Objective To explore the feasibility of using cell-free DNA(cfDNA)in blastocyst fluid for preimplantation embryo aneuploidy.Methods A total of 17 patients undergoing assisted reproductive technology at Taizhou Hospital of Zhejiang from 2023 to 2024.A total of 38 discarded blastocysts were collected.Whole-genome amplification and next-generation sequencing were performed on both blastocyst fluid cfDNA and trophoblast cells.Using the trophoblast cells as reference,the amplification success rates of blastocyst fluid cfDNA were compared.The consistency,specificity,and sensitivity of the detection results were evaluated.Results The success rate of cfDNA amplification in the blastocyst fluid was 78.95％,and that of the trophoblast cells was 90.61％,with a consistency of 57.1％.The sensitivity of cfDNA amplification in the blastocyst fluid was 100％,the specificity was 66.7％,the positive likelihood ratio was 3,and the negative likelihood ratio was 0.Conclusion Blastocyst fluid cfDNA screening can be used as a screening method to exclude chromosomal embryos aneuploidy and can be used for preimplantation embryo selection.

This research aimed to comprehensively understand the prevalence of mutation in drug-resistant molecular markers of imported Plasmodium vivax in Chongqing,the Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o and Pvk12 genes of Plasmodium vivax were systematically analyzed.Blood samples were collected from imported Plasmodium vivax-infected patients in Chongqing between 2011 and 2022.The Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o,and Pvk12 genes were amplified and then sequenced to precisely evaluate gene mutations.Bioinformatics methods were employed to conduct in depth analysis of the mutation prevalence.Regarding the Pvdhfr gene,mutations at codons 50,57,58,61,99,117,and 199 were detected in 2.9%,23.5%,76.4%,23.53%,2.9%,82.3%,and 5.88%of the samples,respectively.The double-mutant haplotype S58R/S117N was the most prevalent,accounting for 50%,followed by the quadruple-mutant haplotype F57L/S58R/T61M/S117T,which accounted for 11.76%.Among the four types of tandem-repeat variations of Pvdhfr,the wild-type was the most common,and the insertion type was a novel discovery in this study.For the Pvdhps gene,the prevalence among mutation genotypes was relatively low.The single-mutant genotype was dominant,constituting 27.03%.The prevalence of Pvmdr1 mutations at codons 958 and 1076 was 100%and 89.19%,respectively.Among the 37 successfully sequenced samples,K10 insertion was detected in only 8 cases(22.22%).Notably,no non-synonymous mutations of Pvk12 were identified in this study.The cases in this study were imported from various countries of origin.Novel tandem-repeat variation tyres of Pvdhfr and new mutation sites of Pvdhps were identifide,thus enriching the mutation information of imported Plasmodium vivax resistance molecular markers in China.

Objective To systematically review the efficacy and safety of tegoprazan in the treatment of gastroesophageal reflux disease(GERD).Methods PubMed,Embase,Web of Science,Cochrane Library,SinoMed,CNKI,VIP,and WanFang Data databases were electronically searched to collect randomized controlled trials(RCTs)on tegoprazan for GERD treatment from inception to January 1,2025.Two researchers independently screened the literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was performed using RevMan 5.4 software and Stata 16 software.Results A total of 7 RCTs were included,involving 1,329 patients.The results of Meta-analysis showed that there was no statistically significant difference in the overall effective rate between the tegoprazan group and the control group(placebo or proton pump inhibitors)[RR=1.08,95%CI(0.99,1.17),P=0.07].There was also no statistically significant difference in the cure rate between the tegoprazan group and the control group(proton pump inhibitors)[RR=0.99,95%CI(0.96,1.02),P=0.53].Comparing the incidence of treatment-emergent adverse events(TEAE)and serious adverse events(SAE)during treatment between the tegoprazan group and the control group,no statistically significant differences were found[TEAE:RR=0.90,95%CI(0.62,1.32),P=0.60;SAE:RR=0.61,95%CI(0.26,1.48),P=0.28].In terms of specific adverse event,the incidence of abnormal liver function was significantly higher in the tegoprazan group compared to the control group[RR=7.60,95%CI(1.40,42.27),P=0.02],while the incidence of other adverse reactions showed no significant differences(P>0.05).Conclusion Tegoprazan has relatively good overall efficacy and safety in the treatment of GERD,and its efficacy is similar to that of proton pump inhibitors,which can be used as an alternative treatment for proton pump inhibitors.Due to the limited quality and quantity of the included studies,more high-quality RCTs are needed to verify the above conclusion.

