Objective:To report a SPTLC2 gene mutation in a family with a phenotype of Charcot-Marie-Tooth disease.Methods:To screen the family of patients with pathogenic mutations of SPTLC2 gene from the database of hereditary peripheral neuropathy in the Department of Neurology, Peking University First Hospital, and to collect their clinical data, peripheral nerve conduction examination, nerve ultrasound examination, pathological examination of the peroneal nerve and whole exome sequencing results of prohand.Results:One family was screened, the proband was a 16-year-old female with 4 years of sensory loss and anhidrosis of both lower limbs and 16 months of walking difficulty who admitted to Peking University First Hospital in January 2022. Physical examination showed sensory loss, dry skin and weakness in distal limbs. Her father had numbness and dry skin in the distal lower limbs from childhood,weakness and atrophy of his lower limbs in adulthood. He died at age of 52 years old. The nerve conduction study revealed no action potentials of the sensory and motor nerves of the lower limbs in the proband. The amplitude of the compound muscle action potential of the motor conduction of the bilateral ulnar nerve and median nerve decreased, and the nerve conduction velocity of the bilateral median nerve were 32 m/s and 24 m/s. Neurosonography showed thickening of peripheral nerves. Sural biopsy revealed severe loss of myelinated and unmyelinated nerve fibers with onion bulbs formation. SPTLC2 gene showed a known heterozygous p.G435V mutation. The lower limb weakness was improved after oral L-serine.Conclusions:SPTLC2 gene mutation can lead to an intermediate Charcot-Marie-Tooth disease phenotype. L-serine can improve the limb weakness.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare hereditary cerebrovascular disease caused by a NOTCH3 mutation. However, the underlying cellular and molecular mechanisms remain unidentified. Here, we generated non-integrative induced pluripotent stem cells (iPSCs) from fibroblasts of a CADASIL patient harboring a heterozygous NOTCH3 mutation (c.3226C>T, p.R1076C). Vascular smooth muscle cells (VSMCs) differentiated from CADASIL-specific iPSCs showed gene expression changes associated with disease phenotypes, including activation of the NOTCH and NF-κB signaling pathway, cytoskeleton disorganization, and excessive cell proliferation. In comparison, these abnormalities were not observed in vascular endothelial cells (VECs) derived from the patient's iPSCs. Importantly, the abnormal upregulation of NF-κB target genes in CADASIL VSMCs was diminished by a NOTCH pathway inhibitor, providing a potential therapeutic strategy for CADASIL. Overall, using this iPSC-based disease model, our study identified clues for studying the pathogenic mechanisms of CADASIL and developing treatment strategies for this disease.

PURPOSE: Sacral nerve stimulation has been used to treat overactive bladder. This study evaluated the effects of stimulation using different pulse widths on the inhibition of bladder overactivity by sacral nerve stimulation (SNM) in pigs. METHODS: Implant-driven stimulators were used to stimulate the S3 spinal nerve in 7 pigs. Cystometry was performed by infusing normal saline (NS) or acetic acid (AA). SNM at pulse widths of 64 μsec to 624 μsec was conducted at the intensity threshold at which observable perianal and/or tail movement was induced. Multiple cystometrograms were performed to determine the effects of different pulse widths on the micturition reflex. RESULTS: AA-induced bladder overactivity reduced the bladder capacity to 46.9%±7.1% of the NS control level (P<0.05). During AA infusion, SNM at 64 μsec, 204 μsec, and 624 μsec increased the bladder capacity to 126.1%±6.9%, 129.5%±7.3%, and 140.1%±7.6% of the AA control level (P<0.05). No significant differences were found among the results obtained using pulse widths of 64 μsec, 204 μsec, and 624 μsec (P>0.05). The actual intensity threshold varied from 0.7 to 8 V. The mean intensity threshold (T visual) for pulse widths of 64 μs, 204 μs, and 624 μs were 5.64±0.76 V, 3.11±0.48 V, and 2.52±0.49 V. T visual for pulse widths of 64 μsec was larger than the other two T visual for pulse widths of 204 μsec and 624 μsec (P<0.05). No significant differences were found among the T visual for pulse widths of 204 μsec and 624 μsec (P>0.05). CONCLUSIONS: This study indicated that different pulse widths could play a role in inhibiting bladder overactivity. It is not yet certain which pulse widths increased bladder capacity compared with AA levels, to minimize energy consumption and maintain patient comfort during stimulation, 204 μsec may be an appropriate pulse width for SNM.
Acetic Acid ; Humans ; Reflex ; Spinal Nerves ; Swine ; Tail ; Urinary Bladder ; Urinary Bladder, Overactive ; Urination

