal drainage placed, early postoperative oral feeding, multimodal analgesia, and early mobilisation. The length of postoperative hospital stay, medical cost, nutritional status, gut function, and postoperative complications in the two groups were recorded and compared. RESULTS: FTS group was associated with a significantly shorter postoperative hospital stay compared with conventional care group [(5.6 +/- 1.3) d vs. (9.4 +/- 1.9) d, P < 0.05]. Medical cost was less [(18 620 +/- 2360) Yuan vs. (20 370 +/- 2440) Yuan, P < 0.05] and duration of intravenous infusion [(3.5 +/- 1.4) d vs. (5.8 +/- 1.9) d, P < 0.05] was also shorter. First passage of flatus was earlier in FTS group than in conventional care group [(4.3 +/- 0.4) d vs. (5.5 +/- 0.9) d, P < 0.05]. Loss of body weight in the postoperative period was less in FTS group [(3.2 +/- 0.8) kg vs. (4.3 +/- 1.6) kg, P < 0.05]. There was no difference in morbidity or mortality between the two groups. CONCLUSION: FTS in D2 gastrectomy is safe and efficient, and it can shorten postoperative hospital stay and hasten return of gut function.

[Technique of detection of hepatitis C core antigen used in safety blood transfusion].

This study was purposed to investigate the feasibility to screen donor with HCV infection by means of HCV-cAg ELISA. The first and repeat assays were performed for detection of serum anti-HCV in 8677 donor's serum specimens from January 2003 to December 2005. All serum anti-HCV specimens with positive anti-HCV from first and repeat assays were finally identified by using HCV-cAg ELISA and HCV RT-PCR methods. The results showed that only 5 serum specimens were positive anti-HCV by HCV-cAg ELISA identification in 29 specimens including 15 specimens with positive ant-HCV in first assays and 14 specimens with positive anti-HCV in repeat assays, the positive rate detected by HCV cAg ELISA was 17.24%. 5 serum specimens were positive anti-HCV by HCV RT-PCR detection also in 29 specimens mentioned above, the positive rate detected by HCV RT-PCR was 17.24% too. It is concluded that sensitivity of HCVcAg ELISA is similar to HCV RT-PCR and may be useful for the early diagnosis of hepatitic C or used as a reliable method to screen donor with HCV infection in blood transfusion medicine.

[Evaluation of the in vitro cariogenic potential of Streptococcus mutans (serotype C) strains isolated from caries-free and -active people: the ability of acidogenicity].

OBJECTIVE: To investigate the ability of acidogenicity of S. mutans (serotype C) strains isolated from the people with different caries experience. METHODS: Same density solutions of all isolated S. mutans were made and cultured in different pH TPPE liquid for the same period of time. Terminal pH of the solutions was measured. The values of delta pH were compared. RESULTS: Significant difference of acidogenicity was observed between the strains of different genotypes isolated from the same person. The ratio of high acidogenicity isolates harbored in caries-active people was greater than that of caries-free people; a significant difference was noted (P<0.05). CONCLUSION: The high cariogenicity of isolated S. mutans strains of caries-active people shows a close relationship with the high acidogenicity of the isolated S. mutans (serotype C) strains.