Objective To explore the feasibility of using cell-free DNA(cfDNA)in blastocyst fluid for preimplantation embryo aneuploidy.Methods A total of 17 patients undergoing assisted reproductive technology at Taizhou Hospital of Zhejiang from 2023 to 2024.A total of 38 discarded blastocysts were collected.Whole-genome amplification and next-generation sequencing were performed on both blastocyst fluid cfDNA and trophoblast cells.Using the trophoblast cells as reference,the amplification success rates of blastocyst fluid cfDNA were compared.The consistency,specificity,and sensitivity of the detection results were evaluated.Results The success rate of cfDNA amplification in the blastocyst fluid was 78.95％,and that of the trophoblast cells was 90.61％,with a consistency of 57.1％.The sensitivity of cfDNA amplification in the blastocyst fluid was 100％,the specificity was 66.7％,the positive likelihood ratio was 3,and the negative likelihood ratio was 0.Conclusion Blastocyst fluid cfDNA screening can be used as a screening method to exclude chromosomal embryos aneuploidy and can be used for preimplantation embryo selection.

This research aimed to comprehensively understand the prevalence of mutation in drug-resistant molecular markers of imported Plasmodium vivax in Chongqing,the Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o and Pvk12 genes of Plasmodium vivax were systematically analyzed.Blood samples were collected from imported Plasmodium vivax-infected patients in Chongqing between 2011 and 2022.The Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o,and Pvk12 genes were amplified and then sequenced to precisely evaluate gene mutations.Bioinformatics methods were employed to conduct in depth analysis of the mutation prevalence.Regarding the Pvdhfr gene,mutations at codons 50,57,58,61,99,117,and 199 were detected in 2.9%,23.5%,76.4%,23.53%,2.9%,82.3%,and 5.88%of the samples,respectively.The double-mutant haplotype S58R/S117N was the most prevalent,accounting for 50%,followed by the quadruple-mutant haplotype F57L/S58R/T61M/S117T,which accounted for 11.76%.Among the four types of tandem-repeat variations of Pvdhfr,the wild-type was the most common,and the insertion type was a novel discovery in this study.For the Pvdhps gene,the prevalence among mutation genotypes was relatively low.The single-mutant genotype was dominant,constituting 27.03%.The prevalence of Pvmdr1 mutations at codons 958 and 1076 was 100%and 89.19%,respectively.Among the 37 successfully sequenced samples,K10 insertion was detected in only 8 cases(22.22%).Notably,no non-synonymous mutations of Pvk12 were identified in this study.The cases in this study were imported from various countries of origin.Novel tandem-repeat variation tyres of Pvdhfr and new mutation sites of Pvdhps were identifide,thus enriching the mutation information of imported Plasmodium vivax resistance molecular markers in China.