Objective To investigate the antimicrobial resistance profile of the clinical isolates collected from selected hospitals across China. Methods Twenty-nine general hospitals and five children's hospitals were involved in this program. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems. Results were interpreted according to CLSI 2017 breakpoints. Results A total of 190 610 clinical isolates were collected from January to December 2017, of which gram negative organisms accounted for 70.8％ (134 951/190 610) and gram positive cocci 29.2％ (55 649/190 610). The prevalence of methicillin-resistant strains was 35.3％ in S. aureus (MRSA) and 80.3％ in coagulase negative Staphylococcus (MRCNS) on average. MR strains showed much higher resistance rates to most of the other antimicrobial agents than MS strains. However, 91.6％ of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole, while 86.2％ of MRCNS strains were susceptible to rifampin. No staphylococcal strains were found resistant to vancomycin. E. faecalis strains showed much lower resistance rates to most of the drugs tested (except chloramphenicol) than E. faecium. Vancomycin-resistant Enterococcus (VRE) was identified in both E. faecalis and E. faecium. The identified VRE strains were mainly vanA, vanB or vanM type based on phenotype or genotype. The proportion of PSSP or PRSP strains in the non-meningitis S.pneumoniae strains isolated from children decreased but the proportion of PISP strains increased when compared to the data of 2016. Enterobacteriaceae strains were still highly susceptible to carbapenems. Overall, less than 10％ of these strains (excluding Klebsiella spp.) were resistant to carbapenems. The prevalence of imipenem-resistant K. pneumoniae increased from 3.0％ in 2005 to 20.9％ in 2017, and meropenem-resistant K. pneumoniae increased from 2.9％ in 2005 to 24.0％ in 2017, more than 8-fold increase. About 66.7％ and 69.3％ of Acinetobacter (A. baumannii accounts for 91.5％) strains were resistant to imipenem and meropenem, respectively. Compared with the data of year 2016, P. aeruginosa strains showed decreasing resistance rate to carbapenems. Conclusions Bacterial resistance is still on the rise. It is necessary to strengthen hospital infection control and stewardship of antimicrobial agents. The communication between laboratorians and clinicians should be further improved in addition to surveillance of bacterial resistance.

Objective A self-made tiltable treatment couch was adopted for CT simulation positioning and radiotherapy to evaluate the feasibility and effectiveness to minimize the setup errors. Methods Twenty-two patients with thoracic and abdominal tumors receiving radiotherapy in Department of Radiation Oncology,Peking Union Medical College between March and September 2016 were recruited in this study. All patients were randomly divided into the experimental(n=11)and control groups(n=11).In the study group,the tiltable treatment couch was adopted to switch the patients from the standing position to the supine position,and conventional supine position was utilized in the control group. All patients received CT positioning under spontaneous breathing. Image registration was performed according to the standard recommendations of IGRT group. The image registration data for the translational and rotation errors of CBCT were recorded and analyzed. The setup errors were calculated by four-parameter model between two groups. Results In the experimental group,the translational error of the x direction was(-0.012±0.128)cm with a variation range of(0.29-0.70 cm),(0.272±0.123)cm for the y direction(0.23-0.70 cm)and(0.089± 0.105)cm for the z direction(0.14-0.53 cm),respectively. In the control group,the translational error of the x direction was(0.006±0.198)cm(0.27-0.75 cm),(-0.108±0.396)cm for the y direction(0.56-2.08 cm)and(- 0.096± 0.176)cm for the z direction(0.34-0.89 cm), respectively. Conclusions Application of the self-made tiltable treatment couch can enhance the setup reproducibility and reduce the setup errors,especially in the y direction during radiotherapy for the thoracic and abdominal tumors.

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 and FUS mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.
Amyotrophic Lateral Sclerosis ; genetics ; metabolism ; therapy ; Cell Line ; Clustered Regularly Interspaced Short Palindromic Repeats ; Genetic Therapy ; Genome-Wide Association Study ; Humans ; Induced Pluripotent Stem Cells ; metabolism ; Mutation, Missense ; RNA-Binding Protein FUS ; genetics ; metabolism ; Superoxide Dismutase-1 ; genetics ; metabolism

Objective To report the clinical and peripheral neuropathological findings in two patients with autosomal recessive Charcot-Marie-Tooth disease 2K(AR-CMT2K).Methods Case one was a nine year-old girl.She had distal weakness of lower limbs for six years, with calf atrophy and contracture of Achilles tendon for three years.Case two was an eight year-old boy.He had distal weakness of lower limbs with contracture of Achilles tendon and calf muscle atrophy for three years, and proximal weakness of low limbs for two years.The motor nerve conduction velocities in median nerves were 48.1 m/s in case one and 47.6 m/s in case two.The compound motor action potential amplitude of median nerves decreased by 46% in case one and 69% in case two.Sural nerve biopsies and gene targeted next-generation sequencing were performed in both patients.Results Density of myelinated fibers was 8 407/mm2 in case one and 7 714/mm2 in case two.The ratio of myelinated fibers with diameter over 8 μm was 2.6% in case one and 0 in case two.Both patients had small regenerating cluster of myelinated fibers.Thin myelinated fibers appeared in case one.In case two, atypical onion bulb formations with focal folded myelin appeared, and electromicroscopy revealed mitochondrial aggregate in axons.Compound heterozygous mutations of ganglioside-induced differentiation associated protein 1 gene were detected in both patients, including c.767A>G(p.H256R) and c.466G>A (p.A156T) in case one and c.767A>G and 845G>A(p.R282H) in case two.Conclusions Contracture of Achilles tendon may appear in early childhood of AR-CMT2K patients.The main pathological changes in sural nerve are loss of large myelinated fibers, mitochondrial aggregate in axons and myelin abnormalities.

