In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics. Similarities and variations of components present significant analytical challenges. A two-dimensional (2D) liquid chromatography-mass spectrometr (LC-MS) method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium (CMS). A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated. For efficient and high-accuracy screening of CMS, a targeted method based on a self-constructed high resolution (HR) mass spectrum database of CMS components was established. The database was built based on the commercial MassHunter Personal Compound Database and Library (PCDL) software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening. On this basis, the unknown peaks in the CMS chromatograms were deduced and assigned. The molecular formula, group composition, and origins of a total of 99 compounds, of which the combined area percentage accounted for more than 95% of CMS components, were deduced by this 2D-LC-MS method combined with the MassHunter PCDL. This profiling method was highly efficient and could distinguish hundreds of components within 3 h, providing reliable results for quality control of this kind of complex drugs.

Objective To analyze the literature related to diphenidol tablets poisoning,the characteristics of poisoning were summarized to provide reference for controlling the suicidal abuse risk of diphenidol tablets.Methods The global literature on suicide,overdose,poisoning,shock,and death related to difenidol published from January 1,2011 to December 31,2022 was analyzed,including gender,age,dosage,cardiac(blood)concentration,poisoning symptoms,etc.Results Young women were the majority of people with poisoning.The highest proportion of the age group is 11 to 30 years group.Patients who take medication doses greater than 3 000 mg may have a higher risk of death;patients with a heart(blood)concentration greater than 6 μg·mL-1 may have a higher risk of death.Malignant arrhythmia,consciousness disorders,coma,and apnea are common serious adverse events during poisoning.Conclusion It is recommended that the drug regulatory authorities should require the Listing permit holder of difenidol tablets to add the risk and symptoms of poisoning into the instructions.It is suggested that restricting individual consumers from purchasing large amounts of difenidol tablets in the short term.It is recommended that canceling the high-dose sales packaging of difenidol tablets.It is suggested that converting difenidol tablets into prescription drugs,even consider canceling the registration certificate of difenidol tablets.

The present study aimed to investigate the protective role of Shaofu Zhuyu Decoction(SFZY) against endometriosis fibrosis in mice, and decipher the underlying mechanism through the phosphatase and tensin homolog deleted on chromosome ten(PTEN)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway. Eighty-five BALB/c female mice were randomly assigned into a blank group, a model group, high-, medium, and low-dose SFZY(SFZY-H, SFZY-M, and SFZY-L, respectively) groups, and a gestrinone suspension(YT) group. The model of endometriosis was induced by intraperitoneal injection of uterine fragments. The mice in different groups were administrated with corresponding groups by gavage 14 days after modeling, and the blank group and model group with equal volume of distilled water by gavage. The treatment lasted for 14 days. The body weight, paw withdrawal latency caused by heat stimuli, and total weight of dissected ectopic focus were compared between different groups. The pathological changes of the ectopic tissue were observed via hematoxylin-eosin(HE) and Masson staining. Real-time PCR was employed to measure the mRNA levels of α-smooth muscle actin(α-SMA) and collagen type Ⅰ(collagen-Ⅰ) in the ectopic tissue. The protein levels of PTEN, Akt, mTOR, p-Akt, and p-mTOR in the ectopic tissue were determined by Western blot. Compared with the blank group, the modeling first decreased and then increased the body weight of mice, increased the total weight of ectopic focus, and shortened the paw withdrawal latency. Compared with the model group, SFZY and YT increased the body weight, prolonged the paw withdrawal latency, and decreased the weight of ectopic focus. Furthermore, the drug administration, especially SFZY-H and YT(P<0.01), recovered the pathological and reduced the area of collagen deposition. Compared with the blank group, the modeling up-regulated the mRNA levels of α-SMA and collagen-Ⅰ in the ectopic focus, and such up-regulation was attenuated after drug intervention, especially in the SFZY-H and YT groups(P<0.05,P<0.01). Compared with the blank group, the modeling down-regulated the protein level of PTEN and up-regulated the protein levels of Akt, mTOR, p-Akt, and p-mTOR(P<0.01, P<0.001). Drug administration, especially SFZY-H and YT, restored such changes(P<0.01). SFZY may significantly attenuate the focal fibrosis in the mouse model of endometriosis by regulating the PTEN/Akt/mTOR signaling pathway.
Female ; Animals ; Mice ; Humans ; Proto-Oncogene Proteins c-akt/genetics* ; Choristoma ; Endometriosis/genetics* ; TOR Serine-Threonine Kinases/genetics* ; RNA, Messenger ; Signal Transduction ; Body Weight ; Mammals ; PTEN Phosphohydrolase/genetics*

