Objective To construct p66shc gene interfering lentivirus vectors recombination and transfect it to 293T cells ,RNA interfering was carried out to induce p66shc gene silence ,so as to provide basis for further study of the p66shc function .Methods Screening of three RNA targets which were named after p66shc‐shc1 ,p66shc‐shc2 ,p66shc‐shc3 ,cloned into the pLenR‐GPH vec‐tor ,which contained green fluorescent protein(GFP) and transformed into DH5αcells .The positive clone were picked out for right sequencing and transfected to 293T cells with pRsv‐REV ,pMDlg‐pRRE ,pMD2G .The expression of GFP in inverted fluorescence microscope confirmed the virus packaging success .Fluorescence quantitative PCR and Western blot technology were used to investi‐gate the expression of p66shc at the molecular and protein levels ,p66shc‐shc1 target of effective silencing p66shc gene was selected to prepare for subsequent tests .Results The shRNA lentivirus vector was constructed which could express p66shc and was trans‐fected into 293T cells successfully .Fluorescence quantitative PCR and Western blot technology were used to investigate p66shc gene silence by RNA interference .Conclusion The lentivirus RNAi vector of targeted expression p66shc could induce p66shc gene si‐lence at the molecular and protein levels after transfected into 293T cells by RNA interference .

Objective To study the protective effect of sodium pyruvate solution to intestinal barrier in Hemorrhagic shock rats by intraperitoneal resuscitation.Methods A total of 40 Male SD ratswere healthy and clean,were randomly (random number) divided into four groups:sham operation group (group Ⅰ),hemorrhagic shock model group (group Ⅱ),the recovery of the compound sodium chloride intravenous + intraperitoneal recovery of the PD-2 liquid group (group iiⅢ),the recovery of the compound sodium chloride intravenous + sodium pyruvate abdominal recovery group (group Ⅳ) (n =10).In addition to the group Ⅰ,the other three groups are used the HS model by The Wiggers improvement prepared,group Ⅲ,Ⅳ were used the same amount of intravenous recovery,At the same time to the abdominal cavity were respectively injected 20 mL 2.5％ PD-2 liquid and 20 m 2.5％ sodium pymvate solution.After 2 h,each group were taken arterial blood measured arterial blood gases (pH,PaCO2,PaO2) and D-lactic acid,small intestine specimens under the light microscope to observe the morphological changes of the rat small intestinal mucosa.Results Blood gas analysis:group Ⅲ and Ⅳ was significantly increased compared with group Ⅱ in pH levels and PaO2 content (P ＜ 0.01),and comoared to group Ⅲ,pH levels and PO2 levels of group Ⅳ increased more significantly (P ＜ 0.05) ; D-LA content:Group Ⅲ,Ⅳgroup is significantly lower than group Ⅱ (P ＜ 0.01) ; compared with group Ⅲ and group Ⅳ,D-LA significantly decreased (P ＜ 0.05).Light microscope,the group Ⅱ,the rat small intestine layer of a high degree of villous edema,submucosal vascular collapse within the lamina propria glands focal area of necrosis,and large areas of mucosal villi necrosis,defects; group Ⅲ and group Ⅳof small intestinal damage compared to group Ⅱ significantly reduced; Ⅳgroup of the rat small intestine villi edema,lodging and the degree of necrosis in group Ⅲ.Intestinal epithelial damage index score:the Ⅳ group than in group Ⅱ and Ⅲ group (P ＜ 0.01 or P ＜ 0.05).Conclusions The venous recovery and intraperitoneal resuscitation can significantly reduce the plasma D-LA content of hemorrhagic shock in rats,correct acidosis,increase the oxygen partial pressure ; and intestinal mucosal barrier function,the protective of sodium pyruvate solution is better than the PD-2 liquid.

Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.
Chromatography, Reverse-Phase ; methods ; Glucosides ; analysis ; Ionic Liquids ; chemistry ; Monoterpenes ; analysis ; Oleanolic Acid ; analysis ; Paeonia ; chemistry ; Ultrasonics