Objective To explore the relationship between the polymorphisms of the primary gout predisposing gene (BCAS3) rs11653176 locus and the incidence of gout in Han Chinese men in coastal areas of Shandong Province. Methods One hundred and fifty-two cases of patients with gout remission,68 cases of acute stage,252 patients with hyperuricemia, and 280 healthy subjects, total males, were enrolled. Genotyping the rs11653176 locus of BCAS3 gene by TaqMan probe technique. The expression level of BCAS3 gene mRNA in each PBMC was measured by RT-qPCR. The levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-18(IL-18)in serum were measured by ELISA. Results The change of allele frequency of rs11653176 locus in BCAS3 gene was associated with gout(P<0.01). BCAS3 mRNA in patients with gout was significantly higher than that of healthy people and patients with hyperuricemia(P<0.01). In gout patients, the expression level of BCAS3 gene containing C allele was higher than that of T allele(mRNA,P<0.05). The inflammatory factors in the acute phase of gout were significantly higher than those in phases of remission and hyperuricemia(P<0. 01). Conclusion Changes in the allele frequency of BCAS3 alleles rs11653176(high C, low T)may contribute to the expression of this gene,and lead to gout. And the onset of gout is closely related to the production of inflammatory factors.

Objective To explore the coincidence rates of positive and reverse blood group of infants younger than six months in Xiamen area in recent years,and to analyze the difference of anti-A or anti-B antibody in infants of different age groups and different ABO blood groups.Methods 2 993 hospitalized infants from 0 to 6 months in five hospitals of Xiamen area were selected.EDTA-K2 anticoagulant blood samples were collected.Micro-column gel agglutination assay was used in ABO blood group.All the infants were divided into six groups,which aged from 0-7 days,8-14 days,15-31 days,1-2 months,3-4 months and 5-6 months.The coincidence rates of positive and reverse blood group of each group were compared.Results Of the 2 993 infants,303 cases were AB type.The coincidence rates of positive and reverse blood group of the other infants which aged from 0-7 days,8-14 days,15-31 days,1-2 months,3-4 months and 5-6 months were 57.0%,61.0%,63.0%,71.8%,84.9% and 91.0%,respectively.The coincidence rates of positive and reverse blood group between different blood groups had no statistically significant difference(x2=0.842,P=0.657).The coincidence rates of positive and reverse blood group had statistically significant difference among different age groups(x2=185.683,P<0.05).Conclusion The IgM antibody of ABO blood group is detected in infants younger than six months.Positive and reverse blood group is necessary in ABO blood group identification of infants to guarantee the safety clinical blood transfusion.

Objective To investigate the susceptibility profile of clinical isolates collected from hospitals across China.Methods Twenty-six general hospitals and four children's hospitals were involved in this program.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were analyzed according to CLSI 2016 breakpoints.Results A total of 153 059 clinical isolates were collected from Junuary to December 2016,of which gram-negative organisms and gram-positive cocci accounted for 71.6％ and 28.4％,respectively.The overall prevalence of methicillin-resistant strains was 38.4％ in S.aureus (MRSA) and 77.6％ in coagulase negative staphylococcus (MRCNS),respectively.The resistance rates of methicillin-resistant strains to most of other antimicrobial agents were much higher than those of methicillin-susceptible strains.However,92.3％ of the MRSA strains were still sensitive to trimethoprim-sulfamethoxazole,while 86.5％ of the MRCNS strains were susceptible to rifampin.No staphylococcal strains were found resistant to vancomycin or teicoplanin.The resistance rates of E.faecalis strains to most drugs tested (except chloramphenicol) were much lower than those of E.faecium.A few strains of both species were resistant to vancomycin.Vancomycin resistant E.faecalis and E.faecium strains were mainly VanA,VanB or VanM type based on their phenotype or genotype.Regarding the non-meningitis S.pneumoniae strains,the prevalence of PSSP or PISP strains isolated from children was higher than that isolated in 2015,but the prevalence of PRSP strains decreased.However,the prevalence of PISP and PRSP strains isolated from adults was lower than that isolated in 2015.The prevalence of ESBLs-producing strains was 45.2％ in E.coli,25.2％ in Klebsiella spp.(K.pneumoniae and K.oxytoca) and 16.5％ in Proteus mirabilis isolates on average.ESBLs-producing Enterobacteriaceae strains were more resistant than non-ESBLs-producing strains in terms of antibiotic resistance rate.The strains of Enterobacteriaceae were still highly susceptible to carbapenems.Overall,less than 10％ of these strains were resistant to carbapenems.About 68.6％ and 71.4％ ofAcinetobacter spp.(A.baumannii accounts for 90.6％) strains were resistant to imipenem and meropenem,respectively.The prevalence of extensively-drug resistant strains in P.aeruginosa was higher than that in 2015.Conclusions Bacterial resistance to commonly used antibiotics is still on the rise.It is necessary to strengthen hospital infection control and management of clinical use of antimicrobial agents,and maintain good practice in surveillance of bacterial resistance.