Objective: To develop a RHCE genotyping assay based on kompetitive allele-specific PCR (KASP) and assess its clinical accuracy for RhCE blood group determination. Methods: KASP primers were designed to interrogate three RHCE loci: the 109 bp insertion/deletion in intron 2, c. 307T>C, and c. 676C>G. A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology. Discordant samples (n=10) were retested by both methods and further resolved by Sanger sequencing. An additional 377 cases were tested for the c. 48C>G locus to evaluate the predictive accuracy of individual loci and combined locus testing for RhC antigen. Results: Genotyping concordance with serology was 100.0% for both the c. 676C>G locus (RhE/Rhe) and the c. 307T>C locus (Rhc). For RhC prediction using the 109 bp insertion, overall accuracy was 99.7% (1 191/1 194); the 3 discordant cases were confirmed by Sanger sequencing to be false negatives attributable to 109 bp deletion in intron 2. Testing the c. 48C>G allele for RhC prediction yielded 7 false positives, with an accuracy of 98.1% (370/377). RhC antigen status was determined by combining the 109 bp insertion and the c. 48C allele. After excluding 10 samples with inconsistent results between the two loci, the accuracy reached 100% in the remaining 367 samples. When both loci were applied in combination, accuracy reached 100% in the 367 cases with concordant results. Among the 1 194 patients, CCee (45.8%) and CcEe (31.7%) were the most common RhCE phenotypes. The e antigen had the highest positivity rate (92.2%), and the Ce haplotype was the most frequent (66.9%). Conclusion: The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing. Combining the 109 bp insertion/deletion with the c. 48C allele significantly improves RhC antigen prediction compared with either locus alone. This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu, providing local RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.

OBJECTIVE To explore the prevalence of Ureaplasma urealyticum and Mycoplasma hominis among the patients with chronic cervicitis(CC)and observe their drug resistance to commonly used antibiotics.METHODS A total of 91 patients with CC who were treated in gynecology department of Women and Children's Hospital Affiliated to Ningbo University from Jan.2022 to Jun.2024 were assigned as the CC group,meanwhile,91 healthy women who received physical examination were chosen as the control group.The genital tract secretions were collected from all of the research subjects for the culture of U.urealyticum and M.hominis and drug suscep-tibility testing.The isolation rates of U.urealyticum,M.hominis and U.urealyticum plus M.hominis were com-pared between the two groups.The isolation rates of U.urealyticum and M.hominis were compared among the different age groups of CC patients.The drug susceptibility testing of U.urealyticu m and M.hominis for doxycyc-line(DOX),josamycin(JOS),ofloxacin(OFL),clarithromycin(CLA),erythromycin(ERY),tetracycline(TET),azithromycin(AZI)and pristinamycin(PTN)were observed.RESULTS Totally 75(82.41％)genital tract secretion samples tested positive for Mycoplasma among the 91 samples,37 detected with U.urealyticum,25 were M.hominis,and 13 were U.urealyticum plus M.hominis.The isolation rates of U.urealyticum,M.hominis and U.urealyticum plus M.hominis of the CC group were 40.66％,24.47％and 14.29％,respective-ly,higher than 8.79％,4.40％and 5.49％of the control group(P＜0.05).The total detection rate of U.urealyti-cum,M.hominis and U.urealyticum plus M.hominis was higher among the CC patients aged between 20 and 40 years old than among the CC patients aged more than 40 years old(P＜0.05).The U.urealyticum strains from the positive specimens of the CC patients were highly sensitive to CL A and DOS but were resistant to OFL,CIP and PTN;the M.honinis and U.urealyticum plus M.hominis strains were sensitive to JOS and DOX but were resistant to OFL and CIP.CONCLUSIONS The detection rates of U.urealyticum plus M.hominis are higher a-mong the CC patients than among the normal population.The isolated U.urealyticum and M.hominis strains are highly resistant to quinolones and aminoglycosides.It is necessary for the hospital to empirically choose sensitive antibiotics based on the result of drug susceptibility testing.

Kidney transplantation is the best renal replacement therapy for patients with end-stage renal disease. However, it faces significant challenges due to a critical shortage of donor organs and the underutilization of expanded standard donor (ESD) kidneys.Dual kidney transplantation can increase the utilization of expanded standard donor kidneys and enlarge the donor pool, which is an effective solution to deal with kidney shortage. This review provides a systematic presentation of the current status of research on allocation and recipient selection, surgical technique, complications, postoperative efficacy and immunosuppression protocols for adult dual kidney transplantation, with the aim of providing assistance in clinical practice.

