Objective:To analyze the clinical characteristics and prognosis of primary autoimmune cerebellar ataxia (PACA) patients with autoantibodies.Methods:Patients from the Department of Neurology, Peking Union Medical College Hospital (from March 2013 to December 2023) who met the modified diagnostic criteria of PACA were collected. Cell based assay and tissue based assay were used to detect anti-cerebellar antibodies. The clinical features, results of neuroimaging, cerebrospinal fluid examinations and the prognosis of the patients were analyzed. Modified Rankin Scale (mRS) score≤2 at the last follow-up was defined as a favorable prognosis. Exacerbation of cerebellar ataxia after clinical improvement or stabilization for at least 2 months was defined as relapse.Results:A total of 20 patients were included, including 7 males. The onset age was 48.4 (22.8, 59.3) years. Gait ataxia was the most common cerebellar symptom. Extracerebellar neurological abnormalities included pyramidal sign, peripheral neuropathy/radiculopathy and diplopia. Elevated cerebrospinal fluid white blood cells and positive specific oligoclonal bands were observed in 4/16 and 7/15 of patients, respectively. The brain magnetic resonance imaging examination of the patients showed that 8 patients had no obvious abnormalities, 9 patients showed cerebellar atrophy, and 3 patients showed abnormal signals in the brain or cerebellum. A total of 9 different anti-cerebellar antibodies were detected in the patient′s serum and (or) cerebrospinal fluid, with the most common being anti-Homer-3 antibodies ( n=7). After immunotherapy, 13/17 of patients improved. After 37.5 (21.0, 93.0) months of follow-up, the median mRS score of the patients was 3, and 8 patients (8/20) achieved good prognosis and 6 patients experienced disease recurrence. Conclusions:The clinical manifestations of PACA patients have certain heterogeneity, and positive anti-neuroantibodies and meeting PACA diagnostic criteria are the main basis for diagnosing the disease. Immunotherapy is effective for most patients, but there is still a considerable proportion of patients who have not achieved a good long-term functional prognosis.

Objective:To discuss the effects of kallikrein 5(KLK5)overexpression on the proliferation,invasion and cisplatin(DDP)sensitivity of cervical cancer cells,and to clarify its mechanism.Methods:Western blotting method was used to verify the stable transfection and overexpression of KLK5 in the cervical cancer cell(ME180-OE-KLK5).The cervical cancer ME180-NC-KLK5 and ME180-OE-KLK5 cells in logarithmic growth phase were subcutaneously inoculated into the nude mice to establish the subcutaneous xenograft models.After successful modeling,the mice were randomly divided into normal saline control group(NC-KLK5+0.9％NaCl group),DDP treatment group(NC-KLK5+DDP group),KLK5 overexpression group(OE-KLK5+0.9％NaCl group)and KLK5 overexpression combined with DDP group(OE-KLK5+DDP group),with 5 mice in each group.The nude mice in NC-KLK5+DDP group and OE-KLK5+DDP group were given intraperitoneal injection of DDP at a dose of 5 mng·kg-1;the nude mice in NC-KLK5+0.9％NaCl group and OE-KLK5+0.9％NaCl group were given intraperitoneal injection of normal saline at a dose of 0.01 mL·g-1.The body weights of nude mice were measured every 2 d,and the long diameter and short diameter of the tumors were recorded to calculate the tumor volume and plot the tumor growth curve.At 24 h after the last administration on day 14,the nude mice were sacrificed,and the tumors were dissected and weighed.HE staining method was used to observe the pathomorphology of tumor tissue in the nude mice in various groups;immunohistochemistry staining method was used to observe the expression levels of KLK5,Ki67 and matrix