Objective:To investigate HER2 mRNA expression in breast cancer with HER2 immunohistochemistry (IHC) 0 and to analyze the feasibility of distinguishing between the tumor with HER2 μltra-low expression and the one without expression of HER2 (no staining by IHC) by HER2 mRNA level preliminarily.Methods:HER2 mRNA was analyzed by reverse transcription digital PCR in 41 cases of formalin-fixed paraffin-embedded surgical tissue samples of invasive breast cancer obtained between January 2020 and March 2023 at Peking Union Medical College Hospital. The cohort included 21 HER2 IHC 1+ and 20 IHC 0 (12 ultra-low and 8 non-expression of HER2). HER2 mRNA expression level was quantitatively evaluated by the FAM (HER2)/VIC (reference gene) ratio.Results:The expression of HER2 mRNA for the cases with 1+, ultra-low, and non-expression of HER2 by IHC was 0.30 to 1.78 (average 0.90, median 0.82), 0.55 to 1.51 (average 0.93, median 0.90) and 0.22 to 0.78 (average 0.41, median 0.36), respectively. For the mean and median HER2 mRNA levels, there was no significant difference between HER2 IHC 1+ and HER2 ultra-low expression diseases ( P=0.757). A remarkable difference in HER2 gene expression was found between the tumors with 1+ and non-expression of HER2 by IHC ( P=0.002). And, HER2 ultra-low cases contained statistically higher levels of HER2 mRNA compared with non-expression of HER2 subgroup by IHC ( P=0.001). Conclusions:Based on HER2 mRNA, HER2 non-expression and HER2 weak expression (including HER2 IHC 1+ and ultra-low) belong to two different types of the tumor and the disease with HER2 IHC 1+ and HER2 ultra-low expression may be the same. It is necessary to further test the performance of HER2 mRNA detection for stratifying the HER2 weak expression subgroup and to determine the threshold.

Objective:To investigate the clinical characteristics, diagnosis and treatment of chronic natural killer cell lymphoproliferative disease (CLPD-NK) associated peripheral neuropathy.Methods:The clinical data of 1 case of CLPD-NK associated peripheral neuropathy who was hospitalized in Beijing Chaoyang Hospital on March 17, 2022 were reported. The clinical characteristics, laboratory examinations, nerve biopsy, treatment and prognosis of CLPD-NK associated peripheral neuropathy were summarized in combination with domestic and foreign literature reports.Results:This patient was a 62-year-old woman with progressive weakness and numbness of limbs. Nerve conduction test revealed peripheral nerves injures of limbs. No abnormality was found in a routine biochemistry test of cerebrospinal fluid. Lymphocyte infiltration was found in the nerve bundle of sural nerve with CD56(+), TIA-1(+). After prednisone combined with cyclophosphamide treatment, her weakness and numbness of limbs were relieved, and the white blood cells count decreased. A total of 10 cases of CLPD-NK associated peripheral neuropathy were summarized, including 5 males and 5 females. The median onset age was 64 years. The peripheral blood white cells count was 5.50×10 9/L-24.66×10 9/L, the lymphocytes count was 1.54×10 9/L-32.00×10 9/L, and cerebrospinal fluid protein increased in 3 cases. Biopsy of sural nerve showed NK cell infiltration in all patients. Immunosuppression was the main treatment, and most patients had a good prognosis. Conclusions:CLPD-NK associated peripheral neuropathy is rare. Peripheral nerve biopsy is the important diagnostic method. Corticosteroids and immunosuppressants are effective for this disease.

The objective of this study is to elucidate the effects of gut microbiome imbalance on impaired cognitive function in schizophrenia patients， and to propose a new approach to improve cognitive impairment in schizophrenia patients. The cognitive impairment of schizophrenic patients is one of the major barriers that hinder patient social reintegration. With the emergence of microbiome-gut-brain axis， many researchers have found that there is a certain relationship between gut microbiome imbalance and impaired cognitive function in schizophrenia patients， so this paper reviews and summarizes such relationship found in previous studies， with a view to informing the exploration of new ways to improve cognitive impairment in schizophrenia patients.

We presented an adolescent with recurrent intracranial hemorrhage and skin lesion. The diagnosis was unclear and the treatment was difficult. Through a multidisciplinary effort type Ⅰ interferon disease was suspected and later, an interferon-stimulated gene was further detected. Considering the high morbidity and fatality rate of recurrent intracranial hemorrhage, tofacitinib and hydroxychloroquine were administered. After treatment, the livedo reticularis was significantly regressed. Unfortunately, the intracranial hemorrhage recurred due to a pre-existing cerebral aneurysm, leading to death of the patient. The diagnosis and treatment of this case highlight the importance of multidisciplinary collaboration in the diagnosis and treatment of difficult and rare diseases.

