OBJECTIVE To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib. METHODS Using human liver cancer cell Huh7 as subject， the lenvatinib-resist cell model （Huh7-LR） was generated by the low-dose gradient method combined with long-term administration. The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry. Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1， autophagic adapter protein sequestosome 1 （p62）， microtubule-associated protein 1 light chain 3 （LC3） and autophagic level. Furthermore， an autophagy activation model was constructed by cell starvation， the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection， and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry. RESULTS Compared with parental cells， the drug resistance index of Huh7-LR cells was 6.2； protein expression of p62 was increased significantly， while apoptotic rate， protein expression of Beclin-1 and LC3Ⅱ/ LC3Ⅰ ratio were all reduced significantly （P＜0.05 or P＜0.01）； the level of autophagy was decreased to some extent. Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells （P＜0.05） and autophagy level， and significantly increase its apoptotic rate （P＜0.05）. CONCLUSIONS Autophagy is involved in lenvatinib resistance， and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.

Objective:To analyse the clinical significance of selective single embryo transfer by time-lapse mo-nitoring(TLM)or conventional morphology assessment(CMA)in vitro fertilization/intracytoplasmic sperm in-jection and embryo transfer(IVF/ICSI-ET),and to initially explore the predictive value of Raman spectral analy-sis of embryo culture medium for clinical pregnancy rate.Methods:The study is a prospective randomized con-trolled clinical trial.We assigned 139 patients treated with IVF/ICSI-ET in Reproductive and Genetics Center of Suzhou Municipal Hospital from April 2019 to July 2020,which were randomly assigned to either the CMA or the TLM group.We performed selective single-embryo transfer(fresh cycle and FET)after selecting the optimal em-bryos with TLM or CMA respectively.If the patient's first embryo transfer was unsuccessful,a second one would be performed to compare the differences in the cumulative live birth rate of embryo transfer and other pregnancy outcomes between the two groups.Meanwhile,we collected 15 μl of embryo culture medium at day 3 after IVF/ISCI fertilization for Raman spectroscopy analysis.Results:There were no differences in cumulative live birth,cu-mulative clinical pregnancy,cumulative premature birth,cumulative early spontaneous abortion,cumulative ectopic pregnancy and LGA or SGA between TLM and CMA groups(P＞0.05).The Neonatal sex ratio in the TLM group was lower than that in the CMA group,but the difference was not significant(P＞0.05).Raman spectros-copy analysis of embryo culture medium predicted the clinical pregnancy rate with 67.21％accuracy.Conclu-sions:In young women with a good ovarian reserve,the advantage of using TLM to evaluate embryos is not obvi-ous,so we should remain vigilant that embryo selection based on morphokinetic parameters may affect the sex ratio.Raman spectroscopic analysis of embryo culture medium is not yet able to effectively predict the planting ability of embryos.

Objective:To study the feasibility of magnetic resonance imaging(MRI)technique with amide proton transfer(APT)in predicting the prognosis of cerebral stroke.Methods:A total of 71 patients with acute cerebral stroke who admitted to the Nanjing First Hospital,Nanjing Medical University from September 2022 to May 2023 were selected.All of them underwent the test of National Institute of Health Stroke Scale(NIHSS),and received the MRI examination with chemical exchange saturation transfer(CEST).According to the modified Rankin scale(mRS)values of 1-month follow-up,they were divided into favorable recovery group(mRS<2,44 cases)and poor group(mRS≥2,27 cases).The asymmetric magnetization transfer ratio(MTRasym)image(APT)was obtained by analyzing data with special software.And then,the difference(△APTw)of APT values between ischemic zone and contralateral normal tissue was further calculated.The △APTw values of two groups were compared and analyzed,and the Pearson correlation analysis was adopted to analyze the correlation among △APTw,NIHSS and mRS.The receiver operating characteristics(ROC)curve was drawn,and the area under curve(AUC)of ROC curve was calculated.Results:There were significant positive correlations among △APTw,NIHSS and mRS scores(R2=0.659,0.522,P<0.001),and the differences of △APTW,NIHSS and mRS scores between the favorable recovery group and poor group were significant(t=5.73,6.36,13.92,P<0.05),respectively.The AUC value was 0.886,and the sensitivity and specificity of prediction were respectively 77.8%and 95.5%.The positive and negative predictive values were respectively 91.3%and 87.5%.Conclusion:APT imaging technique has feasibility in predicting the prognosis of acute cerebral ischemic stroke.