OBJECTIVE To explore the prevalence of Ureaplasma urealyticum and Mycoplasma hominis among the patients with chronic cervicitis(CC)and observe their drug resistance to commonly used antibiotics.METHODS A total of 91 patients with CC who were treated in gynecology department of Women and Children's Hospital Affiliated to Ningbo University from Jan.2022 to Jun.2024 were assigned as the CC group,meanwhile,91 healthy women who received physical examination were chosen as the control group.The genital tract secretions were collected from all of the research subjects for the culture of U.urealyticum and M.hominis and drug suscep-tibility testing.The isolation rates of U.urealyticum,M.hominis and U.urealyticum plus M.hominis were com-pared between the two groups.The isolation rates of U.urealyticum and M.hominis were compared among the different age groups of CC patients.The drug susceptibility testing of U.urealyticu m and M.hominis for doxycyc-line(DOX),josamycin(JOS),ofloxacin(OFL),clarithromycin(CLA),erythromycin(ERY),tetracycline(TET),azithromycin(AZI)and pristinamycin(PTN)were observed.RESULTS Totally 75(82.41％)genital tract secretion samples tested positive for Mycoplasma among the 91 samples,37 detected with U.urealyticum,25 were M.hominis,and 13 were U.urealyticum plus M.hominis.The isolation rates of U.urealyticum,M.hominis and U.urealyticum plus M.hominis of the CC group were 40.66％,24.47％and 14.29％,respective-ly,higher than 8.79％,4.40％and 5.49％of the control group(P＜0.05).The total detection rate of U.urealyti-cum,M.hominis and U.urealyticum plus M.hominis was higher among the CC patients aged between 20 and 40 years old than among the CC patients aged more than 40 years old(P＜0.05).The U.urealyticum strains from the positive specimens of the CC patients were highly sensitive to CL A and DOS but were resistant to OFL,CIP and PTN;the M.honinis and U.urealyticum plus M.hominis strains were sensitive to JOS and DOX but were resistant to OFL and CIP.CONCLUSIONS The detection rates of U.urealyticum plus M.hominis are higher a-mong the CC patients than among the normal population.The isolated U.urealyticum and M.hominis strains are highly resistant to quinolones and aminoglycosides.It is necessary for the hospital to empirically choose sensitive antibiotics based on the result of drug susceptibility testing.

Kidney transplantation is the best renal replacement therapy for patients with end-stage renal disease. However, it faces significant challenges due to a critical shortage of donor organs and the underutilization of expanded standard donor (ESD) kidneys.Dual kidney transplantation can increase the utilization of expanded standard donor kidneys and enlarge the donor pool, which is an effective solution to deal with kidney shortage. This review provides a systematic presentation of the current status of research on allocation and recipient selection, surgical technique, complications, postoperative efficacy and immunosuppression protocols for adult dual kidney transplantation, with the aim of providing assistance in clinical practice.

Objective:To investigate the key genetic mutations during the progressive disease (PD) /leukemic transformation (LT) course in MDS by analyzing the dynamic changes of genetic mutations in patients with myelodysplastic neoplasms (MDS) with or without PD/LT.Methods:This study enrolled 84 patients with sequential MDS from May 2019 to August 2023 at ZhongDa Hospital Southeast University and used the next generation sequencing to detect gene mutations. The dynamic changes of genetic mutations in patients with MDS with or without PD/LT were retrospectively analyzed.Results:①This study analyzed data from 84 patients diagnosed with MDS with a median age of 63 (range: 31-95) years and consisting of 51 males and 33 females. Participants were distributed to the PD cohort ( n=20), LT cohort ( n=13), and non-PD/LT cohort ( n=51). Patients from the PD/LT cohorts demonstrated a higher proportion of bone marrow blasts than the non-PD/LT cohort at the first sequencing (1.6% vs. 0.4%, P=0.013). ②The most frequently mutated genes that were detected at first sequencing were ASXL1 ( n=21, 25.0%), TP53 ( n=17, 20.2%), TET2 ( n=12, 14.3%), DNMT3A ( n=11, 13.1%), and U2AF1 ( n=11, 13.1%). Further, patients from the PD/LT cohorts exhibited a higher median number of mutated genes than the non-PD/LT cohort (2 vs.1, P=0.014) at first sequencing. TET2 (27.3% vs. 5.9%, P=0.010), SETBP1 (15.2% vs.2.0%, P=0.033), and RUNX1 (18.2% vs. 2.0%, P=0.013) mutations were enriched in the PD/LT cohorts than in the non-PD/LT cohort. ③The most frequently detected acquired mutations (Ⅰ mutations) and clonally expanded mutations (Ⅱ mutations) were TP53 ( n=9, 10.7%), TET2 ( n=7, 8.3%), ASXL1 ( n=7, 8.3%), and RAS pathway ( n=7, 8.3%). Furthermore, patients from the PD/LT cohorts showed a higher median number of Ⅰ/Ⅱ genes than the non-PD/LT cohort (2 vs. 0, P<0.001), and Ⅰ/Ⅱ RAS pathway (21.2% vs. 0, P=0.001), TP53 (27.3% vs. 0, P<0.001), and TET2 (18.2% vs. 2.0%, P=0.013) mutations were enriched in PD/LT cohorts than in the non-PD/LT cohorts. ④Most of the TP53 mutations (9/12, 75.0%) in PD/LT cohorts were Ⅰ/Ⅱ mutations, whereas all of the TP53 mutations in non-PD/LT cohort were clone-decrease mutations (Ⅲ mutations) (5/8, 62.5%) or clone-stable mutations (Ⅳ mutations) (3/8, 37.5%). Most of the RAS pathway mutations (7/8,87.5%) in the PD/LT cohorts were Ⅰ/Ⅱ mutations, whereas only one patient in the non-PD/LT cohort demonstrated RAS pathway mutations, which belonged to Ⅳ mutations. Conclusion:Patients from the PD/LT cohorts demonstrated a higher proportion of bone marrow blasts and a higher median number of mutations than the non-PD/LT cohort at first sequencing; TET2, SETBP1, and RUNX1 mutations were enriched in the PD/LT cohorts than in the non-PD/LT cohort at first sequencing. Patients from the PD/LT cohorts exhibited a higher number of Ⅰ/Ⅱ mutations than the non-PD/LT cohort. Further, Ⅰ/Ⅱ TP53, RAS pathway, and TET2 mutations were enriched in the PD/LT cohorts, and Ⅰ/Ⅱ TP53 and RAS pathway mutations may contribute to the PD/LT.