Objective:To analyze the thalassemia screening and genotyping in Southwest Guizhou Autonomous Prefecture (referred it as Qianxinan Prefecture), this essay provides the theoretical reference for clinical diagnosis of thalassemia and suspicious cases.Methods:The pregnant women, spouses and neonates who were screened for thalassemia gene in Qian Xi Nan People's Hospital from January 2016 to December 2020 were selected as the research subjects, and peripheral blood or umbilical cord blood samples were collected to extract DNA. The gap-polymerase chain reaction (Gap-PCR) and next-generation sequencing (NGS) technology were used to screen thalassemia, and ArcMap 10.8 software was adopted to map the local spatial distribution of thalassemia based on the screening data.Results:A total of 67 185 cases of people from various regions in Qianxinan Prefecture were screened, and 8 202 cases of thalassemia gene carriers were detected, with a total detection rate of 12.21%. Among them, 5 660 cases of α-thalassemia, with a detection rate of 8.42%; 2 132 cases of β-thalassemia, with a detection rate of 3.17%; 410 cases of αβ complex thalassemia, with a detection rate of 0.61%. In the detection of thalassemia genes, 27 genotypes of α-thalassemia were detected, mainly αα/-α 3.7, accounting for 41.13% (2 328/5 660); 33 genotypes of β-thalassemia were detected, mainly β CD17(A>T)/β A, accounting for 44.09% (940/2 132); 55 genotypes of αβ complex thalassemia were detected, and αα/-α 3.7 complexed β CD17(A>T)/β A dominated, accounting for 21.22% (87/410). There were high incidence areas in the spatial distribution of thalassemia, which were Wangmo County and Ceheng County, and the detection rate was 26.76% (1 438/5 374), 24.39% (1 314/5 387), respectively. Conclusions:The detection rate of thalassemia gene in Qianxinan Prefecture is relatively high, mainly αα/-α 3.7 genotype of α-thalassemia. Wangmo County and Ceheng County are high-incidence areas of thalassemia, and screening efforts should be continued.

Objective: Isolated cortical venous thrombosis (ICoVT) has a low incidence and is easily to be misdiagnosed. The clinical characteristics, diagnosis and treatment of three cases of ICoVT were analyzed in order to improve the level of diagnosis and treatment. Methods: The clinical manifestations, examination, imaging and pathological features of three patients with ICoVT admitted to our hospital were retrospectively analyzed. Results: All the three patients had acute onset. The main symptoms were headache, seizures, numbness of one limb and mental disorders. Intracranial pressure was normal. Brain magnetic resonance imaging (MRI) showed abnormal signals located in cortical sulcus and subcortex. Point and linear hypointensity could be seen on T_2* or susceptibility weighted imaging (SWI) -weighted image. Digital subtraction angiography was performed in one case, showing occlusion of cortical veins. Protein S decreased in two patients, two patients with hyperhomocysteinemia and one patient with antiphospholipid antibody syndrome. The clinical and imaging manifestations of three patients were spontaneously alleviated. Brain biopsy was performed in two patients, and meningeal biopsy was performed in one patient due to significant meningeal thickening. Conclusions The clinical manifestations and routine MRI of ICoVT lack specificity and are easy to be misdiagnosed. However, ICoVT patients are often associated with risk factors for thrombophilia. T_2* or SWI sequences are sensitive to the diagnosis of ICoVT. Brain biopsy has implications for the diagnosis and it is helpful for differential diagnosis, but the indications should be strictly controlled.

Objective To investigate the significance of superb microvascular imaging (SMI) for the detection of neovascularization in carotid atherosclerotic plaques.Methods SMI detected 28 carotid atherosclerotic plaques in 28 patients who underwent carotid endarterectomy.The SMI grade of carotid plaque neovascularization was assigned based on the following grading system:Grade 0,no neovascularization;grade 1,some neovascularization;grade 2,neovascularization observed in the shoulder of the plaque;and grade 3,widely distributed new blood vessels.SMI was used to evaluate the degree of blood flow,maximum flow velocity,and resistance index of the new vessels in the plaque.Microvascular density was measured with CD31 staining.Results Clear correlations were observed between neovascularization density on histopathological staining of the plaque and the degree of blood flow detected with SMI (rs=0.788,P ＜ 0.001).The classification of neovascular blood flow in the plaque was positively correlated with plaque thickness (rs=0.686,P=0.002).Of the 28 cases,neovascularization was detected in 23;of these 23 cases,the direction of blood flow was from the adventitia to the intima in 17 (75％) cases,while 6 (25％) cases showed an irregular direction of blood flow.The mean neovascular flow velocity was (5.620±0.131) cm/s,and the mean neovascular resistance index was 0.660 ± 0.090.Conclusion SMI is useful for the evaluation of carotid atherosclerotic plaque neovascularization.

