Ischemic cerebrovascular disease is a common and frequently emergency, and leads to disability and death. In recent years,inflammatory reaction is thought as the key to secondary injury after cerebral ischemia/reperfusion. Inflammatory cells, as the target of inflammatoryfactors, infiltrate and gather in the ischemic areas leading to secondary neurological injury and enlarge the infarct size. Acupuncturemay adjust the activity of inflammatory cells.

A new fusion expression vector, pED-P8-Hi-lys was designed and constructed. It includes four parts, a 20 peptide sequence of hirudin that can maintain anticoagulant activity, the C-terminus of asparaginase as a fusion partner, basic octopeptide (KRKRKKSR) that makes the fusion partner easy to remove, and the unique acid-labile aspartyl-prolyl bond. It was transformed into E. coli BL-21 and the fusion protein (AnsB-C-P8-Hi-lys) was expressed effectively as inclusion bodies after inducing by lactose. The objective peptide Hi-lys was purified by means of cell disruption, washing, ethanol precipitation, acid hydrolysis, and DEAE-cellulose 52 column chromatography. The antithrombin activity of the purified Hi-lys peptide was about 50 ATU/mg by thrombin activity assays.

AIM　The purpose is to construct D-hydantoinase genetic engineering strain for the purpose of the industrial production of D-p-hydroxyphenylglycine. METHODS　D-hydantoinase gene was created from Pseudomonas putida 9801 by PCR technique and inserted into pMD18-T vector. The recombinant plasmid was transformed into several Escherichia coli strains. The positive transformants with D-hydantoinase activity were obtained by the two step screening, digoxigenin DNA labeling in situ hybridization and D-hydantoinase activity assay. RESULTS　The D-hydantoinase activity of the genetic engineering strain E.coli BL21/pMD-dht was 1700 U*L-1 and increased as high as 8 times compared with those of wild-type strain Pseudomonas putida 9801. The subunit molecular weight of recombinant D-hydantoinase was about 53 kDa measured by SDS-PAGE. The amount of the recombinant D-hydantoinase was about 20 percent of total bacterial soluble proteins. CONCLUSION　The genetic engineering strain E.coli BL21/pMD-dht possesses the initial industrial production prospects.

Objective To investigate the effects of 3％ and 7.5％ hypertonic saline (HS) on hemorrhagic shock patients in Emergency Department.Methods From December 2008 to February 2012,patients older than 15 years with severe trauma and systolic blood pressure (SBP) ≤70 mmHg or 70 to 90 mmHg with heart rate≥ 108 per minute were divided into three groups randomly (random number).Group A:patients treated with 3％ hypertonic saline (HS) 300 mL + lactated Ringer' s solutions (LRS).Group B:patients treated with 7.5％ HS 300 mL + LRS.Group C:patients treated with LRS.The mean arterial pressure (MAP),blood pressure (BP),heart rate (HR) were recorded before infusion and at 10,30,45,60 minutes successively after infusion.Incidence of complications and mortality rates were compared between groups.Results Atotal of 148 patients were enrolled in this study.Compared with LRS grouop,MAP was restored more promptly and maintained persistently in 3％ HS group and 7.5％ HS group,and the total volume of fluid infused was decreased to almost 50％ of LRS in the first 1 hour.No significant differences in MAP levels were observed between group A and B except 30 minutes after infusion.Single bolus of 7.5％ HS infusion resulted in increased of HR to mean 127 beats per minute at 10 minutesafter fluid resuscitation.Higher incidence of arrhythmia and transient hypotension occurred in 7.5％ HS group.There were no statistical differences of changes of electrolytic indices,mortality rates,incidences of ARDS and MODS among three groups.Conclusions Resuscitation with 3％ HS provide similar benefits and lower risk of complications compared with 7.5％ HS and LRS.This study demonstrates the practicability and safety of 3％ HS for fluid resuscitation of patients with hypovolemic shock.

Objective:To evaluate the role of extracellular signal-regulated kinase (ERK)1/2 in glutamate-induced ferroptosis in PC12 cells.Methods:PC12 cells were divided into 6 groups ( n=21 each) using a random number table method: control group (C group), glutamategroup (Glu group), glutamate+ ERK1/2 over-expression group (Glu+ ERK1/2-OE group), glutamate+ ERK1/2 plasmid empty vector group (Glu+ Vec group), glutamate+ ERK1/2 knockdown group (Glu+ si-ERK1/2 group)and glutamate+ ERK1/2 SiRNA negative control group (Glu+ si-NC group). Cells were treated with glutamate at a final concentration of 6 mmol/L for 72 h in Glu group and with the equal volume of PBS buffer for 72 h in C group. Glu+ ERK1/2-OE group was transfected with ERK1/2 overexpression plasmid, Glu+ Vec group was transfected with plasmid empty vector, and Glu+ si-ERK1/2 group was transfected with ERK1/2 siRNA, Glu+ si-NC group was transfected with siRNA negative control for 48 h, and then glutamate at a final concentration of 6 mmol/L was added and cells were treated for 72 h. The cell viability, lactic dehydrogenase (LDH)activity and contents of glutathione (GSH), ferrous ions and malondialdehyde (MDA) were measured by enzyme-linked immunosorbent assay. Mitochondrial membrane potential (MMP) and lipid reactive oxygen species (Lip-ROS) were measured by flow cytometry. Results:Compared with C group, the cell viability, GSH content and MMP were significantly decreased, and the LDH activity, ferrous ions content, MDA content and Lip-ROS levels were increased in Glu group ( P<0.05). Compared with Glu+ Vec group, the cell viability, GSH content and MMP were significantly increased, and the activity of LDH, contents of ferrous ions and MDA, and Lip-ROS levels were decreased in Glu+ ERK1/2-OE group( P<0.05). Compared with Glu+ si-NC group, the cell viability, GSH content and MMP were significantly decreased, and the LDH activity, contents of ferrous ions and MDA, and Lip-ROS level were increased in Glu+ si-ERK1/2 group ( P<0.05). Conclusions:ERK1/2 is involved in glutamate-induced ferroptosis in PC12 cells.