Objective To systematically review the efficacy and safety of tegoprazan in the treatment of gastroesophageal reflux disease(GERD).Methods PubMed,Embase,Web of Science,Cochrane Library,SinoMed,CNKI,VIP,and WanFang Data databases were electronically searched to collect randomized controlled trials(RCTs)on tegoprazan for GERD treatment from inception to January 1,2025.Two researchers independently screened the literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was performed using RevMan 5.4 software and Stata 16 software.Results A total of 7 RCTs were included,involving 1,329 patients.The results of Meta-analysis showed that there was no statistically significant difference in the overall effective rate between the tegoprazan group and the control group(placebo or proton pump inhibitors)[RR=1.08,95%CI(0.99,1.17),P=0.07].There was also no statistically significant difference in the cure rate between the tegoprazan group and the control group(proton pump inhibitors)[RR=0.99,95%CI(0.96,1.02),P=0.53].Comparing the incidence of treatment-emergent adverse events(TEAE)and serious adverse events(SAE)during treatment between the tegoprazan group and the control group,no statistically significant differences were found[TEAE:RR=0.90,95%CI(0.62,1.32),P=0.60;SAE:RR=0.61,95%CI(0.26,1.48),P=0.28].In terms of specific adverse event,the incidence of abnormal liver function was significantly higher in the tegoprazan group compared to the control group[RR=7.60,95%CI(1.40,42.27),P=0.02],while the incidence of other adverse reactions showed no significant differences(P>0.05).Conclusion Tegoprazan has relatively good overall efficacy and safety in the treatment of GERD,and its efficacy is similar to that of proton pump inhibitors,which can be used as an alternative treatment for proton pump inhibitors.Due to the limited quality and quantity of the included studies,more high-quality RCTs are needed to verify the above conclusion.

Objective To investigate the value of serum cell cycle protein-dependent kinase 1(CDK1)and aurora kinase A(AURKA)in the diagnosis of patients with hepatitis B virus-related hepatocellular carcinoma(HBV-HCC).Methods A total of 50 HBV-HCC patients,50 patients with hepatitis B virus-related liver cirrhosis(HBV-LC),and 50 chronic hepatitis B(CHB)patients who were hospitalized in Department of Gastroenterology,Gansu Provincial Hospital,from June 2022 to December 2023 were enrolled,and 50 healthy individuals,matched for age and sex,who received physical examination at Physical Examination Center during the same period of time were enrolled as control group.Related data were recorded for all patients,including age,sex,complications,and the results of routine blood test,liver function,and coagulation for the first time after admission.ELISA was used to measure the serum levels of CDK1 and AURKA.A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups;the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and the least significant difference Bonferroni test was used for further comparison between two groups;the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups.The Spearman correlation analysis was used to investigate the correlation between CDK1 and AURKA,and the receiver operating characteristic(ROC)curve and the area under the ROC curve(AUC)were used to investigate the value of CDK1 and AURKA in the diagnosis of HBV-HCC.Results There were significant differences in liver function parameters between the HBV-HCC patients and the control group(all P<0.05);there were significant differences between the CHB group and the HBV-HCC group in albumin,Glb,direct bilirubin,aspartate aminotransferase(AST),gamma-glutamyl transpeptidase(GGT),and alkaline phosphatase(all P<0.05);there were significant differences between the HBV-LC group and the HBV-HCC group in Glb,AST,and GGT(all P<0.05).The HBV-HCC group had significantly higher serum levels of CDK1 and AURKA than the HBV-LC group,the CHB group,and the control group(all P<0.05).There was a significant positive correlation between CDK1 and AURKA in the overall study population and the HBV-HCC patients(r=0.526 6 and 0.815 2,P<0.001).With the control group as reference,CDK1 had an AUC of 0.832 3 in the diagnosis of HBV-HCC,with a sensitivity of 92.86%and a specificity of 75%,and AURKA had an AUC of 0.886 6 in the diagnosis of HCC,with a sensitivity of 95.80%and a specificity of 74%.With the CHB group as reference,CDK1 had an AUC of 0.833 3 in the diagnosis of HBV-HCC,with a sensitivity of 93.75%and a specificity of 75%,and AURKA had an AUC of 0.972 7 in the diagnosis of HBV-HCC,with a sensitivity of 95.83%and a specificity of 91.67%.With the HBV-LC group as reference,CDK1 had an AUC of 0.608 5 in the diagnosis of HBV-HCC,with a sensitivity of 66.67%and a specificity of 54.17%,and AURKA had an AUC of 0.762 2 in the diagnosis of HBV-HCC,with a sensitivity of 95.83%and a specificity of 47.92%.Conclusion The serum levels of CDK1 and AURKA increase with the progression of hepatitis B-associated chronic liver disease,and significant increases in serum CDK1 and AURKA have a certain value in the diagnosis of HBV-HCC.