Four kinds of ionic liquids were adopted to analyze the content of rubimaillin and alizarin in Rubia cordifolia roots with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 microm), was used. Methanol and 0.4% acetic acid-water as mobile phase with flow rate at 0.85 mL min(-1), gradient elution, detection wavelength at 250 nm, chromatographic column temperature was controlled at room temperature. The result showed that rubimaillin and alizarin had the highest extraction yield when the [ HMIM] PF6methanol solution concentration of 0.6 mol x L(-1) as extraction solvent and the conditions were solid-liquid ratio of 1:80 (g x mL(-1)). Under the optimal extraction conditions, the content of alizarin from 0.01 to 0.04 microg showed a good linearity (r = 0.9999), the average recovery was 97.12%, the content of rubimaillin from 0.41 to 1.35 microg showed a good linearity (r = 0.9999), the average recovery was 98.10%. This experiment adopted environmentally friendly reagent as extraction solvent, the extraction efficiency was improved, and the environmental pollution caused by organic solvent was avoided, the harm of human body aslo was reduced. This method was simple and reliable, its repeatability was also very good, which had an important significance in the study of traditional Chinese medicine active ingredient extraction methods.
Anthraquinones ; analysis ; Chromatography, Reverse-Phase ; methods ; Ionic Liquids ; chemistry ; Pyrans ; analysis ; Rubia ; chemistry ; Ultrasonics

Objective To investigate the protective effects of intra-peritoneal fluid resuscitation on small intestinal mucosa in rats with hemorrhagic shock. Method Fifty Sprague-Dawley (SD) rats were randomly (random number) divided into five groups, namely sham operation group (group I ), hemorrhagic shock group (group Ⅱ ), intra-venous fluid resuscitation group (group Ⅲ ) . intravenous fluid resuscitation plus intra-peritoneal saline resuscitation (group Ⅳ ) and intravenous fluid resuscitation plus intra-peritoneal PD-2 solution resuscitation group (group Ⅴ ). The rats of 5 groups were processed with cannulations of right common carotid artery, right femoral vein and left femoral artery with systemic heparinization. The rat models of hemorrhagic shock were established with modified Wigger' s method by which the blood exsanguinated from left femoral artery. The rats of group Ⅲ were resuscitated with shed blood plus twice equal volume of Ringer's solution after modeling of hemorrhagic shock.The rats of group Ⅳ and group Ⅴ were administered intra-peritoneally with 30 mL saline and 30 mL of 2.5% PD-2 solution, respectively as adjuncts to those used in the group Ⅲ . The specimens of blood and small intestine of rats of all groups were collected 60-120 minutes after modeling and resuscitation. The activity of plasma diamine oxidase (DAO) was determined with chromatometry, the level of plasma D-lactic acid (D-LA) with spectorophotometry and the level of plasma lipopolysaccharide (LPS) with nephelometry. The histopathological and ultrastructure changes of small intestine tissue of rats were observed under light microscope and electronic microscope. Results There were remarkable differences in activity of DAO, and the levels of D-LA and IPS in rats between those ingroup Ⅱ and group I (P <0.01), and between those in group V and groups Ⅱ , Ⅲ or Ⅳ (P <0.05 or P < 0.01) The pathomorphology and ultra-structure of small intestine tissues were severely damaged in group Ⅱ compared with those in group Ⅰ , and those markedly lessened in group V compared with groups Ⅱ , Ⅲ and Ⅳ . Conclusions Intraperitoneal fluid resuscitation with PD-2 solution can significantly protect the integrity of intestinal mucosa and the normal permeability of intestinal wall, and blunts the histopathological changes, and restrains bacterial translocation from gut and reduces the level of plasma endotoxin.

OBJECTIVETo investigate the current smoking regulations and their impacts on the environmental tobacco smoke (ETS) levels inside restaurants and bars in Beijing.

METHODSTelephone survey was used to investigate the smoking regulations. TSI Sidepak AM510 was used to measure the level of fine particles less than 2.5 microns in diameter (PM2.5) in restaurants and bars. Analysis of variance and non-parametric rank tests were used to examine the association between indoor and outdoor PM2.5 levels and (1) smoking regulations; and (2) types of restaurants and bars.

RESULTSOf the 305 restaurants and bars surveyed, 27.9% had complete or partial smoking prohibiting rules. The average indoor PM2.5, level of the 92 restaurants and bars was 253.08 microg/m3 , 102.37% higher than the outdoor level. The average indoor and outdoor PM2.5 levels in the restaurants and bars with smoking ban regulations were 93.10 microg/m3 and 110.33 microg/m3 whole 289.34 microg/m3 and 128.40 microg/m3 in those without, respectively. The average indoor and outdoor PM2.5 levels of bars were 413.46 microg/m3 and 190.62 microg/m3, respectively, while in the western fast-food restaurants, they were 83.86 microg/m3 and 104.77 microg/m3, respectively. The outdoor PM2.5 levels were higher than the indoor levels in different classes of restaurants and bars. Furthermore, there was a significant positive correlation between PM2.5 levels and the number of smokers per cube meters (r = 0.47, P < 0.001).

