[Quantitative HER2 mRNA assay in breast cancer with HER2 immunohistochemistry 0].

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.

[Clinical characteristics of Danon disease].

Objective: To review the clinical data of 7 patients with Danon disease and analyze their clinical characteristics. Methods: The medical records of 7 patients with Danon disease, who were hospitalized in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from April 2008 to July 2021, were reviewed and summarized, of which 6 cases were diagnosed as Danon disease by lysosomal-associated membrane protein-2 (LAMP-2) gene mutation detection and 1 case was diagnosed by clinicopathological features. Clinical manifestations, biochemical indexes, electrocardiogram, echocardiography, skeletal muscle and myocardial biopsy and gene detection results were analyzed, and patients received clinical follow-up after discharge. Results: Six patients were male and average age was (15.4±3.5) years and the average follow-up time was (27.7±17.0) months. The main clinical manifestations were myocardial hypertrophy (6/7), decreased myodynamia (2/7) and poor academic performance (3/7). Electrocardiogram features included pre-excitation syndrome (6/7) and left ventricular hypertrophy (7/7). Echocardiography examination evidenced myocardial hypertrophy (6/7), and left ventricular dilatation and systolic dysfunction during the disease course (1/7). The results of skeletal muscle biopsy in 6 patients were consistent with autophagy vacuolar myopathy. Subendocardial myocardial biopsy was performed in 3 patients, and a large amount of glycogen deposition with autophagosome formation was found in cardiomyocytes. LAMP-2 gene was detected in 6 patients, and missense mutations were found in all these patients. During the follow-up period, implantable cardioverter defibrillator implantation was performed in 1 patient because of high atrioventricular block 4 years after diagnosis, and there was no death or hospitalization for cardiovascular events in the other patients. Conclusion: The main clinical manifestations of Danon disease are cardiomyopathy, myopathy and mental retardation. Pre-excitation syndrome is a common electrocardiographic manifestation. Autophagy vacuoles can be seen in skeletal muscle and myocardial pathological biopsies. LAMP-2 gene mutation analysis is helpful in the diagnose of this disease.

[Complicated lymphatic anomaly: a clinicopathological analysis of four cases].

Objective: To investigate the clinicopathological features, clinical manifestations and different diagnosis of patients with complicated lymphatic anomaly. Methods: The clinical and pathologic data of four patients with complicated lymphatic anomaly diagnosed and treated in Peking Union Medical College Hospital from January 2000 to December 2021 were collected and analyzed. Results: One Gorham-Stout disease case and three generalized lymphatic anomaly cases were included in this cohort. Patients' ages ranged from 7 to 32 years. There were three males and one female. The positions of biopsy included three bone biopsy and one bronchus biopsy. Microscopically, all cases showed diffuse enlarged lymphatic channels. At the same time, osteogenesis was obvious in Gorham-Stout disease case. Radiologically, cortical loss was seen in Gorham-Stout disease, and lytic bone confined to the medullary cavity presented in generalized lymphatic anomaly. The three generalized lymphatic anomaly cases also had coagulopathy, and two had effusion. Conclusions: The histologic feature of complicated lymphatic anomaly was diffuse lymphatic malformation, and the diagnosis depends on clinical and pathologic information. The treatment and prognosis of these diseases are different, and therefore it is necessary to understand their clinical and pathologic features and make the correct diagnosis.

[Application of weighted adjustments of dropout rates in sensitivity analysis of medical repeated measurements data and implementation with SAS].