This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.
Humans ; Carcinoma, Hepatocellular/genetics* ; Proto-Oncogene Proteins c-akt/metabolism* ; Caspase 3/metabolism* ; Liver Neoplasms/genetics* ; Molecular Docking Simulation ; Sincalide/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Hep G2 Cells ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.
Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis ; Autophagy ; Cell Proliferation ; Cell Line, Tumor ; STAT3 Transcription Factor/metabolism*

Objective To analyze the status of quality indicators(QI) on specimen acceptability and establish preliminary qual ity specification.Methods Web based External Quality Assessment system was used to collect data of laboratories partici pated in "Medical quality control indicators in clinical laboratory" from 2015 to 2017,including once in 2015 and 2017 and twice in 2016.Rate and sigma scales were used to evaluate incorrect sample type,incorrect sample container,incorrect fill level and anticoagulant sample clotted.The 25th percentile (P25) and 75th percentile (P75) of the distribution of each QI were employed to establish the high,medium and low specification.Results 5 346,7 593,5 950 and 6 874 laboratories sub mitted the survey results respectively.The P50 of biochemistry (except incorrect fill level),immunology and microbiology reach to 6σ.The P50 of clinical laboratory is 4 to 6σ except for incorrect sample container.There is no significant change of the continuous survey results.Based on results in 2017 to establish the quality specification,the P25 and P75 of the four QIs is 0 and 0.084 4 ％,0 and 0.047 6 ％,0 and 0.114 2 ％,0 and 0.078 4 ％,respectively.Conclusion According to the results of the survey,most laboratories had a faire performance in biochemistry,immunology and microbiology,and clinical laboratory needs to be strengthened.Laboratories should strengthen the laboratory information system construction to ensure the actual and reliable data collection,and make a long time monitoring to achieve a better quality.

OBJECTIVE: To observe cardiac ultrastructure and the expression of heat shock protein 70 (HSP70) and hypoxia inducible factor-lα (HIF-lα) in electric shock death rats and to explore the application of these indexes as the basis of medical identification in electric shock death. METHODS: Seventy-two SD rats were randomly divided into electric shock death group, postmortem electric shock group and the control group. The changes of myocardial ultrastructure were observed by transmission electron microscope, and the expressions of myocardial HSP70 and HIF-1α were observed by immunohistochemical technology. RESULTS: Myocardial myofibril fracture, mitochondrial cristae and membrane dissolution, and disordered arrangement of Z lines and M lines were observed in electric shock rats. HSP70 and HIF-lα were strong positive expressions in the electric shock death group, significantly compared with the control and postmortem electric shock groups (P < 0.05). CONCLUSION The expressions of HSP70 and HIF-lα were obviously increased in electric shock death group, which may be used as the diagnostic indicator of electric shock death.
Animals ; Death ; HSP70 Heat-Shock Proteins/metabolism* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Myocardium/pathology* ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the efficacy and toxicity of paclitaxel with low-dose cisplatin and 5-fluorouracil continuous infusion in the treatment of advanced gastric carcinoma.

METHODSThe patients were treated with paclitaxel liposome 60 mg/m(2) i.v. gtt on d1, 8, 15, DDP 15 mg×m(-2)×d(-1) by i.v. gtt on d1-5, 5-Fu 500 mg×m(-2)×d(-1) by civ for 120 h, administered every 21 days.

RESULTSOut of the whole group, 3 cases achieved CR, 29 cases achieved PR with an ORR of 54.2% and median TTP of 7.1 months. Out of 40 cases in the primary treatment, 3 cases achieved CR, 22 cases achieved PR with an ORR of 62.5% and median TTP of 7.6 months. Out of 20 evaluable retreated cases, no case achieved CR, 7 cases achieved PR with an ORR of 36.8% and median TTP 6.3 months. The main toxicities were hematological toxicities, nausea and vomiting of grade I-II.

CONCLUSIONThe combination regimen of paclitaxel, low-dose cisplatin and 5-fluorouracil is effective and well tolerated for patients with advanced gastric carcinoma, especially for primary treatment cases. It is worthy of further study.

Adenocarcinoma ; drug therapy ; pathology ; secondary ; surgery ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; adverse effects ; therapeutic use ; Cisplatin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Infusions, Intravenous ; Leukopenia ; chemically induced ; Liposomes ; Liver Neoplasms ; drug therapy ; secondary ; Male ; Middle Aged ; Nausea ; chemically induced ; Neoplasm Staging ; Paclitaxel ; administration & dosage ; adverse effects ; Remission Induction ; Stomach Neoplasms ; drug therapy ; pathology ; surgery ; Survival Rate ; Vomiting ; chemically induced ; Young Adult

OBJECTIVETo evaluate the effects of mixed grafting of allogeneic PADM and autologous STS on wound healing of full-thickness defect in rats.

