Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Objective To investigate the effects of parecoxib sodium injection combined with dexmedetomidine hydrochloride injection on postoperative cognitive function and stress response in patients with osteoporotic compression fractures.Methods The patients with osteoporotic compression fractures were divided into treatment group and control group according to the treatment plan.The control group was given intravenous injection of dexmedetomidine hydrochloride injection 0.2 μg·kg-1load dose,then micro pump injection 0.2 μg·kg-1·min-1 maintenance dose,until 30 min before the end of the operation;patients in the treatment group were intravenously injected with parecoxib sodium injection 20 mg before local anesthesia and 30 min before the end of operation on the basis of the control group.The pain,sedation,hemodynamics[mean arterial pressure(MAP),heart rate(HR)],cognitive function and safety evaluation were compared between the two groups before operation(T0),2 h after operation(T1),6 h after operation(T2),12 h after operation(T3)and 24 h after operation(T4).Results There were 39 cases in the treatment group and 41 cases in the control group.Visual analogue scale(VAS)scores in treatment group and control group were(3.09±0.55)and(3.41±0.62)scores at T1;VAS scores were(3.02±0.57)and(3.35±0.48)scores at T2;VAS scores were(2.64±0.44)and(2.90±0.46)scores at T3;VAS scores were(2.02±0.41)and(2.35±0.47)scores at T4;MMSE scores were(25.28±1.57)and(24.33±1.42)scores at T2;MMSE scores were(28.16±1.01)and(27.25±0.89)scores at T4;MoCA scores were(24.63±1.60)and(23.59±1.25)scores at T2;MoCA scores were(27.20±0.97)and(26.48±0.83)scores at T4.There were statistically significant differences in the above indexes between the treatment group and the control group(all P＜0.05).Adverse drug reactions in the treatment group included bradycardia,hypotension,nausea vomiting and hypokalemia;adverse drug reactions in the control group included bradycardia,hypotension and nausea vomiting.The total incidence rates of adverse drug reactions were 12.82％and 9.76％,without statistically significant difference(P＞0.05).Conclusion Compared with using dexmedetomidine alone,parecoxib sodium combined with dexmedetomidine is beneficial for relieving postoperative pain in patients with osteoporotic compression fractures,improving postoperative cognitive function.

OBJECTIVE: To observe the effects of electro-scalp acupuncture （ESA） on the expression of microglial markers CD206 and CD32, as well as interleukin (IL)-6, IL-1β, and IL-10 in the ischemic cortex of rats with ischemic stroke, and to explore the mechanisms of ESA on alleviating inflammatory damage of ischemic stroke. METHODS: Sixty 7-week-old male SD rats were randomly selected, with 15 rats assigned to a sham surgery group. The remaining rats were treated with suture method to establish rat model of middle cerebral artery occlusion (MCAO). The rats with successful model were randomly divided into a model group, a VitD₃ group, and an ESA group, with 15 rats in each group. In the ESA group, ESA was performed bilaterally at the "top-temporal anterior oblique line" with disperse-dense wave, a frequency of 2 Hz/100 Hz, and an intensity of 1 mA. Each session lasted for 30 min, once daily, for a total of 7 days. The VitD₃ group were treated with intragastric administration of 1,25-dihydroxyvitamin D₃ (1,25-VitD₃) solution (3 ng/100 g), once daily for 7 days. The neurological deficit scores and neurobehavioral scores were assessed before and after the intervention. After the intervention, the brain infarct volume was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Immunofluorescence double staining was performed to detect the protein expression of CD32 and CD206 in the ischemic cortex. Western blot analysis was conducted to measure the protein expression of IL-6, IL-1β, and IL-10 in the ischemic cortex. RESULTS: Compared with the sham surgery group, the model group showed increased neurological deficit scores and neurobehavioral scores (P<0.01), increased brain infarct volume (P<0.01), increased protein expression of CD32, IL-6, and IL-1β in the ischemic cortex (P<0.01), and decreased protein expression of CD206 and IL-10 in the ischemic cortex (P<0.01). Compared with the model group, both the ESA group and the VitD₃ group showed decreased neurological deficit scores and neurobehavioral scores (P<0.01), reduced brain infarct volume (P<0.01), decreased protein expression of CD32, IL-6, and IL-1β in the ischemic cortex (P<0.01), and increased protein expression of CD206 and IL-10 in the ischemic cortex (P<0.01). Compared with the VitD₃ group, the ESA group had lower neurological deficit score (P<0.05), larger brain infarct volume (P< 0.05), and lower protein expression of CD32, CD206, IL-1β, and IL-10 in the ischemic cortex (P<0.01, P<0.05). CONCLUSION ESA could improve neurological function in MCAO rats, and its mechanism may be related to promoting microglial M1-to-M2 polarization and alleviating inflammatory damage.
Male ; Animals ; Rats ; Rats, Sprague-Dawley ; Ischemic Stroke ; Interleukin-10 ; Interleukin-6/genetics* ; Microglia ; Scalp ; Acupuncture Therapy ; Vitamins ; Infarction, Middle Cerebral Artery

1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.

