Objective To investigate the expression level and clinical application value of serum resistin in patients with systemic lu-pus erythematosus(SLE).Methods Forty-five SLE patients visited Nanjing Drum Tower Hospital,Clinical College of Nanjing Uni-versity of Chinese Medicine from January to August 2023 were enrolled in the study.The patients were scored and grouped according to the SLE disease activity index(SLEDAI),with SLEDAI＜9 score in the inactive group(n=32)and SLEDAI≥9 score in the active group(n=13).Thirty-four healthy individuals who underwent physical examination in our hospital were recruited as healthy controls.The clinical data and laboratory related indicators such as urine protein and serum complement C3 levels were collected from SLE pa-tients and healthy controls.Serum resistin levels were detected by enzyme-linked immunosorbent assay(ELISA).The clinical screening value of serum resistin for SLE was evaluated with the receiver operating characteristic(ROC)curve.The correlations of serum resistin levels with different laboratory indicators were determined by Pearson correlation analysis.Results The serum resistin levels in SLE patients([7.64±0.64]ng/mL)were significantly higher than that in healthy controls([2.56±0.43]ng/mL),and the difference was statistically significant(t=6.195,P＜0.01).The serum resistin levels in active SLE patients([10.10±1.45]ng/mL)were significant-ly higher than that in inactive SLE patients([6.64±0.60]ng/mL),and the difference was statistically significant(t=2.632,P＜0.05).The area under the ROC curve(AUCROC)of serum resistin for screening SLE was 0.897.When the cut-off value was 5.893 ng/mL,the sensitivity was 86.67%and the specificity was 82.35%.The serum resistin level in SLE patients was positively correlated with urine protein(r=0.692,P＜0.01),while negatively correlated with serum complement C3(r=-0.354,P＜0.05).Conclusion The expression levels of serum resistin in SLE patients are significantly increased and positively correlated with SLE disease activity and urine protein.Serum resistin may become a novel biomarker for the diagnosis and therapeutic effect assessment of SLE.

Objective To investigate the effhect of ultrasound-guided nerve block on the prognosis of patients undergoing laparoscopic cholecystectomy.Methods A total of 150 cases undergoing elective laparoscopic cholecystectomy in our hospital was used as the research object,and randomly divided into control (n =75) and observation (n =75) groups.The control group used punctured local infiltration analgesia,and observation group used the transversus abdominis plane block (TAP) combined with local infiltration analgesia hole.The postoperative awake time,hemodynamic changes,pain score,and analgesic use were compared between two groups.Results The recovery time was significantly reduced in the observation group (9.7 ± 2) minutes compared to the control group (23.9 ± 2.4) minutes.The additional total dose of analgesics and average additional times were significantly lower in the observation group compared to the control group (P ＜0.05).The pain score were 0.99 ±0.22 after waking,1.05 ±0.14 at the postoperative 6 h,1.13 ±0.23 at the postoperative 24 h,and 1.40 ±0.38 at the postoperative 48 h in the observation group,which was significantly lower than the corresponding 3.99 ± 0.36,3.67 ± 0.43,3.39 ± 0.38,and 3.93 ± 0.20 in the control group (P ＜ 0.05).With the postoperative time,postoperative 48 h in two groups of visual pain scores were significantly higher than that of 24 h after the operation (P ＜ 0.05).The incidence of adverse reactions was no significant difference between observation group (36％) and control group (32％) (P ＞ 0.05).Conclusions Ultrasound-guided nerve block can obviously reduce postoperative pain,reduce drug use frequency and dosage,significantly improve postoperative outcomes,and does not increase the incidence of adverse drug reactions.It is worthy of clinical reference.