Objective:To investigate the nutritional risk status of children in PICU and analyze its influencing factors.Methods:From July 2021 to February 2023,all children aged 1 to 18 years admitted to PICU of Beijing Children's Hospital were investigated by using the pediatric Yorkhill Malnutrition Scoring tool(PYMS)and the clinical data questionnaire.Results:A total of 492 children in PICU were enrolled.The first nutritional risk screening results showed that there were 32 cases of no/low nutritional risk(6.5%),76 cases of medium risk(15.4%),and 384 cases of high risk(78.1%).The incidence of medium/high nutritional risk was as high as 93.5%.The PYMS score of nutritional risk in PICU was(2.61±1.42).The results of multiple linear regression analysis showed that weight,fever time before admission,white blood cells,body mass index,primary diagnosis,father's education,and diet before illness were the main influencing factors of nutritional risk of children in PICU(P<0.05).Conclusion:Children in PICU are in a state of high nutritional risk.It is suggested that children in PICU should carry out nutritional screening in a standardized manner,identify children with high nutritional risk and its influencing factors early.To actively conduct nutritional assessment and nutritional intervention could improve the clinical outcome of children in PICU.

Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

The CyberKnife, an acronym for the stereotactic radiosurgery platform, represents an image-guided stereotactic radiotherapy technique. This technology precisely delivers ionizing radiation to tissues, effectively damaging tumor cells, and is suitable for radiotherapy of both intracranial and extracranial tumors. This article reports the first performance of CyberKnife by radiotherapy at Peking Union Medical College Hospital, including a patient with uncontrolled pituitary adenoma after surgery and radiotherapy, and another patient with vertebral metastasis following targeted therapy for lung adenocarcinoma. The application of CyberKnife technology in radiotherapy has achieved highly accurate dose delivery, enabling targeted irradiation of tumor lesions while minimizing damage to surrounding normal tissues, thereby yielding relatively ideal clinical outcomes.

Objective:To discuss the effect of knock down of gene of kinesin family member 3B(KIF3B),an important component of primary cilia(PC)of in mouse embryonic palatal mesenchymal on the autophagy level of cells(mEPMCs)cells,and to clarify its mechanism.Methods:The mEPMCs from gestational day 14.5 C57BL/6J mice cultured in vitro were collected and divided into control group(administered normal saline),empty lentivirus transfected cell group(sh-NC group)(administered lentivirus transfection),KIF3B knockdown group(sh-KIF3B group)(administered KIF3B gene knockdown),and KIF3B knockdown plus Smoothened receptor agonist(SAG)group(sh-KIF3B+SAG group)(administered KIF3B gene knockdown followed by SAG addition),based on whether the KIF3B gene was knocked down and whether the SAG was used to activate the sonic hedgehog(Shh)signaling pathway and its downstream coreceptor Smo,with 5 rats in each group.Transmission electron microscope was used to observe the morphology and the number of autophagosomes/autolysosomes in the mEPMCs in various groups;Western blotting method was used to detect the expression levels of autophagy-related proteins Beclin-1 and p62,and the Shh signaling pathway proteins Shh and Smo in the mEPMCs in various groups.Results:The transmission electron microscope observation results showed that compared with control group,the number of autophagosomes/autolysosomes in sh-KIF3B group was significantly increased(P<0.05);compared with sh-KIF3B group,the number of autophagosomes/autolysosomes in the mEPMCs in sh-KIF3B+SAG group was significantly decreased(P<0.05).The Western blotting results showed that compared with control group,the Beclin-1 protein expression level in the mEPMCs in sh-KIF3B group was significantly increased(P<0.05),and the KIF3B,p62,Shh,and Smo protein expression levels were significantly decreased(P<0.01);compared with sh-KIF3B group,the Shh,Smo,and p62 protein expression levels in the mEPMCs in sh-KIF3B+SAG group were significantly increased(P<0.01),and the Beclin-1 protein expression level was significantly decreased(P<0.01).Conclusion:Knockdown of KIF3B gene can promote autophagy of the mEPMCs,and the mechanism may be related to its inhibition of the Shh signaling pathway.