Objective:To modify the blepharoplasty through palpebral margin incision with preservaton of the pretarsal orbicularis oculi muscle, and to discuss the clinical application effects.Methods:The clinical data of patients who underwent double-eyelid blepharoplasty through palpebral margin incision with preservation of the pretarsal orbicularis oculi muscle were retrospectively analyzed from August 2021 to August 2023 in Changsha Aist Medical Cosmetology Hospital. After the skin peeling rang was designed, the skin was removed, tissue was separated under the orbicularis oculi muscle. The pretarsal orbicularis oculi muscle was thinned-shaped at the upper edge of tarsus as wedge-shaped, and was detached from the upper edge of tarsus. The pretarsal fascia or levator aponeurosis was fixed internally with the orbicularis oculi muscle at the position of the double-eyelid crease. After confirming the shape and symmetry of the double-eyelid, the skin was sutured. Incision healing, eyelid swelling and complications were observed, and double-eyelid morphology was followed up. Six months after surgery, the patients’ satisfaction with the surgical effect (satisfied, somewhat satisfied, dissatisfied) was investigated, and the upper eyelid scar status (including pigmentation, thickness, vascularity, and pliability) was scored using the Vancouver scar scale (VSS). The higher the score, the more serious the scar was.Results:A total of 78 patients with single eyelid or subtle double eyelid were included, including 6 males and 72 females, aged 18-40 years old. All the patients underwent modified double-eyelid blepharoplasty through palpebral margin incision with preservation of the pretarsal orbicularis oculi muscle, 29 patients underwent ptosis correction simultaneously, and 47 patients underwent epicanthus correction simultaneously. The incision healed by first intention after the operation. 78 cases were followed up for 1 to 6 months, and slight upper eyelid swelling was eliminated in 71 cases at 10-15 days and in 7 cases at 25-30 days. Among them, 61 cases were followed up for 6 months, the creases of double-eyelid was natural and smooth, bilateral basic symmetry, and there were no adverse manifestations such as double-eyelid shallowing and pretarsal tissue pilling, and no complications such as incompleted eye-closure. 57 cases were satisfied with the results and 4 cases were somewhat satisfied. The incision scar was not obvious when the eyes were closed in 61 patients. The VSS score was close to 0, and the average score of pigmentation, thickness, vascularity, and pliability were 0.29, 0, 0.31 and 0 points, respectively.Conclusion:This operation method preserves almost all the pretarsal orbicularis oculi muscle to promote the healling of lymph circulation. After the orbicularis oculi muscle is choosed for internal fixation, this more simple operation method can not only guarantee the efficacy of double-eyelid blepharoplasty through palpebral margin incision, but also accelerate the healling of the eyelid after surgery and reduce the incidence of incompleted eye-closure.