Objective Isolated cortical venous thrombosis (ICoVT) has a low incidence and is easily to be misdiagnosed. The clinical characteristics, diagnosis and treatment of three cases of ICoVT were analyzed in order to improve the level of diagnosis and treatment. Methods The clinical manifestations, examination, imaging and pathological features of three patients with ICoVT admitted to our hospital were retrospectively analyzed. Results All the three patients had acute onset. The main symptoms were headache, seizures, numbness of one limb and mental disorders. Intracranial pressure was normal. Brain magnetic resonance imaging (MRI) showed abnormal signals located in cortical sulcus and subcortex. Point and linear hypointensity could be seen on T2* or susceptibility weighted imaging (SWI)?weighted image. Digital subtraction angiography was performed in one case, showing occlusion of cortical veins. Protein S decreased in two patients, two patients with hyperhomocysteinemia and one patient with antiphospholipid antibody syndrome. The clinical and imaging manifestations of three patients were spontaneously alleviated. Brain biopsy was performed in two patients, and meningeal biopsy was performed in one patient due to significant meningeal thickening. Conclusions The clinical manifestations and routine MRI of ICoVT lack specificity and are easy to be misdiagnosed. However, ICoVT patients are often associated with risk factors for thrombophilia. T2* or SWI sequences are sensitive to the diagnosis of ICoVT. Brain biopsy has implications for the diagnosis and it is helpful for differential diagnosis, but the indications should be strictly controlled.

Objective To understand the frequency distribution of CYP2C9 and VKORC1 gene single nucleotide polymor-phisms in Yunnan Han population.Methods CYP2C9(430C> T,1075A>C and 1080C> G)locus and VKORC1(-1639G> A and 1173C> T)locus gene polymorphisms in 202 samples were detected by adopting electrochemical gene sensor method,and the allele frequencies and genotype frequencies were performed the statistics and the gene polymorphism in relevant populations was an-alyzed.Results Among 202 samples,202 cases(100.0%)were genotype C/C at CYP2C9 * 2 locus,C allele frequency was 100.0%;185 cases(91.6%)were genotype A/A at CYP2C9*3 locus,15 cases(7.4%)were A/C genotype,2 cases(1.0%)were C/C genotype,A allele frequency was 95.3%,C allele frequency was 4.7%;CYP2C9*5 locus genotype C/C was in 202 cases (100.0%),C allele frequency was 100%;VKORC1 -1639G > A locus genotype A/A was in 145 cases(71.8%),57 cases (28.2%)were G/A genotype,A allele frequency was 85.9%,G was 14.1%;1173C> T locus genotype T/T was in 145 cases (71.8%),C/T gene type in 57 cases(28.2%),T allele frequency was 85.9%,and C was 14.1%.Conclusion The distribution of CYP2C9 gene in Yunnan Han population is similar to that in other regions of our country.The VKORC1 gene is different from the foreign population,Chinese Han nationality and partial minority nationalities.

Objective To evaluate the expression profile of succinate dehydrogenase (SDH)B and SDHC in pheochromocytoma (PCC) and paraganglioma(PGL) (collectively abbreviated as PPGL), and their value in the early diagnosis of malignancy. Methods SDHB and SDHC immunohistochemistry were performed on 140 tumor specimens from 126 PPGL patients (PCC n＝62, PGL n＝61, PCC+PGL n＝3). Results (1) Germline mutation status of 67 patients were determined, of which, identifying 37(55.2%) patients with germline mutation: 2 (3.0%) SDHA, 18 ( 26. 9%) SDHB, 2 ( 3. 0%) SDHC, 5 ( 7. 5%) SDHD, 2 ( 3. 0%) VHL, 7 ( 10. 4%) RET, and 1(1.5%) NF1; and 30 (44.8%) individuals without known mutation. (2) Among 30 PPGLs from 27 patients with SDH-related (SDHx) mutations, 96.7%(29/30) stained negative for SDHB, 76.7%(23/30) stained negative for SDHC, while only 28.6%(14/49) and 18.4%(9/49) stained negative for SDHB and SDHC respectively in the 49 PPGLs without SDHx mutation (P<0.05). (3) The sensitivity of the SDH immunostaining in detecting the presence of germline SDHx mutation was 96.7%for SDHB and 76.7%for SDHC, while the specificity was 71.4%for SDHB and 81.6% for SDHC. ( 4 ) Among PPGLs without SDHB expression, 22. 9% were malignant. This percentage is significantly higher than that in PPGLs with preserved SDHB expression (3.8%, P<0.05). Conclusion SDHB and SDHC immunohistochemistry may serve as post-surgical screening tools to predict the presence of germline SDHx mutation in PPGLs. Negative SDHB expression calls for intense follow-up to rule out malignancy.