[Genotype-phenotype analysis of Fabry disease caused by GLA gene variation in a pedigree].

Objective: To investigate the clinical phenotype and genetic characteristics of patients with Fabry disease caused by a GLA variant, IVS4+919G>A. Methods: It was a prospective study. Fabry disease screening was conducted among high-risk population in Ninghai from October 2021 to August 2023. Those children with decreased α-galactosidase enzyme activity<2.40 µmol/(L·h) or elavated Lyso-GL-3 level>1.10 µg/L in dried blood spot (DBS) method underwent GLA genetic testing for diagnosis confirmation. Meanwhile, family screening was carried out. A proband and his family members diagnosed with Fabry disease were research subjects. The clinical and genetic characteristics of patients with Fabry disease caused by the GLA variant (IVS4+919G>A) were analyzed. Results: The female proband aged 9.8 years with pain in both lower limbs as the initial symptom was found to have a heterozygous GLA variant IVS4+919G>A among 102 patients. In family screening, there were 4 family members (proband's father, elder sister, elder male cousin and elder female cousin) with Fabry disease and a family member (proband's fifth aunt) with a GLA variant. Among these 4 diagnosed family members, the elder male cousin of the proband, a boy aged 13.2 years had a heterozygous GLA variant, IVS4+919G>A with intermittent pain in both lower limbs as the initial symptom. The proband's father had knee joint pain. The proband's elder sister had decreased vision and his elder female cousin had no obvious symptoms. The proband's fifth aunt with a GLA variant had decreased vision. Conclusions: High-risk screening in children and family screening are helpful for early diagnosis and treatment of Fabry disease. Neuropathic pain may be a early symptom in children with Fabry disease caused by the GLA variant, IVS4+919G>A.

Comparison of the Physical Function Trajectories in Three Birth Cohorts of Chinese Older Adults: A 14-Year Longitudinal Study.

OBJECTIVES: It remains uncertain whether the favorable trend of reduction in physical disabilities has become reversed in the recent-born cohorts of older adults. This study aimed to compare the rate of decline with time in self-reported Instrumental Activity of Daily Living (IADL) difficulties, objective measurement of gait speed and grip strength, in three birth cohorts of Chinese older adults. DESIGN: A prospective cohort study. SETTING AND PARTICIPANTS: Four thousand Chinese older adults aged 65 years or above in three birth cohorts (1934-1938, 1929-1933, 1905-1928) were recruited from the community in Hong Kong. MEASUREMENTS: Grip strength, gait speed and IADL difficulties were measured between 2001 to 2017. Joint models were used to examine the trajectories of grip strength, gait speed and IADL difficulties over time, and the interaction effect of age-by-cohort (or also age2-by-cohort) was also examined. RESULTS: The recently born cohort (1934 - 1938) had worse grip strength and more IADL difficulties at the same age than the earlier two cohorts (1929 - 1933; 1905 - 1928). Furthermore, the most recently born cohort also followed a more rapid decline longitudinally with a greater decline observed in gait speed, grip strength and IADL difficulties for women whereas a greater decline in grip strength and IADL difficulties for men. CONCLUSIONS: The continuous improvement of physical limitations in old age may have halted and there appears to be a reversal of this favourable trend in the recent born cohort of older adults living in Hong Kong.

[Expression and significance of immune checkpoint B7-homolog 4 in endometrial cancer].