Abstract: Objective To clarify the long-term evolution of hepatitis B virus (HBV) quasi-species in HBsAg asymptomatic carriers in Long'an county, Guangxi. Methods ELISA was used to detect serological markers of HBV. Viral loads were measured by real time PCR. HBV DNA was extracted from serum by kits. The whole HBV genome was amplified using nested PCR and amplicons were sequenced by next-generation sequencing (NGS). These sequences from NGS were analyzed by the software like Mega. Results Serum samples were collected from 9 HBsAg asymptomatic carriers in Longan County，Guangxi at 4 different time points in 2004, 2007, 2013, 2019 or 2020. A total of 23 serum samples and 309 full-length gene quasi-species sequences were obtained, with an average amount of (0.18±0.07) G sequencing data for each sample. Genotype of 55.54%(5/9) the studied subjects underwent genotype conversion during the long-term evolution process of HBV quasi-species, and the genotyping results of the phylogenetic tree in the PreS/S region are in perfect agreement with the results of the whole genome analysis; recombinant B/C, I/C were found; the Sn ranged from 0 to 0.37 and the genetic diversity ranged from 0 to 0.11, respectively. A total of 21 special single nucleotide/amino acid mutations (7 in the S region, 2 in the X region, 3 in the PreC region and 9 in the BCP region) and 6 deletion mutations were detected, multiple mutations were found and no drug resistant mutations were found; 77.8%(7/9) of the HBV strains carried by the subjects in 2004 had double mutations at nt1 762(A→T) and 1 764(G→A) and a stop mutation at nt1 896(G→A); HBV mutations can be restored from the mutant type to the wild type and (or) vice versa without antiviral drug pressure, and The evolution rate of HBV genome was 2.03×10-5~3.50×10-3.Conclusion HBV genotype, recombinants, genetic complexity and diversity of HBV quasi-species can change over time during in natural infection. The transformation between HBV mutation type and wild type reduces the value of predicting clinical outcomes by genetic types and related mutations to some extent. The HBV genome evolution rate of asymptomatic carriers of HBsAg in Long'an County is very high.

Immunoglobulin (IgG) glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases, including systemic lupus erythematosus (SLE), thus underlining the pathogenic role of glycosylation aberration in autoimmunity. This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy. Relative to that in serum samples from the control cohort, IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages (from preconception to the third trimester of pregnancy) and was significantly associated with lupus activity and fetal loss during lupus pregnancy. The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation. The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells (pDCs). RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase (SYK) signaling pathway significantly differed between IgG- and deSia-IgG-treated pDCs. This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG. Finally, the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG. Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.
Humans ; Pregnancy ; Female ; Lupus Erythematosus, Systemic/pathology* ; Signal Transduction ; N-Acetylneuraminic Acid/metabolism* ; Immunoglobulin G ; Dendritic Cells/pathology*

OBJECTIVE To investigate the mechanism of glucose -6-phosphate dehydrogenase （G6PD）-induced sorafenib - resistance in hepatocarcinoma cell based on phoshorylated 3-kinase/protein kinase B （PI3K/Akt）signaling pathway . METHODS Cell lines including hepatocarcinoma cell Huh 7，sorafenib-resistant cell Huh 7-SR，G6PD overexpressed cell Huh 7-G6PD and its control cell Huh 7-CT，and compounds including G 6PD inhibitor （6-Aminonicotinamide，6AN）and sorafenib were used as objects or intervention drugs in these research . CCK8 assay was applied to evaluate cell viability . The protein levels of G 6PD and the phosphorylation levels of PI 3K/Akt signaling pathway were detected by Western blot . Flow cytometry was utilized to investigate cell apoptosis. RESULTS Compared with Huh 7 cells，the protein level of G 6PD was significantly increased in Huh 7-SR cells （P＜ 0.05）. The combination of 6AN and sorafenib reduced cell viability of Huh 7-SR cells （P＜0.01）. However，compared with Huh 7- CT，increased cell viability and decreased cell apoptosis rate were observed in Huh 7-G6PD cells while cells were treated with sorafenib（P＜0.01）. Mechanistically，the phosphorylation levels of PI 3K and Akt were significantly decreased in Huh 7-SR cells that were treated with 6AN（P＜0.05）. Moreover，under the condition of no drug intervention ，the phosphorylation levels of PI 3K and Akt were significantly elevated in Huh 7-G6PD cells when compared with Huh 7-CT（P＜0.01）. CONCLUSION G6PD could induce sorafenib -resistance in hepatocarcinoma cell by activating PI 3K/Akt signaling pathway .

