BACKGROUND:Curcumin is the main active ingredient of turmeric and has significant medicinal value in anti-tumor,anti-inflammatory,antioxidant and other aspects.However,its poor water solubility,unstable chemical properties and easy decomposition lead to difficulty in extracting curcumin and low extraction yield.Therefore,it is particularly important to optimize the curcumin extraction method.OBJECTIVE:To enhance the extraction yield and utilization value of curcumin and optimize the curcumin extraction process and curcumin nanoparticle preparation process.METHODS:Curcumin was extracted from turmeric by ethanol extraction,ultrasonic extraction,ionic liquid extraction,enzyme extraction,and ionic liquid combined with ultrasonic assisted enzyme extraction.The curcumin extraction yield was detected by high performance liquid chromatography;the best extraction method was determined,and subsequent process optimization experiments were carried out.The curcumin extraction yield was the response value with the type of ionic liquid,reaction temperature,ultrasonic time,liquid-to-solid ratio,ionic liquid concentration,and enzyme-drug mass ratio as parameters.The optimal production process of ionic liquid combined with ultrasonic assisted enzyme extraction was determined by single factor combined response surface experiment.The optimal process for preparing curcumin nanoparticles by ionic crosslinking method was determined by single factor combined response surface experiment with acetic acid concentration,chitosan to sodium tripolyphosphate mass ratio,stirring rate,curcumin mass concentration,sodium tripolyphosphate mass concentration,and chitosan mass concentration as parameters,and drug encapsulation efficiency as response value.Curcumin nanoparticles were prepared under the optimal process,and the particle size,polydispersity index,Zata potential value,drug loading,stability,hemolysis rate,and antioxidant capacity in vivo and in vitro of the nanoparticles were detected.RESULTS AND CONCLUSION:(1)Among the five extraction methods,the curcumin yield of ionic liquid combined with ultrasound-assisted enzyme extraction was the highest,and this method was selected as the curcumin extraction method for subsequent experiments.The results of single factor combined response surface experiment showed that the optimal process for curcumin extraction was:ionic liquid selected 1-hexyl-3-methylimidazolium chloride,reaction temperature 55 ℃,liquid-to-solid ratio 40 mL/g,ultrasound time 57 minutes,ionic liquid concentration 57％,enzyme-drug mass ratio 3.5:10,and the obtained turmeric extraction yield was 3.10％.The optimal preparation process of curcumin nanoparticles was:glacial acetic acid concentration 0.5％,chitosan and sodium tripolyphosphate mass ratio 5.0:1,stirring speed 150 r/min,curcumin mass concentration 2.23 mg/mL,sodium tripolyphosphate mass concentration 1.45 mg/mL,chitosan mass concentration 3.63 mg/mL,and the obtained drug encapsulation efficiency was 90.61％.(2)The drug loading of curcumin nanoparticles was(14.49±0.23)％,the average particle size was(76.95±1.65)nm,the polydispersity coefficient was 0.15±0.02,and the Zata potential value was(32.37±1.46)mV.The curcumin nanoparticles had good stability and blood compatibility,did not induce hemolysis,and had stronger antioxidant capacity in vivo and in vitro than free curcumin.(3)The results show that the process optimization not only solves the problems of low extraction yield,poor solubility,and low bioavailability of curcumin,but also enhances its antioxidant activity in vivo and in vitro.