Objective:To modify the blepharoplasty through palpebral margin incision with preservaton of the pretarsal orbicularis oculi muscle, and to discuss the clinical application effects.Methods:The clinical data of patients who underwent double-eyelid blepharoplasty through palpebral margin incision with preservation of the pretarsal orbicularis oculi muscle were retrospectively analyzed from August 2021 to August 2023 in Changsha Aist Medical Cosmetology Hospital. After the skin peeling rang was designed, the skin was removed, tissue was separated under the orbicularis oculi muscle. The pretarsal orbicularis oculi muscle was thinned-shaped at the upper edge of tarsus as wedge-shaped, and was detached from the upper edge of tarsus. The pretarsal fascia or levator aponeurosis was fixed internally with the orbicularis oculi muscle at the position of the double-eyelid crease. After confirming the shape and symmetry of the double-eyelid, the skin was sutured. Incision healing, eyelid swelling and complications were observed, and double-eyelid morphology was followed up. Six months after surgery, the patients’ satisfaction with the surgical effect (satisfied, somewhat satisfied, dissatisfied) was investigated, and the upper eyelid scar status (including pigmentation, thickness, vascularity, and pliability) was scored using the Vancouver scar scale (VSS). The higher the score, the more serious the scar was.Results:A total of 78 patients with single eyelid or subtle double eyelid were included, including 6 males and 72 females, aged 18-40 years old. All the patients underwent modified double-eyelid blepharoplasty through palpebral margin incision with preservation of the pretarsal orbicularis oculi muscle, 29 patients underwent ptosis correction simultaneously, and 47 patients underwent epicanthus correction simultaneously. The incision healed by first intention after the operation. 78 cases were followed up for 1 to 6 months, and slight upper eyelid swelling was eliminated in 71 cases at 10-15 days and in 7 cases at 25-30 days. Among them, 61 cases were followed up for 6 months, the creases of double-eyelid was natural and smooth, bilateral basic symmetry, and there were no adverse manifestations such as double-eyelid shallowing and pretarsal tissue pilling, and no complications such as incompleted eye-closure. 57 cases were satisfied with the results and 4 cases were somewhat satisfied. The incision scar was not obvious when the eyes were closed in 61 patients. The VSS score was close to 0, and the average score of pigmentation, thickness, vascularity, and pliability were 0.29, 0, 0.31 and 0 points, respectively.Conclusion:This operation method preserves almost all the pretarsal orbicularis oculi muscle to promote the healling of lymph circulation. After the orbicularis oculi muscle is choosed for internal fixation, this more simple operation method can not only guarantee the efficacy of double-eyelid blepharoplasty through palpebral margin incision, but also accelerate the healling of the eyelid after surgery and reduce the incidence of incompleted eye-closure.

Objective To investigate the effects of calcitriol intervention on PI3K/AKT signaling pathway and wound healing in rats with diabetic foot ulcer(DFU).Methods Thirty-two male SD rats were divided into normal control(Con)group,DFU group,calcitriol low dose(L)group and calcitriol high dose(H)group.A circular wound of 5 mm in diameter and deep to the fascia was made on the dorsum of the left foot of rats in each group.HE staining was used to evaluate the histopathological changes of the wounds.Immunohistochemical method was selected to compare the distribution of CD34-positive cells and the expression of p-PI3K and p-AKT in traumatic tissues of each group.ELISA was adopted in the detection of serum IL-6,IFN-γ,TNF-α and IL-7.RT-PCR was used to detect the mRNA expression of PI3K and AKT in each group,and western blot was used to detect the protein expression of PI3K,p-PI3K,AKT,p-AKT and VEGF.Results Compared with Con group,the expressions of IL-6,IFN-γ,TNF-α,IL-7,CD34,PI3K mRNA,AKT mRNA,p-AKT protein,p-PI3K protein,p-PI3K/PI3K,p-AKT/AKT increased,while PI3K protein expression decreased in DFU,L and H groups(P<0.05),VEGF and AKT protein expression decreased in DFV and L groups(P<0.05).Compared with DFU group,the expressions of VEGF,AKT and PI3K protein increased(P<0.05),while the expressions of p-PI3K/PI3K,p-AKT/AKT decreased in L and H groups(P<0.05),IL-6 decreased in L group(P<0.05),and CD34 expression increased in H group(P<0.05),while IL-6,IFN-γ,TNF-α and IL-7,PI3K mRNA,AKT mRNA,p-AKT protein and p-PI3K protein expression decreased(P<0.05).Compared with L group,the expressions of CD34,VEGF,AKT and PI3K protein increased(P<0.05),while IL-6,PI3K mRNA,AKT mRNA,p-AKT protein,p-PI3K protein,p-PI3K/PI3K and p-AKT/AKT decreased in H group(P<0.05).Conclusions Calcitriol intervention may reduce the activity of the PI3K/AKT signaling pathway,inhibit inflammation,promote neovascularization,and facilitate wound healing in rats with DFU.