Objective To investigate the relationships of the expression levels of BUB1 mitotic checkpoint serine/threonine kinase B(BUB1B)and kinesin family member 11(KIF11)in peripher-al blood with postoperative recurrence in non-muscle-invasive bladder cancer(NMIBC)patients trea-ted with transurethral resection of bladder tumor(TURBT).Methods A retrospective case-control study design was employed.A total of 188 NMIBC patients who received standard TURBT treatment in the 215th Hospital of Shaanxi Nuclear Industry during initial visit from January 2019 to December 2021 were selected as research objects.Patients were followed up for 36 months and divided into re-currence group(n=68)and non-recurrence group(n=120)based on pathological examination re-sults.Additionally,50 healthy individuals who underwent physical examinations during the same period were selected as control group.The levels of BUB1B mRNA and KIF11 mRNA in peripheral blood were compared among the recurrence group,non-recurrence group,and control group.Clinical data were compared between the recurrence and non-recurrence groups.Cox proportional hazards regression models were used to analyze the postoperative follow-up data of NMIBC patients with TURBT.Kaplan-Meier plots were drawn to assess the proportional hazards assumption(PH assumption)of the Cox model.The predictive efficacy of various indicators for postoperative recurrence in NMIBC patients was eval-uated using receiver operating characteristic(ROC)curve.Results The levels of BUB1B mRNA and KIF11 mRNA in peripheral blood in the control group,non-recurrence group,and recurrence group sucessively increased(P＜0.001).The recurrence group had a significantly higher number of patients with pathological T1 staging and multiple tumors compared to the non-recurrence group(P＜0.05).The Kaplan-Meier curves for the number of tumors,pathological staging,BUB1B mRNA,and KIF11 mRNA levels were generally parallel to recurrence rates between the two groups,meeting PH assumption.Multiple tumors,pathological T1 staging,BUB1B mRNA,and KIF11 mRNA levels were independent influencing factors for recurrence in NMIBC patients after TURBT treatment(P＜0.05).ROC curve analysis showed that the areas under the curves(AUCs)for predicting postoper-ative recurrence in NMIBC patients by multiple tumors,pathological T1 staging,BUB1B mRNA,and KIF11 mRNA levels were 0.674,0.746,0.610 and 0.642,respectively.The combined pre-diction using these indicators yielded an AUC of 0.826(95％CI,0.767 to 0.884),with a sensitiv-ity of 86.80％and a specificity of 75.30％,indicating good predictive efficacy.Conclusion Ele-vated levels of BUB1B mRNA and KIF11 mRNA in peripheral blood are significantly associated with postoperative recurrence in NMIBC patients after TURBT and serve as independent risk factors for recurrence.As non-invasive biomarkers,BUB1B mRNA and KIF11 mRNA in peripheral blood com-bined with clinical parameters can significantly improve the predictive efficacy of postoperative recur-rence,providing a new strategy for individualized monitoring.

Objective To investigate the clinical application value of transrectal real-time tissue elastography(TRTE)guided targeted puncture in the diagnosis of prostate cancer.Methods A total of 52 patients with suspected prostate cancer who were treated in the First People's Hospital of Zhengzhou from January 2020 to December 2022 were selected as the study objects.Preoperative routine transrectal ultrasound and TRTE examination were performed to evaluate the benign and malignant prostates.For the 28 patients with suspected lesions found in TRTE,TRTE-guided targeted puncture(2 needles)+ systematic puncture(8 needles)were performed,for the 24 patients with no suspicious lesions found in TRTE,routine ultrasound-guided systematic puncture(12 needles)was performed.The efficacy of TRTE in the diagnosis of prostate cancer was analyzed and the positive rate of targeted puncture and systematic puncture was compared.Results In this study,25 cases of prostate cancer and 27 cases of benign lesions were ultimately pathologically diagnosed,while a total of 28 cases of prostate cancer and 24 cases of benign lesions were diagnosed with TRTE.The positive predictive value and negative predictive value of the diagnosis were 75.0%(21/28)and 83.3%(20/24),respectively.In 28 patients with suspected lesions found by TRTE,a total of 56 needles were targeted puncture,36 needles were diagnosed with prostate cancer,positive rate was 64.29%(36/56),and a total of 224 needles were systematic puncture,89 needles were diagnosed with prostate cancer,with a positive rate of 39.73%(89/224).The positive rate of prostate cancer by targeted puncture was significantly higher than that by systematic puncture(P<0.05).Conclusion TRTE can better diagnose prostate cancer,and its guided targeted puncture has a higher positive rate in the diagnosis of prostate cancer,which can maximize the positive rate of puncture while reducing the number of puncture needles.