Objective:To explore the effect of midwife-led specialist outpatient weight management on the delivery outcomes of pregnant women with scarred uterus.Methods:From January 2020 to January 2021, convenience sampling was used to select 120 pregnant women with singleton scarred uterus in the First Affiliated Hospital of Wenzhou Medical University as the research object. Pregnant women were divided into the control group and the observation group by random number table method, 60 cases in each group. The control group received routine antenatal outpatient examination and guidance, while the observation group received midwife-led specialist outpatient weight management on the basis of the control group. The vaginal delivery rate, cesarean section rate, 24-hour postpartum hemorrhage, anxiety and newborn birth weight were compared between the two groups.Results:The vaginal delivery rate of pregnant women in the observation group was higher than that in the control group, and the cesarean section rate, 24-hour postpartum hemorrhage, anxiety and newborn birth weight were lower than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Midwife-led specialist outpatient weight management can effectively control the weight gain of pregnant women with scarred uterus during pregnancy, improve delivery outcomes, reduce postpartum hemorrhage and maternal anxiety, and help control the birth weight of newborns within the ideal range, which is worthy of clinical application.

Objective To investigate the correlation between the changes of serum neuron specific enolase(NSE) and hypersensitive C-reactive protein (hs-CRP) levels and the degree of neurological deficit (NIHSS)score in patients with cerebral infarction.Methods From January 2017 to January 2019,63 patients with cerebral infarction admitted to Lishui Central Hospital were selected.According to NIHSS score,they were divided into 13 mild cases,30 moderate cases and 20 severe cases.According to infarction area,they were divided into large area group(16 cases),small area group (27 cases) and lacunar infarction group (20 cases).Another 60 cases underwent health examination in our hospital from January 2017 to January 2019 were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of NSE,and immunoturbidimetric assay was used to determine the content of hs-CRP.The changes of serum NSE and hs-CRP levels in the cerebral infarction group and control group,serum NSE,hs-CRP levels and NIHSS scores in different severity and infarction area,and the correlation between serum NSE and hs-CRP changes and NIHSS scores were compared.Results The serum levels of NSE [(21.34 ± 3.27) ng/mL] and hs-CRP [(10.48 ± 2.14) mg/L] in the cerebral infarction group were significantly higher than those in the control group [(6.23 ± 1.08) ng/mL,(2.83 ± 0.46) mg/L] (t =34.061,27.095,all P ＜ 0.05).The serum levels of NSE [(26.98 ± 3.64) ng/mL],hs-CRP [(15.36 ± 2.57) mg/L] and NIHSS score[(38.49 ±3.25) points] in the severe group were higher than those in the moderate group and mild group,which in the moderate group [(20.98 ± 3.21) ng/mL,(10.25 ± 2.09) mg/L and (22.18 ± 3.48) points]were higher than those in the mild group [(12.64 ± 2.78) ng/mL,(5.47 ± 1.40) mg/L and (7.38 ± 2.56)],the differences were statistically significant (F =14.975,9.132,15.873,all P ＜ 0.05).The serum levels of NSE[(25.43 ± 3.35) ng/mL],hs-CRP [(16.54 ± 2.71) mg/L] and NIHSS score [(37.34 ± 3.75) points] in the large area group were higher than those in the small area group and lacunar infarction group,which in the small area group [(21.67 ± 3.12) ng/mL,(10.86 ± 2.21) mg/L and (21.25 ± 3.26) points] were higher than those in the lacunar infarction group [(13.45 ± 2.97) ng/mL,(4.79 ± 1.35) mg/L and (8.49 ± 2.15) points],the differences were statistically significant (F =13.241,9.893,17.482,all P ＜ 0.05).The serum levels of NSE and hs-CRP were positively correlated with NIHSS score (r =0.829,0.713,all P ＜ 0.05).Conclusion The levels of serum NSE and hs-CRP in patients with cerebral infarction increase with the progression of the disease,and there is a linear positive correlation between NSE and hs-CRP and NIHSS score.It is considered that NSE and hs-CRP are of great value in evaluating the degree of neurological impairment,the severity of the disease and the size of the infarct.

