[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

In this paper， by consulting the ancient and modern literature， the name， origin， quality evaluation， harvesting and processing， and others of the original animal and medicinal materials of Moschus were systematically sorted out and verified， in order to provide the basis for the development and utilization of the famous classical formulas containing Moschus. According to the textual research， musk deer was first recorded in Shanhaijing. Shennong Bencaojing was recorded as Moschus and all generations were used as the correct name， but there were also aliases such as Shefu， Xiangzhang and Xiangqizi. In ancient times， Moschus berezovskii， M. sifanicus and M. moschiferus were the main sources of Moschus， and the quality of Moschus produced in northwest China was better than that produced in the Yangtze River basin. In modern times， Moschus of M. moschiferus produced in northeast China， M. sifanicus produced in Gansu， Sichuan and other places， and M. berezovskii produced in Ningxia， Shaanxi and other places are regarded as genuine. In ancient times， gunshots， lassoes， arrow shots and other methods were generally used to hunt live musk deer， and the sachets were immediately cut off. Those with high quality were called Xiangshanhuo， and dried in the shade after harvesting， which was known as Maoke Shexiang. Cut open the sachet， remove the shell and dry preservation， commonly known as Moschus kernel. In modern times， the method of taking Moschus from the living body of cultured musk deer is adopted， that is， Moschus kernel is directly taken from its sachet， dried in the shade or dried in a closed dryer. This method realizes the sustainable utilization of Chinese herbal medicine resources， but attention should be paid to the frequency and quality of Moschus. The harvesting time is mostly after the autumnal equinox every year， and before the next summer， it is better to gather sachet in winter. In recent times， it is believed that the shell Moschus is dry， full， thin， elastic， loose inside， many particles， strong and persistent aroma for the best， while the Moschus kernel is particle purple-black， powder yellow-brown， soft and oily texture， strong and persistent aroma for the best. The ancient processing method of Moschus was extracting kernels from the shell. After removing impurities， it is ground and used as medicine. Because its composition is not suitable for heating， the processing method is most common in preparations such as grinding into powder and putting into pills or powders， which has the effect of opening up the orifices and refreshing the mind， and it has continued to this day. Based on the research conclusions， it is suggested that the development of famous classical formulas containing Moschus， M. sifanicus， M. moschiferus and M. berezovskii should be used as the origins. According to the processing requirements specified in the original formula， it should be processed and used as medicine， while those without processing requirements should be used as raw products.

Objective: To construct a risk prediction model for poly victimization (PV) among rural left behind middle and high school students in Chaoshan, and to evaluate the prediction effect of the model, so as to provide scientific basis for early identification and prevention of PV among students. Methods: A questionnaire survey was conducted among 1 005 left behind students, selected from 7 middle and high schools in rural areas of Shantou City and Jieyang City by a stratified random cluster sampling method from January 2020 to September 2021, for the personal, family, external environmental factors, psychological factors (mental resilience, coping approaches, self esteem and social support) and PV situations. R software and Logistic regression were used to screen predictor variables to build a risk prediction model, and the area under the ROC curve (area under the curve, AUC), accuracy, precision, recall, F1 value and calibration curve were used to evaluate the model s effect. Results: The incidence rate of PV among left behind middle and high school students was 23.38%. The results of Logistic regression analysis showed that physical illness or disability ( β =1.02), grade retention during the past year ( β =1.31), having no close partner ( β =1.00), self harm intention (seldom: β = 0.58 , occasionally: β =0.79), negative peer behavior ( β =0.90), family member smoking ( β =0.59), criminal offenses of parents ( β =1.04), witnessing school bullying ( β =0.78), house moving ( β =0.58), using venting ( β =0.34) and the coping style of patience ( β =0.28) were positively correlated with PV among left behind children in Chaoshan area, and family support in psychological flexibility ( β =-0.31) was negatively correlated with PV ( P <0.05). A nomogram prediction model was constructed for the meaningful variables included in the multivariate analysis, and the prediction model AUC was 0.88, the accuracy was 82.00 %, the precision was 77.78%, and the F1 value was 43.75%. The calibration plot fitted well, and the model had good discrimination and calibration. Conclusion The risk prediction model for left behind middle and high school students with PV has good predictive performance and is helpful for schools and communities to early identify high risk middle and high school students with PV.