Objective: To explore the clinical effects of proximal ulnar artery perforator flap combined with iliac bone graft in the reconstruction of subtotal thumb or finger defects. Methods: A retrospective observational study was conducted. From August 2016 to August 2019, 7 patients with thumb or finger defects caused by mechanical damage who met the inclusion criteria were admitted to Ruihua Affiliated Hospital of Soochow University, including 6 males and 1 female, aged 46 to 58 years. Their length of fingers was repaired with iliac bone, with length of 2.0 to 3.0 cm. After the bone graft, the skin defect area of the affected finger ranged from 2.8 cm×2.2 cm to 6.0 cm×3.2 cm. Then the free proximal ulnar artery perforator flap with area of 3.0 cm×2.4 cm to 6.5 cm×3.5 cm was used to cover the wounds. The wounds in donor sites of iliac crest and flap were directly sutured. The survival of flap in one week post surgery and the donor site wound healing in 2 weeks post surgery were observed, respectively. During the follow-up, the appearance and sensory function of the affected finger, bone healing, and scar hypertrophy of wound in the donor site were observed and evaluated. At the last follow-up, the functional recovery of the affected finger was evaluated with trial standard for the evaluation of functions of the upper limbs of the Hand Surgery Society of Chinese Medical Association. Results: In one week post surgery, all the flaps survived. In 2 weeks post surgery, the iliac bone and the wounds in forearm donor site healed. During the follow-up of 5 to 13 months, the flap was good in appearance, without obvious pigmentation; the sensory recovery reached level S₂ in 5 patients and S₀ in 2 patients; all the grafted iliac bones were bony union without obvious resorption; the wounds in donor site healed well, with only mild scar formation. At the last follow-up, the shape of the reconstructed finger was close to the healthy finger, and the functional evaluation results were excellent in 3 cases and good in 4 cases. Conclusions: The use of proximal ulnar artery perforator flap combined with iliac bone graft to reconstruct subtotal thumb or finger can partially restore part of the appearance and function, with less damage to the donor site. It is a good choice for patients who have low expectations of appearance and function for the reconstructed finger.
Male ; Humans ; Female ; Soft Tissue Injuries/surgery* ; Perforator Flap/transplantation* ; Skin Transplantation/methods* ; Thumb/surgery* ; Plastic Surgery Procedures ; Ulnar Artery/surgery* ; Cicatrix/surgery* ; Ilium/surgery* ; Treatment Outcome

Objective:To explore the role of macrophage polarization on pericyte-to-myofibroblast transition and renal allograft fibrosis after kidney transplantation(KT).Methods:Allograft tissues were harvestedfrom recipients with chronic allograft dysfunction(CGD)and normal kidney tissues.The expression and distribution of M1/M2 macrophages in kidney tissues were detected by routine and immunofluorescent staining; mRNA of CD68, CD206 and iNOS detected by polymerase chain reaction(PCR); Murine vascular pericytes subjected to TGF-β1 in vitro and the expressions of α-SMA and PDGFR-β in perivascular cells detected by immunoblotting and cellular fluorescence; The co-culturing models of vascular pericytes and M1/M2 macrophages were constructed.The expressions of α-SMA and PDGFR-β in pericytes were detected by immunoblotting, cellular fluorescence and PCR.Results:A marked infiltration of CD68+ iNOS+ M1 macrophages was present in allograft tissues of recipients with CGD while no obvious infiltration of CD68 + CD206 + was observed.The mRNA levels of CD68, iNOS and CD206 were significantly higher in CGD group than those in control group( P<0.05); In CGD allograft tissues, protein expressions of α-SMA and PDGFR-β spiked markedly( P<0.05)while cells with double staining of α-SMA and PDGFR-β were markedly infiltrated in interstitial area of CGD allograft.TGF-β1 could induce a marked elevation of PMT-related markers in a time-dependent manner( P<0.05); Immunoblotting and cellukar fluorescence indicated that M1 macrophages could promote the elevations of α-SMA and PDGFR-β in pericytes in vitro while M2 macrophages showed no effect on pericyte-to-myofibroblast transition in pericytes. Conclusions:M1 macrophage polarization may promote the formation of renal allograft interstitial fibrosis through promoting PMT.