Objective:To explore the effect of laparoscopic surgery on postoperative rehabilitation and stress response of patients with colorectal cancer.Methods:From January 2018 to September 2019, 94 patients with colorectal cancer were selected in the General Hospital of Marine Police of the Chinese People's Armed Police Force.They were divided into observation group (47 cases) and control group (47 cases) according to the random digital table method.The observation group was treated with laparoscopic surgery, while the control group was treated with open surgery.The changes of perioperative indicators, postoperative rehabilitation indicators, postoperative complications and 24-hour stress response indicators were compared between the two groups.Results:The time of lymph node dissection and operation had no statistically significant differences between the two groups (all P>0.05). The amount of bleeding of the observation group [(153.61±27.58)mL] was less than that of the control group [(271.32±23.25)mL] ( t=22.371, P<0.05). In the observation group, the exhaust time [(43.27±6.18)h] and the recovery time of bowel sounds [(20.31±3.28)h] were faster than those in the control group [(67.38±9.91)h and (34.54±5.17)h] ( t=14.153, 15.934, all P<0.05). The incidence of postoperative complications in the observation group (4.26%) was lower than that in the control group (21.28%) (χ 2=6.114, P<0.05). The serum levels of norepinephrine (NE) [(92.13±10.45)μg/L], C-reactive protein (CRP) [(17.38±3.01)mg/L] and Cor [(137.87±23.15)μg/L] in the observation group were lower than those in the control group [(136.27±15.42)μg/L, (26.73±2.34)mg/L and (185.42±27.39)μg/L] ( t=16.245, 16.813, 9.090, all P<0.05). Conclusion:Laparoscopic radical resection of colorectal cancer has good effect on patients with colorectal cancer.It can speed up postoperative rehabilitation and reduce stress response.It is worthy of popularizing.

Objective: To compare the clinical application of three-dimensional laparoscopic pancreatoduodenectomy (3D-LPD) with that of two-dimensional laparoscopic pancreatoduodenectomy (2D-LPD), and to explore the safety and feasibility of 3D-LPD. Methods: A retrospective analysis was made from the data of 45 patients with 3D-LPD and 45 patients with 2D-LPD who underwent total laparoscopic pancreatoduodenectomy from March 2017 to August 2018 at Department of Hepato-Pancreato-Biliary Surgery, the First People′s Hospital of Changzhou.The differences of intraoperative conditions, postoperative complications and postoperative pathological findings between the two methods were compared.Measurement data were compared with independent sample t-test, enumeration data were statistically analyzed with Chi-square test or Fisher exact probability. Results: The operation time of 3D-LPD group was shorter than that of 2D-LPD group ((335±95) min vs. (419±113) min, t=-3.817, P=0.000), which mainly showed that the time of digestive tract reconstruction was reduced ((92±26) min vs. (131±46) min, t=-4.951, P=0.000). The intraoperative blood loss in the 3D-LPD group was significantly less than that in the 2D-LPD group ((242±124) ml vs. (350±176) ml, t=-3.365, P=0.001), and the perioperative blood transfusion in the 3D-LPD group was significantly less than that in the 2D-LPD group (χ²=4.444, P=0.035). Postoperative hospitalization days and ICU stay time were not significantly different between the two groups(both P>0.05). Postoperative complications such as pancreatic fistula, biliary fistula, postoperative bleeding, gastric emptying disorders, abdominal infection, were not significantly different between the two groups(all P>0.05). Conclusions The operation time of 3D-LPD is shorter than that of 2D-LPD, and the amount of bleeding is less. Short-term clinical data showed that, 3D-LPD is effective, safe and worth popularizing.