Objective: To investigate the expression of B7 homolog 4 (B7-H4) and its clinical significance in endometrial cancer. Methods: A total of 833 patients with endometrial cancer admitted to Peking Union Medical College Hospital, Chinese Academy of Medical Sciences from 2009 to 2019, were enrolled. The expression of B7-H4, mismatch repair (MMR), p53, programmed cell death ligand 1 (PD-L1) protein, and CD8+ T lymphocyte density in endometrial cancer tissues were detected by the EnVision two-step method of immunohistochemical staining. First-generation sequencing (Sanger method) was used to determine molecular subtyping of endometrial cancer. The χ2 test was used to compare the differences in positive expression rate of B7-H4 protein in endometrial cancer tissues with different clinicopathological features and molecular subtyping, PD-L1 protein expression, and CD8+ T lymphocyte density. Survival analyses [including recurrence-free survival (RFS) and disease-specific survival (DSS)] were performed for 664 patients with follow-up time≥3 months, with a median follow-up time of 31 months (range: 4-121 months), and the Cox proportional hazards regression model was used to analyze the relevant factors affecting the prognosis of patients with endometrial cancer. Results: (1) The median age of 833 patients was 58 years (range: 25-88 years); pathological type: 595 with endometrioid carcinoma, 238 with non-endometrioid carcinoma; surgical-pathological staging: 542 cases at stage Ⅰ, 38 cases at stage Ⅱ, 173 cases at stage Ⅲ, and 45 cases at stage Ⅳ. Molecular subtyping was performed in 590 patients, including 50 with POLE mutation, 163 with mismatch repair defect (MMR-d) type, 246 with nospecific molecular change (NSMP) type, and 131 with p53 mutation subtype. (2) B7-H4 protein was expressed with brownish-yellow stainind in the cell membrane and cytoplasm of endometrial carcinoma, and the positivity rate of B7-H4 protein was 71.5% (596/833). The positivity rates of B7-H4 protein among patients with different age, surgical-pathological stage, tumor grade, pathological type, depth of muscular invasion, presence or absence of lymphovascular space invasion, and molecular subtype were significantly different (all P<0.05). The positivity rates of B7-H4 protein among patients with different PD-L1 protein expression and CD8+ T lymphocyte density were not significantly different (P>0.05). The 5-year RFS (83.9%) and DSS (87.3%) of B7-H4 protein-positive patients had an increasing trend compared with the 5-year RFS (77.2%) and DSS (78.1%) of B7-H4 protein-negative patients, but there were not statistically significant differences (P=0.053, P=0.083). (3) Univariate analysis showed that the 5-year RFS and DSS of patients with different age, tumor grade, surgical-pathological stage, pathological type, depth of muscular invasion, lymphovascular space invasion, and molecular subtype were significantly different (all P<0.01). There were no significant differences in 5-year RFS (P=0.184, P=0.113) and DSS (P=0.549, P=0.247) among patients with different CD8+ T lymphocyte density and PD-L1 protein expression. Further analysis according to molecular subtype, the results of CD8+ T lymphocyte density and PD-L1 protein expression showed that the 5-year RFS and DSS of B7-H4 protein-positive patients were higher than those of B7-H4 protein-negative patients with NSMP subtype, low density of CD8+ T lymphocyte and PD-L1 protein-negative endometrial carcinoma (all P<0.05), however, there was no significant difference in 5-year DSS between B7-H4 protein-positive patients and B7-H4 protein-negative patients with PD-L1 protein-negative endometrial cancer (P=0.060). Multivariate analysis showed that positive expression of B7-H4 protein was an independent factor for 5-year RFS (HR=0.27, 95%CI: 0.09-0.78, P=0.016) and DSS (HR=0.16, 95%CI: 0.05-0.58, P=0.005) in patients with NSMP subtype endometrial carcinoma. In patients with low-density CD8+ T lymphocytes endometrial cancer, positive expression of B7-H4 protein was an independent factor for 5-year RFS (HR=0.45, 95%CI: 0.26-0.80, P=0.006), but it was not an independent factor for 5-year DSS. In patients with PD-L1 protein-negative endometrial cancer, B7-H4 protein was not an independent factor for 5-year RFS. Conclusion: B7-H4 protein expressed highly in endometrial carcinoma tissues, and its high expression is closely related to clinicopathological features, molecular subtype of p53 mutant and NSMP, and the favorable prognosis of patients with low density of CD8+ T lymphocyte immunophenotype endometrial carcinoma.

[Prognostic factors affecting the success of conversion chemotherapy in patients with unresectable liver metastases from initially colorectal cancer].