METHODSFull-thickness defects with size of 6 cm x 4 cm were produced on the back of 12 SD rats, and they were divided into E group (n = 6) and C group (n = 6) according to the random number table. The wounds in E group were grafted with a mix of allogeneic PADM (expansion rate 10: 5) and autologous STS with thickness of 0.2 mm, while those in C group were grafted with autologous STS in the same thickness. The wound healing rate, survival rate, contraction rate, and expansion rate of transplanted skin were observed at post operation week (POW) 2, 3, 4, 6, 8, 12, 20. Tissue samples form wounds and surrounding normal skin were harvested at POW 20 for histopathological observation as follows. The structure of collagen fiber bundle was observed by HE staining, the diameter and gap rate of collagen fiber bundle were also measured. The distribution of type I and III collagen was observed by sircus red staining, and the contents of type I, III collagen and their ratio were also examined. Data were processed with independent samples t test, Levene test, and t' test.

RESULTSSurvival rate of transplanted skin in E group at POW 2 [(76.1 +/- 13.1)%] was obviously lower than that in C group [(94.5 +/- 1.3)%, t' = 3.440, P = 0.018]. Contraction rate of transplanted skin in E, C groups at POW 3 showed significant difference [(34 +/- 8)% vs. (16 +/- 12)%, t = -3.211, P = 0.009]. Compared with those in peri-wound normal skin, collagen fiber bundles in C group showed signs of homogenization, and collagen fibers were thin with irregular arrangement. Collagen fiber structure and arrangement of composite skin in E group were similar to those surrounding normal skin with incomplete degradation of PADM. Diameter of collagen fiber bundle [(9.6 +/- 0.8) microm], gap rate between collagen bundle [(24 +/- 5)%], content of type I collagen [(80.2 +/- 5.4)%] and the ratio of type I to type III collagen (4.3 +/- 1.2) in E group were all increased as compared with those in C group [(7.3 +/- 1.4) microm (t = -3.562, P = 0.005), (17 +/- 4)% (t = -2.760, P = 0.020), (68.1 +/- 8.4)% (t = -2.981, P = 0.014), 2.3 +/- 1.0 (t = -3.204, P = 0.009)], while content of type III collagen [(19.8 +/- 5.4)%] in E group was lower than that in C group [(32.0 +/- 8.4)%, t = 2. 981, P = 0.014]. Above-mentioned indexes of collagen in wound of E group were similar to those of normal skin surrounding the wound.

CONCLUSIONSAllogeneic PADM used as dermal regeneration template is beneficial in improving collagen fiber bundle structure in dermis layer of rats with full-thickness skin wounds when repaired with autologous STS, and it accelerates maturation of regenerative dermal tissue.

Animals ; Burns ; surgery ; Dermatologic Surgical Procedures ; Dermis ; cytology ; transplantation ; Extracellular Matrix ; transplantation ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; injuries ; Skin Transplantation ; Skin, Artificial ; Transplantation, Autologous ; Treatment Outcome ; Wound Healing

Objective To investigate the effect of 131I on apoptosis of thyrocytes in patients with Graves disease. Methods Forty-seven patients with Graves disease were divided into two groups, two week group (G2w) and four week group (G4w). All patients underwent thyoid needle biopsy before 131I treatment and the repeated biopsy at two weeks (G2w) or four weeks (G4w) after 131I treatment. The positive units of pro-apoptotic proteins (Fas, FasL) and anti-apoptotic protein (Bcl-2) were studied with immunohistochemistry staining. The differences of the two groups were compared with t-test. Liner correlation analysis was applied to study the correlation between 131I dose and apoptosis-related proteins and that between serum sTSH after 131I treatment and apoptosis-related proteins. Results Fas, FasL and Bcl-2 expression (positive units) were significantly increased in both groups after 131I treatment, G2w :22.84 ± 9.31 vs 16.20 ± 6.75,21.13±6.29vs 14.56±4.06, 21.69±7.83 vs 15.22 ±5.94, t= -3.08, -3.73, -4.05 (allP<0.05); G4w:21.69 ±4.52 vs 15.83 ±5.03, 19. 11 ±3.75 vs 14.02 ±4.98, 19.06 ±3.44 vs 16.63 ±4. 73, t = - 5.26, - 5.00, - 2.41 (all P<0.05). However, no statistical differences were found between G2w and G4w (t = 0. 53, 0. 82, 1.46, all P > 0.05). Significant correlation was found between 131I 0. 727, rFasL = 0. 763 (both P<0.05)), but not between the dose and Bcl-2, rBcl-2 = - 0. 094, 0. 102(both P > 0.05). There were significant correlation between serum sTSH three months after 131I treatment and apoptosis-related proteins, rFas = 0.433, rFasL = 0. 601, rBcln2 = - 0. 397, (all P<0. 05). Conclusions 131I can induce thyrocytes to express the pro-apoptotic proteins in patients with Graves disease.