Objective To investigate the abnormality of intrahepatic biliary epithelial cells autophagy in primary biliary cirrhosis (PBC) mice,and explore the mechanism of UC-Mesenchymal stem cell (MSCs) in treating PBC.Methods After establishing the PBC model,we divided them into the PBC model group;the UC-MSCs treatment group and the Stattic group [Signal transducer and activator of transcription 3 (STAT3) inhibitor group].Six mice were used as the control group.Liver pathology and the serum pyruvate dehydrogenase complex E2 subunit (PDC-E2) antibody titers were detected.Autophagosome of the intrahepatic biliary epithelial cell was observed by electronic microscope.Protein levels of STAT3/pSTAT3,Beclin-1 were detected by western blot.We cultured human intrahepatic biliary epithelial cells in vitro,and down regulated STAT3.After stimulated by GCDC,we co-cultured them with UC-MSCs,and collected the cells in order to detect LC3 Ⅱ.The measurement data were compared with t test or single factor analysis of variance.Results Compared with the control group,periportal inflammatory cell infiltration and granuloma formation were observed in the PBC group.MSCs treatment decreased the infiltration of inflammatory cells.The level of antiPDC-E2 of the PBC group (107±18) ng/ml was higher than that of the control group (42±6) ng/ml (q=6.326,P＜0.01),MSCs treatment down regulated anti-PDC-E2 level (43±4) ng/ml (q=5.801,P＜0.01).More autophagosomes in the PBC group (5.00±1.29) than the control group (1.75±0.25) were observed (q=4.061,P＞0.05).Western blot showed that the level of Beclin-1 was higher in PBC group (1.80±0.36) than the control group (0.40±0.20) (q =6.757,P＜0.01),MSCs reduced the expression of Beclin-1 (0.86±0.06)(q=4.536,P＜0.05) as well as Stattic (0.72±0.03) (q=5.226,P＜0.05).PBC group had a higher expression level of STAT3 (1.80±0.42) (q=5.730,P＜0.05) and pSTAT3 (2.04±0.29)(q=6.492,P＜0.01) than the control group (0.50±0.05)(0.91±0.14).MSCs treatment decreased the expression of STAT3 (0.51±0.13)(q =5.703,P＜0.01) and pSTAT3 (0.76±0.07) (q =7.388,P＜0.01) in intrahepatic biliary epithelial cells.After down regulated STAT3 of HiBECs,MSCs reduced the expression of LC3 Ⅱ of HiBECs.Conclusion The intrahepatic biliary epithelial cells autophage of PBC mice is abnormal,MSCs can alleviate PBC by down regulating the autophage of intrahepatic biliary epithelial cells via STAT3.

Objective The purpose of this study is to observe the changes of immune cell subsets in lupus mice after umbilical cord mesenchymal stem cells (UC-MSCs) transplantation. Methods B6.MRL-Faslpr lupus mice were randomly divided into the following three groups: the UC-MSCs treated group, the fibroblast like synoviocytes (FLS) treated group and the untreated group. MSC (1×106) or FLS (1×106) were injected into the tail vein of lupus mice respectively. Four weeks after treatment, the spleen index was calculated. The pathological changes of kidney were assessed by HE staining. The frequencies of immune cell subsets in spleen and macrophage in kidney as well as abdominal cavity were analyzed by flow cytometry. Data were analyzed with t test. Results The spleen index of UC-MSCs treated lupus mice [(79 ±9) mg/10 g] and IgG level [(7.5±1.5) mg/ml] were significantly decreased when compared with FLS treated group [(147±23) mg/10 g, t=2.78, P<0.01] [(17.0 ±2.8) mg/ml, t=3.00, P<0.01] and the untreated group [(156 ±16) mg/10 g, t=4.29, P<0.01] [(16.7 ±1.6) mg/ml,t=4.01, P<0.01]. HE staining also showed that the pathological changes of kidney were alleviated after MSCs transplantation. In addition, the frequency of plasma cells in the untreated group [(2.61 2± 0.318)% vs (0.306±0.017)%, t=7.22, P<0.01] and the FLS treated group [(2.412±0.297)% vs (0.306±0.017)%, t=7.07, P<0.01] were markedly higher than MSCs treatment [(0.306 ±0.017)%]. Moreover, the frequency of CD25+Foxp3+/CD4+Treg in the MSCs treated group [(15.08±0.81)%] was significantly increased compared with the untreated group [(8.02 ±0.47)%, t=7.45, P<0.01] and FLS treated group [(8.80 ±0.23)%, t=7.39, P<0.01]. MSCs treatment resulted in a decrease in CXCR5+PD1+/CD4+Tfh and IFNγ+/CD4+Th1 subset, compared with the untreated group [(14.3±1.5)%vs (31.5±3.3)%, t=5.25, P<0.01] [(1.78±0.27)% vs (5.93±1.56)%, t=2.60, P<0.05] and the FLS treated group [(14.3±1.5)%vs (28.8±2.2)%, t=5.49, P<0.01] [(1.78±0.27)%vs (4.88±0.81)%, t=3.61, P<0.01]. The frequency of macrophage in kidney of the MSCs treated group [(3.52 ±0.37)%] was markedly increased compared with the untreated group[(1.58±0.29)%, t=3.25, P<0.01], while neither the IL4+/CD4+Th2 subset nor the IL17+/CD4+Th17subset and the frequency of macrophage in abdominal cavity showed significant changes in the three groups. Conclusion These findings suggest that the therapeutic effects of MSCs on lupus mice may mediate through increasing the frequency of spleen Treg and renal macrophage and decreasing the frequency of Tfh, Th1 and plasma cells.