BACKGROUND:Curcumin is the main active ingredient of turmeric and has significant medicinal value in anti-tumor,anti-inflammatory,antioxidant and other aspects.However,its poor water solubility,unstable chemical properties and easy decomposition lead to difficulty in extracting curcumin and low extraction yield.Therefore,it is particularly important to optimize the curcumin extraction method.OBJECTIVE:To enhance the extraction yield and utilization value of curcumin and optimize the curcumin extraction process and curcumin nanoparticle preparation process.METHODS:Curcumin was extracted from turmeric by ethanol extraction,ultrasonic extraction,ionic liquid extraction,enzyme extraction,and ionic liquid combined with ultrasonic assisted enzyme extraction.The curcumin extraction yield was detected by high performance liquid chromatography;the best extraction method was determined,and subsequent process optimization experiments were carried out.The curcumin extraction yield was the response value with the type of ionic liquid,reaction temperature,ultrasonic time,liquid-to-solid ratio,ionic liquid concentration,and enzyme-drug mass ratio as parameters.The optimal production process of ionic liquid combined with ultrasonic assisted enzyme extraction was determined by single factor combined response surface experiment.The optimal process for preparing curcumin nanoparticles by ionic crosslinking method was determined by single factor combined response surface experiment with acetic acid concentration,chitosan to sodium tripolyphosphate mass ratio,stirring rate,curcumin mass concentration,sodium tripolyphosphate mass concentration,and chitosan mass concentration as parameters,and drug encapsulation efficiency as response value.Curcumin nanoparticles were prepared under the optimal process,and the particle size,polydispersity index,Zata potential value,drug loading,stability,hemolysis rate,and antioxidant capacity in vivo and in vitro of the nanoparticles were detected.RESULTS AND CONCLUSION:(1)Among the five extraction methods,the curcumin yield of ionic liquid combined with ultrasound-assisted enzyme extraction was the highest,and this method was selected as the curcumin extraction method for subsequent experiments.The results of single factor combined response surface experiment showed that the optimal process for curcumin extraction was:ionic liquid selected 1-hexyl-3-methylimidazolium chloride,reaction temperature 55 ℃,liquid-to-solid ratio 40 mL/g,ultrasound time 57 minutes,ionic liquid concentration 57％,enzyme-drug mass ratio 3.5:10,and the obtained turmeric extraction yield was 3.10％.The optimal preparation process of curcumin nanoparticles was:glacial acetic acid concentration 0.5％,chitosan and sodium tripolyphosphate mass ratio 5.0:1,stirring speed 150 r/min,curcumin mass concentration 2.23 mg/mL,sodium tripolyphosphate mass concentration 1.45 mg/mL,chitosan mass concentration 3.63 mg/mL,and the obtained drug encapsulation efficiency was 90.61％.(2)The drug loading of curcumin nanoparticles was(14.49±0.23)％,the average particle size was(76.95±1.65)nm,the polydispersity coefficient was 0.15±0.02,and the Zata potential value was(32.37±1.46)mV.The curcumin nanoparticles had good stability and blood compatibility,did not induce hemolysis,and had stronger antioxidant capacity in vivo and in vitro than free curcumin.(3)The results show that the process optimization not only solves the problems of low extraction yield,poor solubility,and low bioavailability of curcumin,but also enhances its antioxidant activity in vivo and in vitro.

Objective:To gain an in-depth understanding of the psychological safety experiences and changes of new nurses in Neurosurgery Department, providing insights for improving their psychological safety.Methods:A purposive sampling method was used to select new nurses from the Neurosurgery Department at Zhejiang Enze Medical Center Enze Hospital in July to August 2023. Semi-structured interviews were conducted, and Colaizzi's 7-step analysis method was applied to organize and analyze the interview data, summarizing and refining the themes.Results:A total of eleven new nurses from the Neurosurgery Department were included in the study. A total of five themes were identified: insufficient awareness of psychological safety, self-doubt, lack of interpersonal safety, fear and worry, and coping and adaptation.Conclusions:New nurses in Neurosurgery Department often experience a lack of psychological safety during their development. It is crucial to focus on their stage-specific characteristics, enhance new nurse training, create a safe environment, assist them in adopting correct coping strategies, and improve their mental health.

Objective:To gain an in-depth understanding of the psychological safety experiences and changes of new nurses in Neurosurgery Department, providing insights for improving their psychological safety.Methods:A purposive sampling method was used to select new nurses from the Neurosurgery Department at Zhejiang Enze Medical Center Enze Hospital in July to August 2023. Semi-structured interviews were conducted, and Colaizzi's 7-step analysis method was applied to organize and analyze the interview data, summarizing and refining the themes.Results:A total of eleven new nurses from the Neurosurgery Department were included in the study. A total of five themes were identified: insufficient awareness of psychological safety, self-doubt, lack of interpersonal safety, fear and worry, and coping and adaptation.Conclusions:New nurses in Neurosurgery Department often experience a lack of psychological safety during their development. It is crucial to focus on their stage-specific characteristics, enhance new nurse training, create a safe environment, assist them in adopting correct coping strategies, and improve their mental health.