metalloproteinase-9(MMP-9)proteins in the tumor tissues of the nude mice in various groups.Results:Compared with ME180-NC-KLK5 cells,the expression level of KLK5 protein in ME180-OE-KLK5 cells was increased(P＜0.05).In the first week after subcutaneous xenograft inoculation,the nude mice in various groups showed good feeding and activity status,and their body weights gradually increased.The drug administration phase started from the second week.During the drug treatment period,the feeding and activity status as well as body weight of the nude mice in NC-KLK5+0.9％NaCl group showed no significant changes compared with the first week;compared with NC-KLK5+0.9％NaCl group,the nude mice in NC-KLK5+DDP group began to show loss of appetite,no increase in body weight,and decreased activity.During the drug treatment period in the third week,the feeding and activity status of the nude mice in NC-KLK5+0.9％NaCl group showed no significant changes compared with the second week,while they began to show no increase in body weight;compared with NC-KLK5+0.9％NaCl group,the feeding and activity status of the nude mice in NC-KLK5+DDP group were significantly weakened,and their body weights decreased.Compared with NC-KLK5+0.9％NaCl group,the volume of xenograft tumor in NC-KLK5+DDP group was decreased(P＜0.01);compared with NC-KLK5+DDP group,the volume of xenograft tumor OE-KLK5+DDP group was significantly increased(P＜0.001);compared with NC-KLK5+0.9％NaCl group,the volume of xenograft tumor of the nude mice in OE-KLK5+0.9％NaCl group was increased(P＜0.001);compared with OE-KLK5+0.9％NaCl group,the volume of xenograft tumors in the nude mice in OE-KLK5+DDP group showed no statistically significant difference(P＞0.05).Compared with NC-KLK5+0.9％NaCl group,the weight of xenograft tumor of the nude mice in NC-KLK5+DDP group was decreased(P＜0.05);compared with NC-KLK5+DDP group,the weight of xenograft tumors of the nude mice in OE-KLK5+DDP group was significantly increased(P＜0.001);compared with NC-KLK5+0.9％NaCl group,the weight of xenograft tumor of the nude mice in OE-KLK5+0.9％NaCl group was increased(P＜0.001);compared with OE-KLK5+0.9％NaCl group,the weight of xenograft tumors of the nude mice in OE-KLK5+DDP group showed no statistically significant difference(P＞0.05).Compared with NC-KLK5+0.9％NaCl group,the xenograft tumor cells of the nude mice in OE-KLK5+0.9％NaCl group showed greater nuclear heterogeneity;the xenograft tumor cells of the nude mice in OE-KLK5+DDP group and NC-KLK5+DDP group showed cytomorphological changes,manifested as nuclear pyknosis and fragmentation,reduced cell volume,and the appearance of necrosis and apoptosis.Compared with NC-KLK5+DDP group,the degree of necrosis in xenograft tumor of the nude mice in OE-KLK5+DDP group was more pronounced.Compared with NC-KLK5+0.9％NaCl group,the expression levels of KLK5,Ki67 and MMP-9 proteins in xenograft tumor tissue of the nude mice in NC-KLK5+DDP group were decreased(P＜0.05);compared with NC-KLK5+DDP group,the expression levels of KLK5,Ki67,and MMP-9 proteins in xenograft tumor tissue of the nude mice in OE-KLK5+DDP group were increased(P＜0.001);compared with NC-KLK5+0.9％NaCl group,the expression levels of KLK5,Ki67 and MMP-9 proteins in xenograft tumor tissue of the nude mice in OE-KLK5+0.9％NaCl group were increased(P＜0.001);compared with OE-KLK5+0.9％NaCl group,the expression levels of KLK5,Ki67 and MMP-9 in xenograft tumor tissue of the nude mice in OE-KLK5+DDP group showed no statistically significant differences(P＞0.05).Conclusion:KLK5 overexpression can promote the growth of subcutaneous xenograft tumors of cervical cancer ME 180 cells treated with DDP,up-regulate the expressions of Ki67 and MMP-9 in the xenograft tumor tissue,and reduce the sensitivity of the xenograft tumor to DDP.