Abstract Genetic diagnosis and etiological analysis were performed on a patient with hypoxic-ischemic encephalopathy at birth. MRI technology was used to examine the brain of the child. G-band karyotype analysis technology was used to analyze the karyotype of the child and her parents. Chromosomal microarray analysis(CMA) was used to analyze the entire genome of the child and her parents for chromosomal copy number variation(CNV) and to identify the small supernumerary marker chromosomes. The results of MRI supported the diagnosis of hypoxic-ischemic encephalopathy of the child and found the appearance of Dandy-Walker malformation. Karyotype analysis showed that the mother's karyotype was 46, XX, t(10; 13)(p11. 1; q11)[11]/46, XX[19]. The karyotype of the father was normal. The karyotype of the child was 47, XX, + mar. The CMA results showed that there was no CNVs above 200 kb in the parents. The CMA results of the child showed that the chromosome 10 was repeated in p15. 3 p11. 1, and the fragment size was 38. 39 Mb. In conclusion, this study found a rare small supernumerary marker chromosome(sSMC) on chromosome 10. Its genetic pattern and pathogenicity were analyzed. It is considered that sSMC(10) is the cause of the patient.

【Objective】 To compare the clinical efficacy of percutaneous transforaminal endoscopic decompression （PEID） and percutaneous interlaminar endoscopic decompression （PETD） in the treatment of L5-S1 lateral recess stenosis. 【Methods】 We selected the patients in our center diagnosed with L5-S1 lateral recess stenosis from March 2018 to October 2019 and divided them into Group A and Group B according to the principle of prospective， single-blind， and randomized control （A： PETD； B： PEID）. The operation was performed by the same senior surgeon with mature spinal endoscopy technology. We recorded the basic information， operation duration， usage count of C-arm， hospital stay， VAS score and ODI index of lower back and lower limbs before operation and 3 days， 1 month， 1 year and the last follow-up after the operation， and the operative excellent and good rates （the last follow-up）. The angle of bony lateral recess was measured during pre- and postoperative CT. 【Results】 A total of 95 patients （A： n=48； B： n=47） successfully completed the operation and were followed up for at least 1 year. The two groups did not significantly differ in age， gender， hospital stay， or complication by lumbar intervertebral disc herniation， but PEID group had significantly shorter operation duration and fewer usage counts of C-arm （P<0.001）. VAS score of lower back and lower limbs， and ODI index were significantly reduced at 3 days，1 month， 1 year and the last follow-up after the operation， with no significant difference between the two groups at the same time； no statistical difference was found between the two groups in operative excellent and good rates at the last follow-up （P>0.05）. The postoperative bony side recess angle was significantly improved （P<0.05）， while there was no significant difference in either pre- or postoperative bony side recess angle between the two groups （P>0.05）. 【Conclusion】 Both PEID and PETD are effective strategies in the treatment of L5-S1 lateral recess stenosis and can achieve good clinical outcomes.

Objective:To summarize the clinical phenotype of patients with developmental and epileptic encephalopathy 85 caused by SMC1A gene truncating variation.Methods:The clinical data of 4 patients with epileptic encephalopathy caused by SMC1A gene truncating variation from August 2016 to June 2020 were analyzed retrospectively. Related literatures up to October 2021 with the key words "SMC1A" "Developmental and epileptic encephalopathy 85" "SMC1A, epilepsy" and "SMC1A, truncating" in PubMed, CNKI, and Wanfang databases were searched. Relevant literature was summarized and reviewed.Results:These 4 patients were all female. The onset age of seizure were all in the infantile period. They were admitted to the hospital at 3, 2, 11 and 18 months respectively. Focal seizures occurred in all 4 patients, while 1 of them experienced infantile spasm. The characteristic of cluster was observed in all of them with an interval of 14 days to 5.0 months. The seizures were all refractory to different kinds of anti-seizure medications. All 4 patients had severe developmental retardation with microcephaly (head circumference<-2 s). The interictal electroencephalogram (EEG) was characterized by diffuse slow wave. The 4 SMC1A gene variants were p.Gly655fs, p.Glu811fs, p.Arg412fs and p.Ile143fs, all of which were de novo frameshift variation after parental validation. There were another 17 cases with SMC1A gene truncating variation reported in 6 English articles and 1 Chinese article. Among these 21 patients, who were all female, the onset of seizures occurred between 0.5 and 18.0 months of age. Seventeen cases (81%) had the characteristics of cluster attacks, and the intervals of attack cycles were different. Seizure types included generalized tonic-clonic seizure (12 cases (57%)), focal seizure (11 cases(52%)), myoclonic(4 cases(19%)), spasm (4 cases(19%)), atypical absence (3 cases(14%)), tonic seizure (2 cases (10%)), and atonia (1 case(5%)). In addition, 4 cases (19%) had status epilepsy. All patients had moderate to severe mental retardation. Microcephaly was found in all patients. Among 18 cases,EEG in 8 cases had diffuse slow wave background. Brain magnetic resonance imaging (MRI) was normal in 13 cases (62%). Other MRI changes included cerebellar atrophy (3 cases), thin corpus callosum (3 cases), and lateral ventricular enlargement (2 cases). Twenty patients did not respond well to antiepileptic drugs. Conclusions:The clinical phenotypes of patients with epilepsy encephalopathy 85 caused by SMC1A gene truncating variation are characterized by female, early-onset, clustering of seizures, development delay and microcephaly. Diffuse slow waves are shown in interictal EEG in partial. Response to treatment and prognosis are poor.