Objective To study the effect of formononetin on the cell damage of glucose/oxygen deprivation/reoxy-genation glyconeurons via the PARP1 signaling pathway,and to offer theoretical support for the use of Caragana isoflavones in the treatment of cerebral ischemia-reperfusion injury.Methods In mouse neurons(HT22),a model of Oxygen-glucose deprivation/reoxygenation(OGD/R)was created.Western blot was used to detect the expres-sion of PARP1 and PARG in HT22 neurons at various time points of glucose-oxygen deprivation/reoxygenation,and the optimal time point of pathway modification was chosen.After OGD/R,HT22 cells were treated with form-ononetin,PARP1 inhibitor(PJ34),and PARG inhibitor,and six groups were developed:control group,control group+formononetin group,OGD/R group,OGD/R+formononetin group,OGD/R+PJ34 group,OGD/R+PARG inhibitor group.HT22 cells were grown normally without OGD/R therapy in the control group.The expres-sion levels of apoptotic factors and associated proteins in each group were determined using immunofluorescence and Western blot.Results PARP1 pathway was activated most obviously in HT22 cells after 3 hours of glucose and ox-ygen deprivation/reoxygenation.Under the condition of OGD/R 3 h,treatment with formononetin,PJ34 or PARG inhibitor could increase E3 ubiquitin ligase(Iduna),inhibit the expression of PARP1 and PARG pathway proteins,reduce the expression of AIF and P53,and increase the phosphorylation level of AKT protein.Conclusion Form-ononetin can block the PARP1/AIF/Akt signaling pathway by raising the expression of Iduna protein in the pres-ence of OGD/R,hence decreasing the damage to HT22 mouse neurons.

Objective To find the correlation between phosphatidylinositol kinase-3 catalytic subunit A gene(PIK3CA)mutation and pathological features as well as clinical prognosis of breast cancer.Methods The patho-logical data of 181 patients diagnosed with invasive breast cancer from January 2018 to January 2020 were collected.The estrogen receptor(ER),progestogen receptor(PR),human epidermal growth factor receptor-2(HER2),Ki67 were examined by immuno-histochemistry(IHC).Mutation of exon 9 and exon 20 of PIK3CA were examined by quantitative real-time PCR(qPCR).Results Among 181 cases of invasive breast cancer,70 cases had PIK3CA mutation with 31 cases(44.28%)showed exon 9 mutations and 39 cases(55.71%)showed exon 20 mutations.The difference between PIK3CA mutation and their distribution in molecular typing of breast cancer was statistically significant(P＜0.05).The expression of PIK3CA mutation in breast cancer with different Ki67 expression was sig-nificantly different(P＜0.05).There were 34 cases(48.57%)showed PIK3CA mutations in the HR+/HER2 group and 36 cases(51.43%)of non HR+/HER2 group mutations.There was a statistically significant difference in the distribution of PIK3CA mutations between 2 groups(P＜0.05).The death rate of PIK3CA mutation cases was higher than that of PIK3CA wild type cases(P＜0.05).Conclusions PIK3CA mutation is associated with molecular typ-ing,Ki67 increment index and prognosis of breast cancer.Detection of PIK3CA mutation provides potential support to the development of precise treatment of breast cancer patients.