Objective:To retrospectively observe the effect of oxycodone on acute postoperative pain in patients undergoing thoracoscopic surgery.Methods:A retrospective analysis was conducted on the clinical data of 404 patients undergoing thoracoscopic thoracic surgery under combined general anesthesia at the Xiangya Hospital, Central South University from April 1, 2020 to September 30, 2021. They were divided into A group ( n=99, oxycodone group) and B group ( n=305, control group) based on whether oxycodone was used during the surgery. The two groups of patients were further matched 1∶1 using the nearest neighbor matching method. We compared the Visual Analogue Scale (VAS) of activity and resting pain and the incidence of moderate to severe pain between two groups of patients 24 hours after surgery, and observed the incidence and severity of pain related adverse reactions such as nausea, vomiting, itching, and dizziness. Resultsl:After matching the propensity scores of the two groups of patients, the balance was good ( SMD<0.20). There was no statistically significant difference between the groups in age, gender, body mass index, American Society of Anesthesiologist (ASA) grade, surgical time, intraoperative bleeding, and the use of antiemetics and analgesics during the perioperative period (all P>0.05). Compared with the control group, patients in the group A had a resting VAS [(2.03±1.61)points vs (1.62±1.31)points, P=0.049], and activity VAS [(4.13±1.72)points vs (3.51±1.79)points, P=0.013] was even lower, and the incidence of moderate to severe pain (VAS≥4) during activity was lower [59.6%(59/99) vs 37.4%(37/99), P=0.003]. There was no statistically significant difference in the incidence of analgesic related adverse reactions between the two groups ( P>0.05). Conclusions:Intravenous injection of oxycodone can effectively alleviate acute pain in patients undergoing thoracoscopic surgery within 24 hours, and reduce the incidence of moderate to severe pain during activity.

Objective:To study the short-term clinical outcomes of different courses of antenatal corticosteroids (ACS) for preterm twins.Methods:From January 2017 to December 2021, preterm twins with gestational age (GA) 24-34 weeks admitted to the neonatal ward of our hospital and received ACS were retrospectively studied. The infants were assigned into single-course group, partial-course group and multiple-course group according to ACS courses. The short-term clinical outcomes were compared among the groups. SPSS software version 25.0 was used for statistical analysis.Results:A total of 286 infants were enrolled in this study, including 128 in single-course group, 89 in partial-course group and 69 in multiple-course group. Compared with single-course group, the risks of neonatal respiratory distress syndrome (RDS) in both partial-course group ( OR=2.332, 95% CI 1.028-5.293, P=0.043) and multiple-course group ( OR=3.872, 95% CI 1.104-13.584, P=0.034) were higher. The birth length in multiple-course group ( β=-0.016, 95% CI -0.029 - -0.002, P=0.024) was lower than single-course group. Conclusions:The risks of neonatal RDS in preterm twins are higher in partial-course and multiple-course of ACS. A full course of ACS should be used to prevent neonatal RDS until further evidence of effectiveness is available.

Objective    To investigate the short-term therapeutic effect of neoadjuvant immunotherapy combined with chemotherapy in the locally advanced esophageal squamous cell carcinoma. Methods    The clinical data of patients with esophageal squamous cell carcinoma treated with neoadjuvant treatment in Gaozhou People's Hospital from August 2019 to October 2020 were retrospectively analyzed. According to the different treatments, the patients were divided into two groups: a neoadjuvant immunotherapy combined with chemotherapy group (NIC group) and a neoadjuvant chemoradiotherapy group (NC group). The baseline data, incidence of adverse events during treatment, perioperative indicators, postoperative pathological remission rate and incidence of postoperative complications were compared between the two groups. Results    Totally 33 patients were enrolled, including 15 males and 18 females, with an average age of 62.37±7.99 years. There were 17 patients in the NIC group and 16 patients in the NC group. In the NIC group, the carcinoma was mainly located in the middle and lower esophagus, with 5 paitents in stage Ⅱ, 9 patients in stage Ⅲ, and 3 patients in stage Ⅳa. In the NC group, the carcinoma was mainly located in the upper-middle esophagus, with 1 patient in stage Ⅱ and 15 patients in stage Ⅲ. During the neoadjuvant treatment, there was no significant difference in the occurrence of bone marrow suppression or gastrointestinal reactions between the two groups (P>0.05). There were 4 immune-related rashes in the NIC group and 1 esophageal perforation in the NC group. Fourteen (82.35%) patients in the NIC group and 12 (75.00%) patients in the NC group completed the operation on schedule. The postoperative ICU stay time and chest tube indwelling time in the NIC group were shorter than those in the NC group (P<0.05). There were 5 patients of complete remission in the NIC group, and 6 patients in the NC group. There was no significant difference in the pathological regression grade or residual tumor cells between the two groups (P>0.05). There was no significant difference in the incidence of anastomotic fistula, thoracic gastric fistula, bronchial mediastinal fistula, abdominal distension, pulmonary infection, stroke, or hoarseness during the perioperative period between the two groups of patients who completed the operation (P>0.05). In the NC group, 2 patients died during the perioperative period because of thoracic gastric fistula complicated by severe infection. Conclusion    Neoadjuvant immunotherapy combined with chemotherapy dose not significantly increase the occurrence of adverse events and shows a good rate of pathological remission, which indicates that the neoadjuvant immunotherapy combined with chemotherapy is a safe, feasible and potential new treatment model.