The CRISPR/Cas9 gene editing system has been widely used in basic research, gene therapy and genetic engineering due to its high efficiency, fast speed and convenience. Meanwhile, the discovery of novel CRISPR/Cas systems in the microbial community also accelerated the emergence of novel gene editing tools. CRISPR/Cpf1 is the second type (V type) CRISPR system that can edit mammalian genome. Compared with the CRISPR/Cas9, CRISPR/Cpf1 can use 5'T-PAM rich region to increase the genome coverage, and has many advantages, such as sticky end of cleavage site and less homologous recombination repair. Here we constructed three CRISPR/Cpf1 (AsCpf1, FnCpf1 and LbCpf1) expression vectors in silkworm cells. We selected a highly conserved BmHSP60 gene and an ATPase family BmATAD3A gene to design the target gRNA, and constructed gHSP60-266 and gATAD3A-346 knockout vectors. The efficiency for editing the target genes BmATAD3A and BmHSP60 by AsCpf1, FnCpf1 and LbCpf1 were analyzed by T7E1 analysis and T-clone sequencing. Moreover, the effects of target gene knockout by different gene editing systems on the protein translation of BmHSP60 and BmATAD3A were analyzed by Western blotting. We demonstrate the CRISPR/Cpf1 gene editing system developed in this study could effectively edit the silkworm genome, thus providing a novel method for silkworm gene function research, genetic engineering and genetic breeding.
Animals ; Bombyx/metabolism* ; CRISPR-Cas Systems/genetics* ; Endonucleases/genetics* ; Gene Editing ; RNA, Guide/genetics*

【Objective】 To investigate the effect of decompression technique with bone-chisel under percutaneous spinal endoscopy through transforaminal approach in elderly patients with unilateral lateral recess stenosis （ULLRS）. 【Methods】 We enrolled 46 old patients with ULLRS who underwent percutaneous spinal endoscopy through transforaminal approach assisted with bone-chisel technique from March 2017 to July 2019. The visual analogue scale （VAS） score of low back pain and leg pain， the Oswestry dysfunction index （ODI）， and the modified MacNab score before and after operation were obtained. We also recorded preoperative and postoperative sagittal diameter measured by computer tomography （CT） to evaluate decompression of the lateral recess. All the patients were followed up for at least 1 year for complications. 【Results】 All the 46 patients were followed up for 17 to 42 months. Dural tear occurred in one case， transient nerve root irritation in two cases， and there were no other complications. The low back pain VAS score， lower limb pain VAS score， and ODI index of all the patients were significantly improved in the last follow-up （P<0.05）. The results of the modified MacNab efficacy evaluation at the last follow-up revealed that the excellent and good rate was 91.3% （excellent in 38， good in 4， and fair in 4）. The sagittal diameter （mm） of the bony lateral recess 1 year after surgery increased by an average of 55.8%， which was significantly better than that before surgery （t=6.354， 4.22±1.25 vs. 2.71±0.57， P<0.001）. 【Conclusion】 Percutaneous spinal endoscopy through transforaminal foramen approach combined with bone-chisel technique is effective in treating ULLRS in elderly patients.