Objective To analyze and compare the clinical efficacy of CalliSpheres drug-eluting micro-spheres and blank microspheres in the treatment of advanced non-small cell lung cancer by bronchial arterial chemoembolization.Methods Fifty patients with advanced non-small cell lung cancer who had failed or relapsed after radiotherapy,chemotherapy,targeting and immunotherapy were collected and treated with super-selective bronchial artery chemoembolization.A retrospective analysis was conducted to compare the tumor response rate and survival between CalliSpheres drug-eluting and blank microspheres.Results The PR,ORR and DCR in the drug-eluted microsphere group were higher than those in the blank microsphere group,and there was a statistical difference in DCR between the two groups 1 month after surgery(χ2 = 4.08,P = 0.04).PD in the drug-eluted microsphere group was lower than that in the blank microsphere group.The CEA,CYF and SCC in the drug-eluted microsphere group after surgery were lower than those in the blank microsphere group,and the CEA,CYF and SCC in the two groups after surgery were lower than those before surgery,and there were statistical differences in CEA and CYF 1 month after surgery between the two groups.The PFS and OS in drug-eluted microsphere group were higher than those in blank microsphere group.Conclusion CalliSpheres drug-eluting microspheres could improve the effective rate of tumor treatment and prolong the survival time more effectively than the blank micro-spheres via arterial chemoembolization,providing reliable clinical practice basis for the treatment of advanced non-small cell lung cancer.

Objective:To explore the diagnostic value of ultrasound elastography (UE) in patients with obese polycystic ovary syndrome (PCOS).Methods:A total of 86 PCOS patients diagnosed in the Second People′s Hospital of Beilun District, Ningbo City from June 2019 to April 2022 were selected as the study group, including 32 patients with body mass inde(BMI) ≥25 kg/m 2 (obese group) and 54 patients with BMI < 25 kg/m 2 (non-obese group). Another 40 women with healthy physical examination and matching age and BMI of the study group were selected as the normal control group. All of them underwent routine vaginal ultrasound and UE examination, and the differences of parameters in each group were compared. The receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic efficacy of relevant parameters for PCOS and obesity PCOS. Results:The number of follicles and ovarian volume in the study group were higher than those in the normal control group, and the PI and RI of ovarian interstitial artery were lower than those in the normal control group: (17.50 ± 3.23) unit vs. (8.15 ± 2.01) unit, (12.97 ± 3.20) ml vs. (5.36 ± 1.82) ml, 0.82 ± 0.14 vs. 0.93 ± 0.20, 0.52 ± 0.12 vs. 0.58 ± 0.10, there were statistical differences ( P<0.05). The number of follicles and ovarian volume in the obese group were significantly higher than those in the non-obese group: (18.98 ± 3.45) unit vs. (16.62 ± 3.17) unit, (15.00 ± 3.15) ml vs. (11.77 ± 2.63) ml, there were statistical differences ( P<0.05); but the PI and RI between the obese group and the non-obese group had no significant differences ( P>0.05). In the study group, 56 cases (65.12%) had type 1 elastic pattern, which was significantly higher than 4 cases (10.00%) in the normal control group ( χ2 = 17.61, P<0.01). The elastic pattern was no significant difference between the obese group and the non-obese group ( P>0.05). The elastic coefficient (B/A ratio) in the study group was higher than in the normal control group:7.86 ± 1.51 vs. 4.53 ± 1.10, there was statistical difference ( P<0.05). The B/A ratio in the obese group was higher than that in the non-obese group and the normal control group: 8.76 ± 1.35 vs. 7.32 ± 1.34, 4.53 ± 1.10, there were statistical differences ( P<0.05). The ROC curve analysis showed that the area under the curve (AUC) of B/A ratio for the diagnosis of PCOS was 0.962, when the cut-off value was 5.56, the corresponding sensitivity and specificity were 93.02% and 87.50%. The AUC of B/A ratio for the diagnosis of obese PCOS was 0.788, when the cut-off value was 8.35, the corresponding sensitivity and specificity were 78.12% and 85.19%. Conclusions:The UE has great diagnostic value for PCOS and certain discrimination performance for obese PCOS.