Objective:To analyze the clinical characteristics and prognosis of primary autoimmune cerebellar ataxia (PACA) patients with autoantibodies.Methods:Patients from the Department of Neurology, Peking Union Medical College Hospital (from March 2013 to December 2023) who met the modified diagnostic criteria of PACA were collected. Cell based assay and tissue based assay were used to detect anti-cerebellar antibodies. The clinical features, results of neuroimaging, cerebrospinal fluid examinations and the prognosis of the patients were analyzed. Modified Rankin Scale (mRS) score≤2 at the last follow-up was defined as a favorable prognosis. Exacerbation of cerebellar ataxia after clinical improvement or stabilization for at least 2 months was defined as relapse.Results:A total of 20 patients were included, including 7 males. The onset age was 48.4 (22.8, 59.3) years. Gait ataxia was the most common cerebellar symptom. Extracerebellar neurological abnormalities included pyramidal sign, peripheral neuropathy/radiculopathy and diplopia. Elevated cerebrospinal fluid white blood cells and positive specific oligoclonal bands were observed in 4/16 and 7/15 of patients, respectively. The brain magnetic resonance imaging examination of the patients showed that 8 patients had no obvious abnormalities, 9 patients showed cerebellar atrophy, and 3 patients showed abnormal signals in the brain or cerebellum. A total of 9 different anti-cerebellar antibodies were detected in the patient′s serum and (or) cerebrospinal fluid, with the most common being anti-Homer-3 antibodies ( n=7). After immunotherapy, 13/17 of patients improved. After 37.5 (21.0, 93.0) months of follow-up, the median mRS score of the patients was 3, and 8 patients (8/20) achieved good prognosis and 6 patients experienced disease recurrence. Conclusions:The clinical manifestations of PACA patients have certain heterogeneity, and positive anti-neuroantibodies and meeting PACA diagnostic criteria are the main basis for diagnosing the disease. Immunotherapy is effective for most patients, but there is still a considerable proportion of patients who have not achieved a good long-term functional prognosis.

Objective:To analyze the clinical characteristics and prognosis of patients with stiff-person syndrome (SPS) associated with glutamic acid decarboxylase (GAD) antibodies.Methods:A retrospective analysis was conducted on demographic characteristics, clinical manifestations, auxiliary examination results, treatment, and prognosis of patients with GAD antibody-related SPS treated at Peking Union Medical College Hospital from January 2015 to July 2023.Results:A total of 33 patients were included, comprising 26 females (78.8%) and 7 males (21.2%), with an onset age of (42±12) years and a disease duration of 24.0 (10.5, 37.5) months. Two cases (6.1%) were diagnosed with tumors, including 1 case with invasive thymoma and 1 case with small cell lung cancer. The majority of patients (87.9%, 29/33) presented with stiffness of trunk and proximal limb muscles, 42.4% (14/33) of patients exhibited episodic spasm, and 54.5% (18/33) of patients were triggered by stimuli such as sound and light. Babinski or Chaddock reflexes were elicited in 33.3% (11/33) of patients. Some patients (36.4%, 16/33) had concurrent limbic encephalitis/epilepsy or cerebellar ataxia (referred to as complex SPS). The median cerebrospinal fluid (CSF) white blood cell count was 2×10 6/L [quartile: 1×10 6/L, 6×10 6/L; range: (0-30)×10 6/L], with mild elevation in 28.0% (7/25) of patients. Multi-channel surface electromyography in 14 out of 21 cases (66.7%) suggested synchronous contraction of agonist and antagonist muscles in a relaxed state. The modified Rankin Scale (mRS) score during the acute phase was 4 (3, 4). All patients received treatment with benzodiazepines or baclofen. Thirty patients (90.9%, 30/33) received first-line immunotherapy, 3 patients (9.1%, 3/33) received second-line immunotherapy with rituximab, and 14 (42.4%, 14/33) received mycophenolate mofetil as long-term immunotherapy. The follow-up period was 16 (10, 42) months, with a median best mRS score of 2; 66.7% (22/33) of patients had a favorable functional prognosis (mRS score≤2), and the recurrence rate was 30.0% (9/30). At the last follow-up, the median mRS score was 2, and 53.3% (16/30) of patients had a favorable functional prognosis. Prognosis was not significantly correlated with gender, age, clinical type, or CSF white blood cell level (all P>0.05). Conclusions:SPS is one of the main clinical phenotypes of GAD antibody-related neuroimmune diseases, commonly observed in middle-aged women, and exhibits a chronic progressive course. Only a minority of patients have concomitant tumors. The diagnosis relies on typical symptoms, GAD antibody testing, and electromyography examination. The initial immune therapy yields good results, but the prognosis for recurrent patients is poor.