CONCLUSIONSmoking regulations could effectively reduce the ETS level in restaurants and bars.

Air Pollution, Indoor ; analysis ; legislation & jurisprudence ; China ; Environmental Monitoring ; Particulate Matter ; analysis ; Restaurants ; Smoking ; legislation & jurisprudence ; Tobacco Smoke Pollution ; analysis ; legislation & jurisprudence

Objective To report a case of synovial sarcoma of the liver and review the literature for improving the understanding of the disease.Methods The clinical data of a patient with liver synovial sarcoma admitted to the First Affiliated Hospital of Henan University were analyzed retrospectively.The imaging,pathological features,treatment and prognosis of this disease were summarized by searching the database(CNKI,Wanfang Data,PubMed,untill July 2022)and the literature results analyzed comprehensively.Results The patient was a 71-year-old female who was admitted to the hospital due to abdominal pain.Computed tomography(CT)scan showed a mass with mixed density in the right lobe and caudate lobe of the liver.The large cross section size was about 115 mm×87 mm and the mass showed continuous heterogeneous enhancement,being considered as malignant hepatic tumors with multiple metastasis of the liver and lung.Ultrasound-guided needle biopsy was performed,and microscopy showed the tumor cells were obvious atypical,and some were spindle-shaped.Immunohistochemistry showed that the patient was positive for vimentin(VIM),epithelial membrane antigen(EMA),methylation of histone at lysine 27(H3K27Me3),and negative for pan cytokeratin(CK-pan)and S-100,and pathological diagnosis of synovial sarcoma was made.The patient did not undergo subsequent treatment and was lost to follow-up after discharge.A total of 12 cases of hepatic synovial sarcoma were reported after searching the database.The clinical manifestations were abdominal pain or distention.The lesions were mostly located in the right lobe of the liver,usually large,heterogeneous density,and heterogeneous enhancement on enhanced CT or magnetic resonance imaging(MRI).Spindle-shaped cells were found at histopathologic examination.Immunohistochemistry showed the patient was positive for VIM,EMA,H3K27Me,B-cell leukemia/lymphoma-2(BCL-2)and transducer-like enhancer of split 1(TLE1).SS18-SSX fusion gene or SS18 gene isolation were detected.Eleven patients received surgical treatment,5 received adjuvant chemotherapy,and 4 had recurrence or metastasis during the follow-up period.Conclusions Synovial sarcoma of the liver is a rare malignant tumor of the liver.The clinical and imaging features are not specific.The diagnosis depends on pathology.At present,the main treatment is surgery,and comprehensive treatment such as adjuvant chemotherapy can be performed.The prognosis of the patient is poor.

To analyze the positivity rate and titer of antinuclear antibody (ANA), as well as nuclear pattern and target antigen of ANA in healthy pregnant women during early pregnancy in Qingdao area. A prospective cohort study design was used to include a total of 9 528 healthy pregnant women registered at the Women and Children′s Hospital Affiliated to Qingdao University from March 2023 to June 2024.Indirect immunofluorescence assay (IIF) was used to detect ANA, its titer and cell staining pattern. Fifteen specific antibodies were tested using the magnetic bar code immunofluorescent luminescence method. Logistic regression model was used to analyze the risk factors of pregnancy with autoimmune disease(AID). The results showed that among 9 528 pregnant women in early pregnancy, 1 346 cases (14.1%) were positive of ANA, including 1 011 cases with a titer of 1∶100 (10.6%), 236 cases (2.5%) with a titer of 1∶320, and 99 cases (1.0%) were detected at a titer >1∶320. Among the 1 346 ANA-positive pregnant women, nuclear granular type accounted for the highest proportion (483 cases, 35.9%), followed by speckled type (347 cases, 25.8%) and cytoplasmic type (176 cases, 13.1%).Then, pregnant women with ANA titers ≥1∶100 were detected 15 specific antibodies.Anti-SSA was tested in 121 cases accounted for the majority, followed by 110 cases with anti-Ro-52, 56 cases with anti-SSB, 51 cases with anti-mitochondrial M2 subtype antibodies and 37 cases with anti-centromere B. In conclusion,in healthy pregnant women in Qingdao area, ANA positivity rate was 14.1%, and the titer of ANA was mainly at 1∶100.The predominant nuclear patterns were nuclear granular and speckled types.The specific autoantibodies were mainly anti-SSA antibodies and anti-Ro-52 antibodies.The detection of ANA and specific autoantibodies is of great significance for early prediction, diagnosis, and intervention of autoimmune diseases during pregnancy.