Background Results of genetic testing for antipsychotic drugs can guide the rational use of drugs in clinical practice and help improve the clinical symptoms of patients with schizophrenia.However,there is currently limited evidence in China regarding the impact of genetic testing results on medication adherence,social function and drug side effects of antipsychotic drug treatment.Objective To explore the improvement of clinical symptoms,medication adherence and social function in patients with schizophrenia under the guidance of antipsychotic drug gene testing results and examine the safety of drug treatment,so as to provide references for ifor precise treatment of schizophrenia patients.Methods Patients with acute schizophrenia who received hospitalization at Dazhou Minkang Hospital from July 2019 to August 2021 as well as met the diagnostic criteria of the International Classification of Diseases,tenth edition(ICD-10)were selected as research subjects(n=144).Based on random number table,subjects were divided into study group and control group,with 72 cases in each group.Control group received drug treatment based on the doctor's clinical experience,while study group received drug treatment based on the results of gene testing for antipsychotic drug.Both treatments lasted for 12 weeks.At baseline as well as 2,4,8 and 12 weeks after treatment,Positive and Negative Syndrome Scale(PANSS),8-item Morisky Medication Adherence Scale(MMAS-8),Social Functional Rating Scale(SFRS)and Treatment Emergent Symptom Scale(TESS)were adopted for assessment.Result Time effect and group effect of the reducing rate of PANSS,MMAS-8 and SFRS scores in the groups were statistically significant(Ftime=95.251,6.650,14.101,Fgroup=38.055,58.175,128.221,P<0.01).The interaction effect of the reduction rates of MMAS-8 scores in two groups was statistically significant(Finteraction=5.837,P<0.01).The group effect and interaction effect of the severity scores of drug side effects and patient pain scores in two groups were statistically significant(Fgroup=7.553,81.533,Finteraction=8.693,9.322,P<0.01).Conclusion In terms of improving clinical symptom relief,medication adherence,social function and drug side effects,medication for patients with schizophrenia guided by genetic testing of antipsychotic drugs may be more effective than that relying on medication based on clinical experience.

Objective To analyze the application of serological test results in the diagnosis and treatment of anti-M-in-duced hemolytic disease of the fetus and newborn(HDFN),and to explore HDFN prevention strategies.Methods The se-rological test results of 12 cases of HDFN caused by anti-M diagnosed in our laboratory from January 2017 to December 2023 were retrospectively analyzed,including blood group identification of mothers and children,serum total bilirubin/hemoglo-bin/antibody titer test,and three hemolysis tests in newborns.Clinical data of the children and mothers were collected,in-cluding pregnancy history,blood transfusion history,prenatal antibody testing,history of intrauterine blood transfusion and gestational week of delivery,and the prognosis of the children was followed up.Results All 12 cases of fetal neonatal he-molytic disease due to anti-M were RhD+MN phenotype newborn born to RhD+NN mother,with maternal-fetal incompati-blility in MN blood groups.In the ABO blood group system,ABO incompatibility between mother and child accounted for 41.7%(5/12).None of the mothers had a history of blood transfusion,and the median titer of the test at 4℃was 32,and the median titer at 37℃was 4.The mothers of 3 cases had a history of multiple intrauterine blood transfusions,with an inci-dence of 25%(3/12).One case had an abnormal first pregnancy,with an incidence of 8.3%(1/12),and seven cases had an abnormal pregnancy with a miscarriage,with an incidence of abnormal pregnancy and birth history of 58.3%(7/12).There were 6 cases of premature labor,with an incidence of 50%(6/12).The mothers in three cases underwent regular ob-stetric examination and the specificity of the antibodies was determined,accounting for 25%(3/12).Twelve children had free antibodies with a median titer of 6 at 4℃and 2 at 37℃.Two children had anti-M antibodies that were not reactive at 37℃,with a negative rate of 16.7%(2/12).The positive rate of DAT and elution test was respectively 8.3%(1/12)and 16.7%(2/12)in the children.The median minimum hemoglobin value was 75 g/L,and all 12 children received blood transfusions.The median peak total bilirubin value was 157.5 μmol/L,and none of them reached the threshold for blood ex-change.The rate of delayed anemia was16.7%(2/12),the postnatal mortality rate was8.3%(1/12),and 11 children was free of growth and neurodevelopmental delay in prognosis.Conclusion Anti-M can cause severe HDFN,which can also oc-cur in primigravida.The intensity of antibody titer does not correlate with the severity of the disease,and it is prone to cause delayed anemia,which should be monitored regularly according to the serological characteristics of anti-M and clinical symp-toms,and should be treated timely.