Pulmonary hypertension(PH)is a progressive disease characterized by pulmonary vascular remodeling.Current treatments for PH remain suboptimal,and there is an urgent need to better decipher the underlying pathomechanisms to identify new therapeutic targets.MicroRNA(miRNA)are key components in the post-transcriptional machinery that mediates cellular functions and mainly act by regulating the expression of downstream target genes.Numerous in vivo and in vitro studies have demonstrated the involvement of miRNA and their regulators in PH development.However,there is no unified model for the mechanism of miRNA's regulation of pulmonary vascular remodeling.Therefore,this article provides a review on the mechanisms of miRNA in PH characterized in recent years.

Objective:To investigate the clinical and electrophysiological characteristics of facial onset sensory motor neuronopathy (FOSMN) syndrome.Methods:Ten patients diagnosed with FOSMN syndrome in Peking Union Medical College Hospital from January 2012 to December 2022 were included. The clinical and electrophysiological characteristics of patients were analyzed and summarized, and the genetic testing was also performed in these patients.Results:The age of onset was (56.6±6.5) years, and the longest survival duration of disease was 10 years. All patients had numbness around the face and mouth as the first symptom and abnormal blink reflex. A total of 52 sensory nerve conduction nerves were detected, among which 2 median nerves and 2 μlnar nerves showed decreased amplitude of sensory nerve action potential. Needle electromyography showed neurogenic lesions, with both progressive and chronic denervation. Whole exome sequencing identified the heterozygous variant c.272A>C in the exon 4 of the SOD1 gene resulting in the amino acid change p.Asp90Ala in 1 patient. In all patients, the disease progressed relentlessly and eventually led to involvement of respiratory muscle. Conclusion:FOSMN syndrome is characterized by abnormal blink reflex and sometimes abnormal sensory nerve conduction may be shown on electrophysiologic testing.

Objective:To report the clinical characteristics of a case of childhood amyotrophic lateral sclerosis (ALS) caused by SPTLC2 c.778G>A (p.Glu260Lys) mutation. Methods:Whole exon sequencing or whole genome sequencing data from 1 936 patients in the ALS cohort of Peking Union Medical College Hospital were screened for SPTLC2 gene mutations. Clinical data, laboratory examination, neurophysiological examination and genetic test results of the proband were collected. Results:Only one 9-year-old male child with SPLTC2 gene mutation was found. He was admitted to the Department of Neurology, Peking Union Medical College Hospital in December 2022 due to"progressive limb weakness for more than 4 years". Physical examination revealed atrophy and fasciculations of the tongue. Weakness of 4 limbs, muscle atrophy, as well as bilateral hyperreflexia, clonus, and Babinski sign were present. Whole genome sequencing indicated that SPTLC2 gene had c.778G>A (p.Glu260Lys) missense mutation, and no other pathogenic mutations of ALS related genes were detected. Sanger sequencing and family verification showed that neither father nor mother carried the mutation, suggesting that it was a de novo mutation. Nerve conduction velocity test showed no abnormalities, and electromyography suggested neurogenic lesions. Neurofilament light chain in cerebrospinal fluid and serum were increased significantly. The patient′s symptoms continued worsening even after oral administration of L-serine. Conclusion:SPTLC2 gene mutation can cause childhood ALS, and further study of its potential pathogenesis is helpful to uncover another potential pathway of ALS and a novel therapeutic target.