Objective: To investigate the factors affecting the success of conversion therapy in patients with initially unresectable colorectal cancer liver metastases (CRLM) in order to provide evidence-based medical evidence for formulating individualized treatment strategies for patients. Methods: A retrospective case-control study was used in this study. Clinical data of 232 patients with initially unresectable CRLM receiving first-line systemic treatment in Sun Yat-sen University Cancer Center from January 2013 to January 2020 were collected, including 98 patients of successful conversion and 134 patients of failed conversion as control. Conversion therapy scheme: 38 patients received FOLFOXIRI regimen chemotherapy (irinotecan, oxaliplatin, calcium folinate and fluorouracil), 152 patients received FOLFOX regimen (oxaliplatin, calcium folinate and fluorouracil), 19 patients received FOLRIRI regimen (irinotecan, calcium folinate and fluorouracil), 23 patients received systemic chemotherapy combined with fluorouridine hepatic artery infusion chemotherapy; 168 patients received targeted therapy, including 68 of bevacizumab and 100 of cetuximab. Logistics analysis was used to compare the factors affecting the success of conversion therapy. The Kaplan-Meier method was used to calculate progression-free survival (PFS), and the Log-rank test was used for survival comparison. Results: Among 232 patients, 98 patients had successful conversions and 134 patients had failed conversions with a successful conversion rate of 42.2%, meanwhile 30 patients underwent simple hepatectomy and 68 underwent hepatectomy combined with intraoperative radiofrequency ablation. After first-line chemotherapy, 111 patients (47.8%) were partial remission, 57 patients (24.6%) were stable disease, and 64 patients (27.6%) were progression disease. During the median follow-up of 18.8 (1.0-87.9) months, 148 patients were dead or with tumor progression. The median PFS time of patients with successful conversion was longer than that of patients with failed conversion (31.0 months vs. 9.9 months, P<0.001). Univariate analysis found that the bilobar distribution of liver tumors (P=0.003), elevated baseline carcinoembryonic antigen (CEA) levels (P=0.024), tumor invasion of the portal vein (P=0.001), number of metastatic tumor>8 (P<0.001), non-FOLFOXIRI (P=0.005), and no targeted therapy (P=0.038) were high risk factors for the failed conversion therapy. The results of multivariate logistics analysis indicated that the number of metastatic tumor >8 (OR=2.422, 95%CI: 1.291-4.544, P=0.006), portal vein invasion (OR=2.727, 95%CI: 1.237-4.170, P=0.008) were the independent risk factors for failed conversion therapy, while FOLFOXIRI regimen (OR=0.300, 95%CI: 0.135-0.666, P=0.003) and targeted drugs (OR=0.411, 95%CI: 0.209-0.809, P=0.010) were independent protective factors for successful conversion therapy. Conclusions: The number of metastatic tumor and portal vein invasion are key factors that affect the outcomes of conversion therapy for initially unresectable CRLM. If a patient can tolerate chemotherapy, a combination program of three-drug and targeted therapy is preferred for the active conversion therapy.

[Clinicopathological analysis of lung metastatic tumor].

Objective: To investigate the clinicopathological features and differential diagnosis of metastatic tumors in the lung. Methods: The clinicopathological data of 226 metastatic tumors in the lung were collected at Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, from January 2014 to December 2018, and the pathomorphological characteristics were analyzed. Results: There were 84 males and 142 females, with an age range from 13 to 77 years. There were 122 patients with multiple pulmonary nodules and 104 patients with solitary pulmonary nodule. The tumors of the highest frequencies were colorectal cancer (n=59), followed by trophoblast tumor (n=44), kidney cancer (n=31), breast cancer (n=20), cervix cancer (n=14), and urinary urothelium cancer (n=8). The time from the diagnosis of primary tumors to metastasis and the status of surgical treatment varied by tumor origin. The morphology of metastatic lung tumors overlapped with that of the primary tumors to some extent. The relative specific morphological characteristics and the presence of carcinoma in situ surrounding the tumors should be carefully searched for to confirm the tumor origin. The metastatic tumors of the lung had morphological characteristics, immunohistochemical TTF1 (-) and tumor of various sources, while the primary tumor differentiation had relatively specific antibodies: colorectal cancer CK20 (+), CDX2 (+), CK7 (-); malignant trophoblastic tumor, HCG (+); renal clear cell carcinoma CD10 (+), vimentin (+), CK7 (-); breast cancer, GATA3 and ER (+); cervical cancer, p16 (+); urothelial carcinoma, CK20, p63 and GATA3 (+). Conclusions: There is overlap between pulmonary metastatic tumor and primary tumor in morphology. Therefore, the diagnosis should be made by combining clinical history, pathological morphology and immunophenotypic characteristics.

[Clinical features and enzyme replacement therapy in 4 children with Fabry disease].