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

Objective To investigate the distribution characteristics of HIV-1 subtypes,the status of transmitted drug resistance(TDR),and the influencing factors of TDR in treatment-naive patients with AIDS who are hospitalized.Methods Treatment-naive patients with AIDS who were admitted to the Infectious Disease Department,Public Health Clinical Center of Chengdu between January 2020 and December 2022 were enrolled in the study.The diagnosis and confirmation diagnosis of all the subjects were made at the same hospital.Blood samples were collected from the subjects before antiretroviral therapy(ART).The in-house method was used for HIV gene amplification and sequencing.A phylogenetic tree was constructed to analyze the HIV-1 subtypes.The Stanford HIV Drug Resistance Database was used to conduct an online comparative analysis of the drug resistance mutation sites and to determine the types and levels of drug resistance.The distribution characteristics of HIV-1 subtypes,the occurrence of TDR,and the influencing factors of TDR were analyzed.Results A total of 213 patients were included in the study and their blood samples were collected.HIV-1 subtypes were successfully amplified in 83.10％(177/213)of the subjects.Ten HIV subtypes were identified,with CRF07_BC being the most common subtypes,accounting for 43.50％(77/177),which was followed by CRF01_AE at 37.85％.Unique recombinant forms(URFs)were relatively uncommon,accounting for 8.47％.The other subtypes accounted for 10.17％.These 4 categories of HIV-1 subtypes were distributed with statistically significant differences in different age groups(P=0.024).Further analysis revealed significant differences in the distribution of the HIV-1 subtypes of CRF01_AE and URFs between the groups of patients aged 30-50 years and those over 50.In addition,URFs accounted for a higher proportion in patients aged 30 to 50 years(P=0.008).The incidences of TDR were 6.49％,8.96％,13.33％,and 5.56％for CRF07_BC,CRF01_AE,URFs,and other subtypes,respectively,showing no significant difference(P＞0.05).The overall TDR was 6.57％.The TDR for non-nucleoside reverse transcriptase inhibitors(NNRTIs)was 5.16％,and the main mutation sites were V179D/E,E138A/G,V106M/I,and Y181C.The TDR for nucleoside reverse transcriptase inhibitors(NRTIs)was 1.88％,and the main mutation site was M184V.One patient was found to be resistant to both NNRTIs and NRTIs.The highly resistant rate was 4.23％,moderate resistance was 0.47％,and low resistance was 1.88％.No significant effects of the specific years,demographic characteristics,transmission route,baseline condition,and opportunistic infections on TDR were found in this study(P＞0.05).Conclusions The HIV-1 subtypes are diverse and complex in treatment-naive patients with AIDS who were hospitalized.The overall prevalence of TDR is relatively high.It is necessary to strengthen HIV drug resistance testing to optimize ART treatment and reduce the risk of drug resistance transmission.

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26％,and the intra-and inter-batch co-efficients of variation were both less than 10％.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.

Objective:To construct a predictive model of radiomics based on laparoscopic grayscale ultrasound features and investigate its value in predicting microvascular invasion (MVI) of hepatocellular carcinoma (HCC) during laparoscopic liver resection.Methods:A total of 74 patients (74 lesions)with HCC confirmed by postoperative pathology, who underwent a laparoscopic ultrasonography during laparoscopic hepatectomy were prospectively enrolled in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from March 2022 to August 2023. The general clinical information of the patients was recorded, and the features were extracted and screened from tumor regions in gray-scale ultrasound images, and eventually the radiomics prediction models were constructed, respectively. Pathological results were used as gold standard to compare the effectiveness of different models in predicting MVI.Results:In the 74 HCC lesions, 12 lesions were MVI positive.The MVI imaging prediction model of HCC lesions was constructed from the screened clinical features, laparoscopic gray scale ultrasound image features, as well as combined screened clinical features, respectively. The obtained data sets were randomly divided into 5 parts (4 parts with 15 lesions, 1 part with 14 lesions), and the effectiveness of the model was trained and tested by the method of 5 folds interaction validation. The performance of support vector machine(SVM) radiomics model based on the characteristics of laparoscopic gray scale ultrasound in predicting the MVI of HCC was the best. Compared with clinical model and combined Adaboost model, the SVM, radiomics model had higher area under ROC curve (0.836 vs 0.696, 0.804), accuracy (0.852 vs 0.687, 0.838), sensitivity (0.900 vs 0.900, 0.833) and specificity (0.837 vs 0.644, 0.838). Conclusions:The radiomics model based on the characteristics of laparoscopic gray-scale ultrasound is an innovative potential approach to predict the MVI status of HCC lesions during laparoscopic hepatectomy.

The clinical manifestations of tuberculous meningoencephalitis(TBME)are varied,and diagnostic tests for Mycobacterium tuberculosis show limited sensitivity and specificity,often failing to provide timely diagnostic support,which can delay prompt treatment.Reports on anti-neurexin-3α antibody-mediated autoimmune encephalitis remain scarce,and there have been no documented cases of TBME overlapping with anti-neurexin-3α antibody-associated encephalitis either domestically or internationally.This article presents a case treated at our hospital to contribute insights for improving the diagnosis and treatment of such conditions in the future.The patient,a middle-aged woman,presented with a one-month history of headache and incoherent speech,along with one day of left-sided limb weakness.Lumbar puncture results revealed significantly elevated cerebrospinal fluid(CSF)pressure,an increased lymphocyte count,and reduced glucose and chloride levels.Enhanced MRI showed multiple intracranial lesions,basal meningeal enhancement,and marked hydrocephalus,supporting an initial diagnosis of TBME.Concurrently,both blood and CSF tests were positive for anti-neurexin-3α antibodies.After comprehensive treatment,including anti-tuberculosis therapy,high-dose immunoglobulin,and corticosteroids,the patient's clinical symptoms,CSF findings,and MRI results improved,and she was discharged.