Objective:To explore the effects of mechanical tension on the formation of hypertrophic scars in rabbit ears and transforming growth factor-β 1 (TGF-β 1)/Smad signaling pathway. Methods:The experimental research method was adopted. Six New Zealand white rabbits, male or female, aged 3-5 months were used and 5 full-thickness skin defect wounds were made on the ventral surface of each rabbit ear. The appearance of all rabbit ear wounds was observed on post surgery day (PSD) 0 (immediately), 7, 14, 21, and 28. On PSD 28, the scar formation rate was calculated. Three mature scars in the left ear of each rabbit were included in tension group and the arch was continuously expanded with a spiral expander. Three mature scars in the right ear of each rabbit were included in sham tension group and only the spiral expander was sutured without expansion. There were 18 scars in each group. After mechanical tension treatment (hereinafter referred to as treatment) for 40 days, the color and texture of scar tissue in the two groups were observed. On treatment day 40, the scar elevation index (SEI) was observed and calculated; the histology was observed after hematoxylin eosin staining, and the collagen morphology was observed after Masson staining; mRNA expressions of TGF-β 1, Smad3, collagen Ⅰ, collagen Ⅲ, and α-smooth muscle actin (α-SMA) in scar tissue were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction; and the protein expressions of TGF-β 1, collagen Ⅰ, collagen Ⅲ, and α-SMA, and phosphorylation level of Smad3 in scar tissue were detected by Western blotting. The number of samples of each group in the experiments was 3. Data were statistically analyzed with independent sample t test. Results:On PSD 0, 5 fresh wounds were formed on all the rabbit ears; on PSD 7, the wounds were scabbed; on PSD 14, most of the wounds were epithelialized; on PSD 21, all the wounds were epithelialized; on PSD 28, obvious hypertrophic scars were formed. The scar formation rate was 75% (45/60) on PSD 28. On treatment day 40, the scar tissue of rabbit ears in tension group was more prominent than that in sham tension group, the scar tissue was harder and the color was more ruddy; the SEI of the scar tissue of rabbit ears in tension group (2.02±0.08) was significantly higher than 1.70±0.08 in sham tension group ( t=5.07, P<0.01). On treatment day 40, compared with those in sham tension group, the stratum corneum of scar tissue became thicker, and a large number of new capillaries, inflammatory cells, and fibroblasts were observed in the dermis, and collagen was more disordered, with nodular or swirling distribution in the scar tissue of rabbit ears in tension group. On treatment day 40, the mRNA expressions of TGF-β 1, Smad3, collagen Ⅰ, collagen Ⅲ, and α-SMA in the scar tissue of rabbit ears in tension group were respectively 1.81±0.25, 5.71±0.82, 7.86±0.56, 4.35±0.28, and 5.89±0.47, which were significantly higher than 1.00±0.08, 1.00±0.12, 1.00±0.13, 1.00±0.14, and 1.00±0.14 in sham tension group (with t values of 5.36, 9.82, 20.60, 18.26, and 17.13, respectively, all P<0.01); the protein expressions of TGF-β 1, collagen Ⅰ, collagen Ⅲ, and α-SMA, and phosphorylation level of Smad3 in the scar tissue of rabbit ears in tension group were respectively 0.865±0.050, 0.895±0.042, 0.972±0.027, 1.012±0.057, and 0.968±0.087, which were significantly higher than 0.657±0.050, 0.271±0.029, 0.631±0.027, 0.418±0.023, and 0.511±0.035 in sham tension group (with t values of 5.08, 21.27, 15.55, 16.70, and 8.40, respectively, all P<0.01). Conclusions:Mechanical tension can inhibit the regression of hypertrophic scars in rabbit ears through stimulating the hyperplasia of scars, inhibiting the normal arrangement of dermal collagen fibers, and intensifying the deposition of collagen fibers, and the mechanism may be related to the activation of TGF-β 1/Smad signaling pathway by mechanical tension.