Objective:To investigate the correlation between collateral circulation and infarct pattern and outcome in acute ischemic stroke patients with anterior circulation intracranial atherosclerosis.Methods:Acute ischemic stroke patients with anterior circulation intracranial atherosclerotic severe stenosis or occlusion admitted to the Department of Neurology, the First Affiliated Hospital of Xinxiang Medical College from September 2018 to March 2020 were included prospectively. According to diffusion-weighted imaging, the infarct patterns were divided into perforator pattern, territorial pattern, watershed pattern, and mixed pattern. At 90 d after onset, the modified Rankin Scale was used to evaluate the outcome. 0-2 was defined as good outcome, and >2 was defined as poor outcome. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 89 patients were enrolled, 50 (56.2%) had good collateral circulation and 39 (43.8%) had poor collateral circulation. The distribution patterns of infarct: 22 patients (24.7%) were perforator pattern, 26 (29.2%) were territorial pattern, 17 (19.1%) were watershed pattern, and 24 (30.0%) were mixed pattern. The proportion of patients with good collateral circulation was 81.8%, 65.4%, 29.4% and 41.7%, respectively in the perforator pattern group, territorial pattern group, watershed pattern group, and mixed pattern group. Good collateral circulation was more common in the perforator pattern group, and poor collateral circulation was more common in the watershed pattern group. At 90 d after onset, 53 patients (59.6%) had a good outcome and 36 (40.4%) had a poor outcome. The baseline homocysteine level in the good outcome group was significantly lower than that in the poor outcome group (17.91±4.62 μmol/L vs. 20.35±4.67 μmol/L; t=2.436, P=0.017), and the proportion of patients with good collateral circulation was significantly higher than that of patients with poor outcome (73.6% vs. 30.6%; χ2=16.124, P<0.001). Multivariate logistic regression analysis showed that higher homocysteine level was an independent risk factor for poor outcome (odds ratio 1.174, 95% confidence interval 1.061-1.298; P=0.002) and good collateral circulation was an independent protective factor for good outcome (odds ratio 0.095, 95% confidence interval 0.038-0.239; P<0.001). Conclusions:Good collateral circulation was more common in patients with perforator pattern, and poor collateral circulation was more common in patients with watershed pattern. Good collateral circulation was independently associated with the good clinical outcome in acute ischemic stroke patients with anterior circulation intracranial atherosclerosis.

Ginsenoside Ro decreased measures of inflammation, aging, oxidants and thrombus formation in a previous study. To measure ginsenoside Ro content in red ginseng from different years, an optimized extraction method was developed to determine ginsenoside Rg₁, Re, Rb₁ and Ro content by HPLC in 43 batches of red ginseng from different origins, growing years and manufacturers. The results indicate that the best extraction method was to ultrasonify a 1 g sample in 70% methanol for 50 min. The total running time of the optimized gradient was 50 min using a C18 core-shell column and was half the time described in the Chinese Pharmacopoeia, 2015 edition. The separation resolution of all of targeted compounds was greater than 1.6. The peak shape of ginsenoside Ro was optimal when the mobile phase consisted of acetonitrile and water with 0.1% phosphoric acid. The content of ginsenoside Ro was in the range of 0.11% to 0.43%, and the average content was 0.26%, which was higher than that of ginsenoside Rg₁ and Re. The ratio of ginsenoside Ro and Re as a threshold could be used to discriminate red ginseng from different growing years; in addition, 100%, 94.4% and 46.6% of red ginseng from six, five and four years exceeded the threshold of 1.3. Our optimized analytical method for characterization of red ginseng is convenient and shortens the assay time.

Classical famous prescription Dihuang Yinzi is widely used in modern clinical practice，and can treat many kinds of diseases，especially the diseases of nervous system in internal medicine. Its clinical effect is accurate，but it has not been converted into Chinese patent medicine preparations. Therefore，the authors have collected ancient traditional Chinese medicine（TCM） literatures of Dihuang Yinzi by the methods of bibliometrics，and selected and sorted out 254 pieces of effective data， involving 144 ancient books of TCM，and systematically summarized and analyzed the historical development origin，main treatment syndrome，formula making principle，dosage，preparation method，decoction method and medicine taking method of Dihuang Yinzi，in order to provide the ancient literary evidence support for the development and clinical application of classic famous prescriptions. It is found that Dihuang Yinzi was from Xuanming Lunfang written by LIU He-jian，a doctor of Jin dynasty. It was composed of 12 kinds of herbs，namely Rehmanniae Radix Praeparata，Morindae Officinalis Radix，Corni Fructus，Cistanches Herba，Dendrobii Caulis，Aconm Lateralis Radix Praeparaia，Schisandrae Chinensis Fructus，Cinnamomi Ramulus，Poria，maimendong，Acori Calami Rhizoma and Polygalae Radix, and mainly used for the treatment of Yinfei. The later records of Dihuang Yinzi mostly followed the prescription composition and main treatment set forth in Xuanming Lunfang，and its clinical application was expanded. In the 199 articles with the indications for disease treatment，Yinfei was the most commonest indication, and took up about half of the total，which was followed by stroke，taking up about two fifths of the total. It was also used for the treatment of sudden aphonia，flaccidity syndrome，vertigo，enuresis. Dihuang Yinzi has a wide range of treatment，but the pathogenes is always belongs to "the deficiency of water and fire in the kidney". The recipe of Dihuang Yinzi was unique，and can be used to treat both the upper and lower parts of the body，as well as both the outward symptoms and root causes of an illness at the same time， in particular，it mainly focuses on the treatment of the lower and the root. Among the 56 literatures with drug dosage records，about one third of them inherited the records of Xuanming Lunfang: "Equal division，the top is the end，3 qian for each dose." The dosage was generally light. The preparations are mostly decoction and boiled powder. In the decocting and taking methods，it was suggested that "turbid medicine shall be boiled for a short time，and taken after several boilings，with no limit to time."