Objective: To analyze the clinical features and efficacy of enzyme replacement therapy in 4 children with Fabry disease. Methods: A retrospective analysis of the clinical manifestations, laboratory findings, genetic variations and treatment were conducted in 4 children with Fabry disease in Children's Hospital of Zhejiang University School of Medicine from January 2014 to July 2020. Results: All four children (2 males, 2 females) with onset age of 12.4 (6.0-16.8) years were diagnosed based on clinical features, α-Gal A enzyme activity, genetic analysis and family history. The clinical manifestations varied in 4 children. All patients had left ventricular hypertrophy and abnormal urinalysis results, 1 case of neuropathic pain, 2 cases of hypohidrosis, 1 case of insipidus, but no angiokeratomas or hearing abnormalities were found. Three missense mutations of GLA gene were identified: c.424T>C (p.C142R), C.335G>A (p.R112H) and c.644A>G (p.N215S). The first two gene mutations were classical phenotypes, and the last one had also been reported in a classic case. In Case 1, no severe adverse events were reported in the first two months of agalsidase beta treatment. The dosage was 1 mg/kg once every 2 weeks. Symptoms of pain intensity and hypohidrosis were improved. Transiently elevated proteinuria was observed but it returned to normal after a week without any treatment. Conclusions: Clinical manifestations of Fabry disease varied in childhood. Multidisciplinary collaboration is required for its early diagnosis and treatment. Severe adverse events are rare in children with short-term therapy of agalsidase beta.

[Epidemiological characteristics of COVID-19 in Wuchang district of Wuhan].

Objective: To analyzes epidemiological characteristics of COVID-19 and provide evidence for adjustment for COVID-19 prevention and control strategies. Methods: The data of COVID-19 cases in Wuchang district reported as of 19 March, 2020 were obtained from National Notifiable Disease Report System of Chinese Disease Prevention and Control Information System. The software's of Excel 2010, SPSSS 22.0, Arc GIS10.2 and Joinpoint regression program 4.8.0.0 were used for statistical analysis. Results: A total of 7547 COVID-19 cases had been reported as of 19 March, 2020 in Wuchang district, including 5 448 confirmed cases (72.19%), 2009 clinical diagnosed cases (26.62%) and 90 asymptomatic cases case (1.19%). The age of the cases was (56.65±16.25) years and age ranged from 2 days to 105 years among confirmed cases, 2634 were males (48.35%) and 2814 were females (51.65%), 2 492 were retirees (45.74%). A total of 545 health workers were infected with SARS-CoV-2 (7.22% of all cases) including 365 confirmed cases and 5 cases have died. A total of 430 cases of death were reported with case fatality rate of 7.89% (430/5 448), case fatality rate of males (10.9%, 266/2 634) was higher than that of females (5.82%, 164/2 814). The first phase of epidemic peak was from January 24 to January 26, the second phase of epidemic peak was from February 1 to February 5 and there was no one of new confirmed case in one day for the first time on March 18. The first four Streets with the highest incidence rates of confirmed cases were Huanghelou Street (1 043.77/100 000), Ziyang Street (627.97/100 000), Yangyuan Street (503.67/100 000) and Shuiguohu Street (486.02/100 000). Compared with females, aged ≤50 years and mild cases of clinical classification respectively, males (RR=0.690, 95%CI: 0.322-1.478), aged >50 years (RR=11.745, 95%CI: 6.878-20.058), severe cases (RR=2.317, 95%CI: 1.789-3.000) and critical cases of clinical classification (RR=10.794, 95%CI: 7.997-14.569), and gender time-dependent covariate (RR=1.392, 95%CI: 1.053-1.840) were major influencing factors of prognosis of COVID-19 confirmed cases. Conclusions: The gender, ages and occupation of distribution were wide among COVID-19 cases in Wuchang district. Males, aged >50 years, severe cases and critical cases of clinical classification were influencing factors of prognosis of COVID-19 confirmed cases. The standardized management of discharged cases, asymptomatic infected cases and close contact persons were main measures to reduce incidence rates of COVID-19 cases.

In Situ Measurement of Vacuum Window Birefringence using 25Mg+ Fluorescence.

Accurate control of the polarization states of laser light is important in precision measurement experiments. In experiments involving the use of a vacuum environment, the stress-induced birefringence effect of the vacuum windows will affect the polarization states of laser light inside the vacuum system, and it is very difficult to measure and optimize the polarization states of the laser light in situ. The purpose of this protocol is to demonstrate how to optimize the polarization states of the laser light based on the fluorescence of ions in the vacuum system, and how to calculate the birefringence of vacuum windows based on azimuthal angles of external wave plates with Mueller matrix. The fluorescence of 25Mg+ ions induced by laser light that is resonant with the transition of |32P3/2,F = 4, mF = 4 â†’ |32S1/2,F = 3, mF = 3 is sensitive to the polarization state of the laser light, and maximum fluorescence will be observed with pure circularly polarized light. A combination of half-wave plate (HWP) and quarter-wave plate (QWP) can achieve arbitrary phase retardation and is used for compensating the birefringence of the vacuum window. In this experiment, the polarization state of the laser light is optimized based on the fluorescence of 25Mg+ ion with a pair of HWP and QWP outside the vacuum chamber. By adjusting the azimuthal angles of the HWP and QWP to obtain maximum ion fluorescence, one can obtain a pure circularly polarized light inside the vacuum chamber. With the information on the azimuthal angles of the external HWP and QWP, the birefringence of the vacuum window can be determined.