Objective:To analyze the brain functional fluctuation of benign epilepsy in children with central-temporal spikes(BECTS) by using ReHo algorithm based on the resting-state brain functional imaging, and to explore the connection of the brain function and changes of the connection pattern, so as to find the damage of the cognitive function of BECTS children in the early stage.Method:s Perspectiveness and simple random selection of 20 BECTS children and 20 healthy control children admitted to Shenzhen Children′s Hospital from January 2015 to December 2017 were conducted for basic information collection and functional magnetic resonance imaging (fMRI) testing in a resting-state.Result:s Significantly lower ReHo value appeared in the default mode network (DMN) area, and the precuneus (voxel=422, t=-5.085 6), cuneus (voxel=85, t=-4.240 3), angular gyrus (voxel=191, t=-4.681 2), cingulate cortex (voxel=313, t=-5.238 2), anterior central gyrus (voxel=12, t=-3.482 7), and supplementary motor area (voxel=1 356, t-6.596 2). The significantly increased ReHo was found in the bilateral cerebellum (voxel=71, t=5.658 2), right superior temporal gyrus (voxel=24, t= 5.184 0), operculum insulae (voxel=337, t=6.814 9), left parietal lobe (voxel=12, t=4.378 7), and inferior parietal lobule (voxel=11, t=3.433 7). Conclusions:Significant impairment of DMN, Wernicke and angular gyrus functions in BECTS children may be one of the mechanisms of cognitive dysfunction.Enhanced sensorimotor area and cortical brain functions near the operculum insulae and central sulcus lead to seizures with typical clinical symptoms.fMRI has a high specificity and sensitivity for evaluating the brain function of children with BECTS, and it can detect the impairment of cognitive function in children with this type of epilepsy at an early stage.

Objective: To investigate the efficacy and safety of ketogenic diet (KD) and antiepileptic drugs(AEDs) in the children with drug refractory Dravet syndrome (DS). Methods: Thirty-two cases of drug refractory DS were enrolled into the Department of Neurology, Shenzhen Children′s Hospital Affiliated to Shantou University Medical School from July 2016 to December 2017, and they were divided into 2 groups: KD group and AEDs group (16 cases for each group), respectively.KD was added to as an additional therapy for KD group, and oral AEDs were administered only in AEDs group.In KD group, oral AEDs were not adjusted for the first 3 months.AEDs could be adjusted within a limited range in 2 groups after 3 months.The clinical efficacy, improvement of cognitive function, retention rate and side effects were observed and compared after 3, 6, 12 months of treatment.The average monthly seizure frequency within 3 months before enrollment was recorded as the baseline.The clinical efficacy was assessed by comparing the seizure frequency of each observation period with the baseline. Results: In KD group, after 3, 6, 12 months′ follow-up, KD the-rapy was maintained in 15, 14, 12 patients.The number of patients whose seizure reduction over 50% was 10, 12, 11 cases, respectively.The number of patients whose seizure reduction over 90% was 7, 9, 10 cases, respectively.The number of patients who were seizure free was 3, 6, 8 cases, respectively.In AEDs group, after 3, 6, 12 months′ therapy, the number of patients whose seizure reduction over 50% was 6, 7, 8 cases, respectively, the number of patients whose seizure reduction over 90% was 3, 3, 4 cases, respectively.The number of patients who were seizure-free was 2, 1, 2 cases, respectively.There was a significant difference in the seizure reduction between 2 groups after 6, 12 months (P<0.05). Furthermore, the incidence of status epilepticus (SE) was significantly reduced in KD group, and non-fever related status epilepticus (NFSE) was preferentially improved.There was a significant difference in the incidence of SE between before and after treatment in the KD group (P<0.05). After 12 months, there was a significant difference in the incidence of SE between 2 groups (P<0.05). After 6, 12 months of treatment, the patients in KD group had significant improvements in adaption, gross motor and language quotients by Gesell Developmental Scale compared to the AEDs group (all P<0.05). Eleven of 12 children who adhered to the therapy for 1 year in KD group had improvement of developmental quotient ≥ 1 grade, however, 7 cases of 16 children in AEDs group had this improvement.The incidence of adverse effect in the KD group and the AEDs group was 37.5%(6/16 cases) and 56.3%(9/16 cases), respectively, and the difference was not significant (P>0.05). Conclusions KD can not only reduce seizure frequency and relieve SE, but also improve the cognitive function of drug refractory DS.The adverse reaction ratio of KD does not increase significantly compared to AEDs.Therefore, KD is effective and safe therapy for children with drug-resistant DS.

Objective To observe the effects of Tocilizumab on white blood cell (WBC) after acute myocardial infarction (MI) and explore its potential to treat MI.Methods Rats were divided into 3 groups:control,MI,and MI treated.Serum from individual mouse was collected before and after subcutaneously Tocilizumab treatment.The level of interleukin-6 (IL-6),the number of WBC and the ratio of active hematopoietic stem cell (HSC)was tested by ELISA,flow cytometry and blood routine examination.The fibrosis of heart tissue was observed by immunohistochemistry.Results The IL-6 level and the number of the WBC were reduced after Tocilizumab treatment.It indicates the effect of inhibiting the activity of HSC and improving the situation of cardiocytes remodeling.Conclusion Tocilizumab could inhibit the generation of WBC and re-construct myocardium after MI.