Objective To explore the effect of flipped classroom on basic nursing operation training for undergraduate nursing students. Methods One hundred full-time nursing undergraduate students now conducting clinical internship in our hospital were randomized equally into experiment group and control group. The control group was treated with traditional teaching model in three nursing operation modules including intravenous infusion , venous blood collection and intramuscular injection while the teaching methodology of flipped classroom was used in the experiment group. The two groups were compared in terms of performances in theoretical knowledge and skills and the evaluation from the students. Results The achievements of the experiment group was significantly better than those of the control group in respect of achievements in the examinations of three theoretical knowledge and nursing operation skills (P<0.01). More than 88.00%nursing students agreed on the teaching mode in the aspects of learning attitude and knowledge mastering, more than 90.00%nursing students accepted this teaching mode in the aspect of ability and 96.00%of them would like to adopt this mode. Conclusions The methodology of flipped classroom is effective in cultivating the habit of nursing students′autonomous learning. It also stimulates the nursing students′interest in learning and improves the quality of teaching.

OBJECTIVETo explore the molecular mechanism underlying the DEL phenotype among RhD negative ethnic Han individuals from Jiangsu, China.

METHODSThe DEL phenotype was determined by an adsorption elution test among 57 RhD negative blood donors. The Rh C, c, E, and e phenotypes were detected by a tube method. PCR with sequence-specific primering (PCR-SSP) assay was used to determine the RHCE genotypes. The RHD gene of the DEL individuals were amplified with polymerase chain reaction and subjected to Sanger sequencing analysis.

RESULTSAmong the 57 RhD negative donors, 10 (17.54%) were determined as having the DEL phenotype. The major RhCE phenotypes for DEL and RhD negative cases were RhCcee (80.0%) and Rhccee (61.7%), respectively. All RHD gene sequences of the 10 individuals have harbored a G>A mutation at position 1227 of exon 9.

CONCLUSIONA proportion of RhD negative individuals determined by routine serological method are actually DEL with RHD gene mutations. RHD *1227A is the most prevalent DEL genotype among ethnic Han Chinese from Jiangsu. Further research on the phenotype and underlying molecular mechanism of DEL is important for blood transfusion.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Exons ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics

Objective To explore the preventive effect of early umbilical cord mesenchymal stem cells (UC-MSCs) transplantation on MRL/lpr mice and the underly mechanisms.Methods Fourteen 10-week-old MRL/lpr mice were labeled and numbered.They were randomly divided into 2 groups by using random number table and injected with 1 ×106 UC-MSCs or PBS via tail vein respectively.Proteinuria was measured with Bradford method every 4 weeks.All mice were sacrificed at the age of 28 weeks, with the level of serum antidsDNA antibody and IL-17 detected by enzyme linked immunosorbent assay (ELISA).Splenic Th17 cells, as well as regulatory T cells (Treg) were examined by flow cytometry.Data were analyzed with t test and Pearson's correlation test.Results The onset of proteinuria was delayed for 4 weeks in UC-MSC-treated group compared with that in the control group.At the age of 28 weeks, the 24 hour proteinuria [(1.78±0.17) mg vs (4.77±0.98)mg, t=2.99, P＜0.05] and the spleen weight [(0.149±0.009) g vs (0.273±0.052) g, t=2.33, P＜0.05] in UC-MSCtreated group were significantly lower than those in the control group.There was also a trend of the decline of serum anti-dsDNA antibody and IL-17 level after UC-MSCs transplantation.Compared with those in the control group, both the percentage and the absolute number of Th17 cells were significantly decreased in UC-MSC-treated group [(0.90±0.19)％ vs (2.81±0.50)％, t=3.54, P＜0.01 and (3.7±0.8)×105 vs (19.3±3.7)×105, t=4.12,P＜0.01].Meanwhile, the percentage of Treg elevated after UC-MSCs treatment.The ratio of Th17/Treg was significantly lower in UC-MSC-treated group than that in the control group (0.11±0.03 vs 0.50±0.09, t=4.23,P＜0.01).Both the ratio of Th17/Treg (r=0.73, P＜0.01;r=0.59, P＜0.05) and serum IL-17 level (r=0.78, P＜0.01;r=0.56, P＜0.05) was positively correlated with the level of 24 hour proteinuria and anti-dsDNA antibody respectively in MRL/lpr mice.Conclusion Early UC-MSCs transplantation helps to delay disease onset and ameliorate disease progression in MRL/lpr mice, which may act through the modulation of Th17/Treg balance.

Objective Whether the bone marrow cells (BMC) derived from systemic lupus erythematosus (SLE) could transmit autoimmune disease was studied for the purpose of clarifying the role of BMC in SLE pathogenesis.The effects of bone marrow mesenchymal stem cells (MSC) from SLE and control mice on the SLE BMC-induced symptoms were compared to elucidate the role of MSC in SLE.Methods Six-week-old B6.MRL-Fas mice were randomly divided into 3 groups.One group was transplanted with BMC from the 30-week-old B6.MRL-Faslg mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Faslpr mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Faslg mice and bone marrow MSC from the age-matched C57BL/6 mice.Before transplantation,the recipient mice received irradiation by an X-ray source.The levels of serum antinuclear antibody (ANA) and proteinuria were measured with enzyme linked immunosorbent assay (ELISA) and Bradford method every 4 weeks,respectively.The survival rate was recorded.All mice were sacrificed 18 weeks later.Splenic plasma cells,Th1,Th2 and Th17 cells were measured by flow cytometry.Statistical analyses were performed using the independent t test and ANOVA.Results Eight weeks after transplantation,ANA was positive in all the recipient mice.However,there was no significant difference between the three groups (P＞0.05).No proteinuria was observed in all the recipient mice.The mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Fasr mice showed an elevated trend of the percentages of splenic plasma cells,Th1,Th2 and Th17 cells compared with the other two groups,plasma cells [(1.05±0.16)％,(0.58±0.11)％,t=2.53,P＞0.05;(1.05±0.16)％,(0.71±0.18)％,t=1.45,P＞0.05],Th1 cells [(6.6±2.2)％,(5.7±1.0)％,t=0.38,P＞0.05;(6.6±2.2)％,(4.0±1.7)％,t=0.96,P＞0.05],Th2 cells [(3.3±0.4)％,(2.1±0.6)％,t=1.76,P＞0.05;(3.3±0.4)％,(2.2±0.6)％,t=1.51,P＞0.05],Th17 cells [(2.30±0.71)％,(1.31±0.31)％,t=1.27,P＞0.05;(2.30±0.71)％,(1.12±0.27)％,t=1.67,P＞0.05].However,there was no significant difference between the groups.The survival rate of the three groups was 43％,43％ and 80％ respectively.And the survival rate of the mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched C57BL/6 mice was significantly higher than those of the other groups.Conclusion Our results indicate that BMC from SLE can transmit autoimmune disease.The bone marrow MSC can not prevent lupus-like presentations induced by BMC from SLE.Transplantation of bone marrow MSC from C57BL/6 mice can significantly elevate the survival rate.