Objective:To observe the effect of pressure therapy on the formation of hypertrophic scars(HTS) and transforming growth factor beta 1 (TGF-β1) / Sma and Mad homolog proteins (Smad) signaling pathway.Methods:Twelve adult healthy New Zealand white rabbits(provided by the Animal Experiment Center of the Air Force Military Medical University) were wounded with 1 cm round punch on 4 sites of the ventral side of each ear. Round scalpels were used to make incisions along the marked lines, dissect the skin and perichondrium. The remaining tissue was scraped off to expose the wound surface. Scar formation was observed on the 28th day after surgery. After the establishment of rabbit ear HTS models, the right ears were used as self-controls, while the left ears were set as the experimental group. Two hypertrophic scars were randomly selected from each rabbit ear, 24 per group. Experimental group: 4-0 nylon silk thread was used to sew the pressure pad on the circular NdFeB magnets pad with a diameter of 1.5 cm to the rabbit ear cartilage. Flexiforce pressure sensor was used to measure the pressure, and the pads were adjusted to maintain a pressure of 20-25 mmHg (1 mmHg=0.133 kPa) for more than 23 h per day. Control group: no treatment. On the 40th day of pressure therapy, the general morphology of rabbit ear scars were observed, and the tissues were harvested for hematoxylin and eosin (HE) staining and Masson’s Trichrome staining for histological study. The scar elevation index (SEI), the number of fibroblasts, and the thickness of the stratum corneum were calculated. The relative mRNA expression levels of TGF-β1, Smad3, collagen type (Collagen )Ⅰ, Collagen Ⅲ, α-smooth muscle actin (α-SMA) were measured with qPCR; Western blotting was used to detect the relative protein expression levels of TGF-β1, Collagen Ⅰ, Collagen Ⅲ, α-SMA and the phosphorylation level of Smad3 (the ratio of p-Smad3 and Smad3 proteins). Statistical analysis was performed with Excel 2019 and GraphPad Prism 8.0. The measurement data conformed to normal distribution and was expressed as Mean±SD. Student’s t-test was used for the comparison between two groups. P<0.05 was considered statistically significant. Results:A total of 96 wounds were formed in 12 rabbits, 27 wounds had no obvious hyperplasia, and the remaining 69 wounds formed hypertrophic scar tissue blocks with a prominent skin surface, firm texture, and dark red appearance. The scars formation rate was 71.9% (69/96). On the 40th day after the application of pressure, the scars in the experimental group were significantly reduced, softer, and the color was slightly lighter compared with the control group. The results of HE staining and Masson’s Trichrome staining showed that the thickness of the stratum corneum, SEI, and the number of fibroblasts were (69.33±6.03) μm, 1.30±0.08, and (236.30±14.64) cells/field, respectively, which were significantly lower than those in the control group [(114.00±10.15) μm, 1.72±0.05, (320.30±14.57) cells/field] (all P<0.01). Abundance in capillaries, inflammatory cells, and fibroblasts were not observed in the dermal layer. The collagen fibers were orderly arranged and sparse. The results of fluorescence quantitative PCR showed that the relative expression levels of TGF-β1, Smad3, Collagen Ⅰ, Collagen Ⅲ, and α-SMA mRNA in the experimental group were 0.48±0.08, 0.58±0.05, 0.04±0.01, 0.15±0.02, 0.31±0.03, respectively, lower than those of the control group(1.00±0.07, 1.00±0.05, 1.00±0.08, 1.00±0.10, 1.00±0.06) (all P<0.01). The results of Western blotting showed that the relative protein expression of TGF-β1, Collagen Ⅰ, Collagen Ⅲ, α-SMA and the phosphorylation level of Smad3 in the experimental group were 0.65±0.03, 0.07±0.01, 0.43±0.03, 0.53±0.03, 0.54±0.03, all lower than the control group’s 1.02±0.06, 0.93±0.05, 0.92±0.03, 0.82±0.03, 0.92±0.03 (all P<0.01). Conclusions:Pressure therapy can significantly inhibit the hyperplasia of scars, improve the structure of HTS tissue, facilitate the normal arrangement of collagen fiber, and reduce the excessive deposition of collagen. Pressure therapy may inhibit scar proliferation by regulating the TGF-β1/Smad signaling pathway.

Paraneoplastic neurological syndromes (PNS) are heterogeneous disorders caused by autoimmune responses of cancer, which can affect any part of the nervous system. Anti-amphiphysin antibody is one of the high-risk PNS antibodies, which is usually associated with small cell lung cancer and breast cancer. However, extrapulmonary neuroendocrine carcinoma is rare in patients with anti-amphiphysin antibody. A case of anti-amphiphysin-associated paraneoplastic brainstem encephalitis with esophageal neuroendocrine carcinoma is reported. The tumor was detected by fluorine 18 fluorodeoxyglucose positron emission tomography and pathologically confirmed by gastroscopic biopsy. The patient′s neurological symptoms were partially improved after treatment of intravenous immunoglobulin and glucocorticoids. However, the disease prognosis is closely related to the accompanying tumor.