[Analytical value of histological research and application of RNAscope technology in liver diseases].

RNAscope is a new generation of in situ hybridization technology and with the advantage of new probe design, it is now being gradually applied to a wide range of research fields, and its research scope is constantly expanding. Our country has large number of liver disease patients, so there is a great demand for histological testing and research evaluation based upon biological information. This technology has unique specificity and sensitivity in situ level, which makes up for the technical defects of immunohistochemistry and traditional in situ hybridization. Realizing the multiplicity and monomolecular level detection, the qualitative and quantitative analysis of nucleic acid level can be carried out in many aspects of the field of liver disease, and the visual evaluation can be achieved by combining it with tissue in situ to highlight the value of "gold standard". This article summarizes the recent year's application of RNAscope technology carried out in liver histology field.

A simple method for in situ measurement of vacuum window birefringence.

We present a simple method to measure the degrees of circular polarization (DoCP) of laser light inside a vacuum chamber and the birefringence of a vacuum window by detecting the fluorescence emitted by Doppler cooled ions in an ion trap. Imperfect laser polarization will cause ions to be pumped to the dark state which will decrease the fluorescence rates of the ions. With a simulation based on the rate equations of the relevant energy levels of 25Mg+ ions, we find that the fluorescence rate is sensitive to the DoCP of the laser. Based on the simulation result, we present a new method to optimize the DoCP of the laser inside the vacuum chamber by adjusting fast axis azimuthal angles of a half-wave plate and a quarter-wave plate outside the vacuum chamber. The laser light is optimized to be circularly polarized with an uncertainty of the DoCP of 7.8 × 10-5. With the obtained polarization information on both sides of the vacuum window and treating the vacuum window as an unknown wave plate, the phase delay and the fast axis azimuthal angle of the vacuum window can be determined in the form of Mueller matrix. The phase delay is determined to be 197.60(39)°, and the fast axis azimuthal angle is determined to be 104.00(5)°.

[Differences of brain functional alterations between subtypes of Cushing's syndrome patients].

Objective: To compare the differences of brain functional damage of subtypes of patients with Cushing's syndrome (CS). Methods: A total of 11 adrenocorticotropic hormone (ACTH)-dependent CS patients and 29 ACTH-independent CS patients were recruited from Chinese PLA General Hospital between September 2015 and March 2017 with confirmed CS. The psychiatric scales and brain task functional magnetic resonance imaging (fMRI) were evaluated. Results: A total of 40 patients (34 females, 6 males) with a mean age of (39.20±12.10) years and a median education level of 12 (9, 16) years were enrolled. ACTH-dependent patients had significantly worse performance than the ACTH-independent patients in response to the depression evaluation (64.6±6.1 vs 56.2±12.8, P=0.008), positive emotion (17.8±4.2 vs 24.3±7.2, P=0.008) and CS life quality [31(29,33) vs 42(29,51), P=0.040]. In the reaction to positive target pictures, ACTH-dependent CS patients showed stronger activation in left superior temporal gyrus compared with patients in ACTH-independent group, while the activation degree of their bilateral dorsal anterior cingulate cortex, bilateralsuperior frontal gyrus and left middle frontal gyrus was much worse. In the reactions to negative target pictures, ACTH-dependent CS patients had weaker activation in bilateral cerebellum, left superior frontal gyrus, left middle frontal gyrus, left precuneus and right postcentral gyrus, compared with patients in the ACTH-independent CS group (P<0.01, AlphaSim corrected). The activation degree of some regions whose brain function was different between the two groups was correlated to the cortisol level, ACTH level, 24 h urinary free cortisol (UFC) level, depression evaluation and negative emotion assessment (all P<0.05). Conclusions: The severity of the depression and the life quality of patients in ACTH-dependent group are worse than ACTH-independent CS patients. The brain function of ACTH-dependent CS patients is much weaker.