Objective To study the blood concentration of hydroxychloroquine in patients with systemic lupus erythematosus (SLE) treated with different doses, and analyze the relationship between blood concentration of hydroxychloroquine and disease activity, and evaluate its safety.Methods Forty SLE patients were randomly divided into 2 groups, each group contained 20 cases.The patients in group A were treated with hydroxychloroquine (0.4 g, qd), while patients in group B were treated with hydroxychloroquine (0.2 g, qd).The treatment lasted more than six months in every patient.The blood concentrations of hydroxychloro-quine were detected by high performance liquid chromatography.The clinical and laboratory indices were collected.The systemic lupus erythematosus disease activity index (SLEDAI) score was recorded.The doses and varieties of combined hormone, immunosuppressant were recorded.The correlation of blood concentration of hydroxychloroquine and disease activity was analyzed.The significance was determined by Student's t test and Pearson correlation analysis.Results In SLE patients, the average blood concentration of hydro-xychloroquine was (402±190) ng/ml in group A and (150±60) ng/ml in group B (t=7.471, P＜0.01).The disease activities of patients in the two groups showed no significant difference (t=-0.172, P＞0.05).The platelet counts of patients in group A were significantly higher than those in group B[(188±88)×109/L vs (158 ±87) ×109/L] (t=4.375, P＜0.05).However, the other laboratory parameters showed no significant difference between the two groups (P＞0.05).Conclusion The results of this study indicate that the blood concentrations of hydroxy-chloroquine are significantly different in different dosages.The high dose of hydroxy-chloroquine is related to high platelet number in lupus patients.These findings suggest that hydroxychloroquine is safe and effective for SLE patients.

OBJECTIVETo evaluate the role of germline mutations of TP53 gene among a Chinese population with high risk for breast cancer.

METHODSA total of 81 BRCA-negative breast cancer probands from cancer families were analyzed using targeted capture and next-generation sequencing. Candidate mutations were verified with Sanger sequencing. Co-segregation analyses were carried out to explore the likely pathogenicity of the mutation.

RESULTSOf the 81 BRCA-negative patients, 3 exonic mutations in the TP53 gene were identified in 3 breast cancer patients. Of these, 2 mutations were previously reported and 1 was novel. One family with TP53 mutation has met the criteria for Li-Fraumeni syndrome (LFS) and accounted for 9.1% of all families who fulfilled the diagnostic criteria for LFS. Two of the carriers were diagnosed with breast cancer under the age of 30, and have accounted for 11.8% (2/17) of all very young (≤30 years) breast cancer patients in our study.

CONCLUSIONThe TP53 germline mutation is more common in Chinese population with a high risk for breast cancer than previously thought. TP53 gene mutation screening should be considered particularly for patients with a family history of LFS and very young age of onset.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Breast Neoplasms ; ethnology ; genetics ; China ; DNA Mutational Analysis ; Exons ; Family Health ; Female ; Genetic Predisposition to Disease ; ethnology ; genetics ; Germ-Line Mutation ; Heterozygote ; Humans ; Li-Fraumeni Syndrome ; ethnology ; genetics ; Male ; Middle Aged ; Pedigree ; Risk Factors ; Tumor Suppressor Protein p53 ; genetics ; Young Adult