To analyze the positivity rate and titer of antinuclear antibody (ANA), as well as nuclear pattern and target antigen of ANA in healthy pregnant women during early pregnancy in Qingdao area. A prospective cohort study design was used to include a total of 9 528 healthy pregnant women registered at the Women and Children′s Hospital Affiliated to Qingdao University from March 2023 to June 2024.Indirect immunofluorescence assay (IIF) was used to detect ANA, its titer and cell staining pattern. Fifteen specific antibodies were tested using the magnetic bar code immunofluorescent luminescence method. Logistic regression model was used to analyze the risk factors of pregnancy with autoimmune disease(AID). The results showed that among 9 528 pregnant women in early pregnancy, 1 346 cases (14.1%) were positive of ANA, including 1 011 cases with a titer of 1∶100 (10.6%), 236 cases (2.5%) with a titer of 1∶320, and 99 cases (1.0%) were detected at a titer >1∶320. Among the 1 346 ANA-positive pregnant women, nuclear granular type accounted for the highest proportion (483 cases, 35.9%), followed by speckled type (347 cases, 25.8%) and cytoplasmic type (176 cases, 13.1%).Then, pregnant women with ANA titers ≥1∶100 were detected 15 specific antibodies.Anti-SSA was tested in 121 cases accounted for the majority, followed by 110 cases with anti-Ro-52, 56 cases with anti-SSB, 51 cases with anti-mitochondrial M2 subtype antibodies and 37 cases with anti-centromere B. In conclusion,in healthy pregnant women in Qingdao area, ANA positivity rate was 14.1%, and the titer of ANA was mainly at 1∶100.The predominant nuclear patterns were nuclear granular and speckled types.The specific autoantibodies were mainly anti-SSA antibodies and anti-Ro-52 antibodies.The detection of ANA and specific autoantibodies is of great significance for early prediction, diagnosis, and intervention of autoimmune diseases during pregnancy.

Objective:To examine the early to mid-term clinical outcomes of aortic valve repair in patients with bicuspid aortic insufficiency.Methods:This is a retrospective cohort study. The clinical data of 124 patients with BAV insufficiency who underwent aortic valve repair from January 2017 to June 2023 in the Department of Cardiovascular Surgery at Fuwai Hospital were analyzed retrospectively. There were 117 males and 7 females with an age of (38.1±12.7) years (range: 14 to 65 years). Depending on whether the aortic sinus was replaced or not, surgical approaches were divided into valve sparing root replacement (reimplantation, remodeling, modified remodeling) and isolated aortic valve repair (annuloplasty, isolated aortic valve leaflet repair). Perioperative and follow-up data were collected. Kaplan-Meier method was used to plot the curves of survival rate, free recurrence rate of massive aortic valve insufficiency and free re-operation rate, and Log-rank test was used for comparison between groups.Results:Among the surgeries, there were 47 cases of reimplantation, 8 cases of remodeling, 8 cases of modified remodeling, 48 cases of aortic annuloplasty (external annuloplasty in 22 cases, CV-0 annuloplasty in 26 cases), and 13 cases of isolated leaflet repair. Leaflet plication was the most used leaflet repair technique, used in 103 patients. The cardiopulmonary bypass time was (133.7±56.9) minutes (range: 48 to 461 minutes), and aortic cross-clamp time was (103.8±47.8) minutes (range: 25 to 306 minutes), with no surgical mortality. All patients underwent outpatient or telephone follow-up. The cumulative follow-up time was 340.3 person-years and the mean follow-up time was ( M (IQR)) 34.0 (25.5) months (range: 3 to 76 months). The 5-year survival rate was 98.4%, the 5-year freedom from significant insufficiency rate was 93.4% and the 5-year freedom from aortic valve reoperation rate was 95.6%. The subgroup analysis revealed a significantly better freedom from the significant insufficiency rate in the aortic valve annular reduction group compared to the non-reduction group ( P<0.01). Conclusions:Aortic valve repair in patients with bicuspid aortic insufficiency could obtain steady early to mid-term outcomes. Aortic annuloplasty can reduce the risk of recurrent aortic valve insufficiency in patients undergoing aortic repair.