Objective: To explore the application of weighted adjustments of dropout rates in sensitivity analysis of medically repeated measurements data and the implementation with SAS 9.4 software. Methods: By compiling SAS codes, mixed-effects models for repeated measures were used to conduct the covariance analysis of multivariable repeated measurements data. Meanwhile, the overall dropout rate and the dropout rates of each group were used to make weighted adjustments by applying pattern-mixture models, which was considered to be a sensitivity analysis to validate the stability of results. Results: The dropout rates of placebo group, low-dose and high-dose groups were 8.77%, 11.79% and 16.15%, respectively, the differences were significant (P=0.025). The results of mixed-effects models for repeated measures showed the differences of curative effect indicators changes from baselines of between high-dose, low-dose groups and placebo group were significant (P=0.008 and P=0.002). The results of pattern-mixture models considering weighted adjustments of the respective groups' dropout rates were consistent with those of mixed-effects models for repeated measures. Conclusions: The pattern-mixture models considering weighted adjustments of dropout rates can be used in the sensitivity analysis of repeated measurements data. The SAS codes can provide a practical basis for the popularization and application of weighted adjustments of dropout rates in the sensitivity analysis of repeated measurements data.

[Analysis of the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer].

Objective: To evaluate the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer(NSCLC). Methods: A retrospective analysis of the clinical and pathological data of 138 patients with completely resected N1 NSCLC was conducted. There were 79 males and 59 females, aged from 26 to 81 years with an average of (59±10) years. All of them were treated in the Department of Thoracic Surgery Ⅱ of Peking University Cancer Hospital between January 2007 and December 2015. Patients were stratified based on the 8th edition of the American Joint Committee on Cancer (AJCC) N1 classification and the modified pathological N1 classification strategy, respectively. According to modified pathological N1 classification strategy, which was defined based on the anatomical location of intrapulmonary metastatic lymph nodes, N1 nodes were subcategorized into the hilar (stations 10-11, mN1b) (n=36) and peripheral (stations 12-14, mN1a) (n=102) zones. The Kaplan-Meier curves were plotted to compare the relapse risk and survival analysis, disease-free survival (DFS), and overall survival (OS) were compared between the two staging methods through univariate and multivariate analysis to evaluate the effectiveness of the two classifications in stratifying patients with distinct risks of disease relapse and survival. Results: According to the modified N1 classification, the differences in 5-year DFS and OS between the subgroups (mN1a vs mN1b) were statistically significant(59.5% vs 35.7%; 81.2% vs 56.0%; both P<0.05). However, following the 8th edition of the AJCC N1 classification, no significant differences were found in DFS and OS between the subgroups (both P>0.05). Multivariate analysis showed that the modified N1 classification was an independent prognostic factor to DFS (HR=1.814, 95%CI: 1.005-3.275) and OS (HR=3.919, 95%CI: 1.918-8.009) (all P<0.05). However, the 8th edition of the AJCC N1 classification was not an independent prognostic factor to DFS (HR=1.360, 95%CI:0.767-2.412) or OS (HR=1.620, 95%CI:0.839-3.131) (both P>0.05) as revealed by multivariate analysis. Conclusions: The relapse risk and survival could be assessed effectively using the modified pathological N1 classification, which was defined and subcategorized based on the anatomical location of intrapulmonary metastatic lymph nodes for N1 NSCLC patients. The modified pathological N1 classification is superior to the 8th edition of the AJCC classification.

Characteristics and viral suppression among people living with HIV from the National Free Antiretroviral Therapy Programme, 2019.

OBJECTIVES: This study aimed to present the clinical characteristics of and viral suppression in the national antiretroviral therapy (ART) cohort in China. METHODS: A cross-sectional study was conducted among the participants enrolled in China's National Free Antiretroviral Therapy Programme (NFATP) who were on ART at the end of 2019. The data used were from China's NFATP information system. The primary outcome was viral load (VL) suppression. Full virological suppression and low-level viraemia (LLV) were defined as VL < 50 copies/mL and between 51 and 999 copies/mL, respectively. RESULTS: At the end of 2019, 853 429 people living with HIV (PLWH) were on ART. The median age was 44 years [interquartile range (IQR): 33-54]. The majority of PLWH were 25-44 years old (47.2%) and male (73.5%) and reported HIV infection via heterosexual contact (64.5%). The median (IQR) baseline CD4 count was 257 (140-376) cells/µL. Among the 704 375 PLWH who were on ART for at least 12 months, 82.9% had full virological suppression, 6.5% had VL between 50 and 1000 copies/mL (LLV) and 4.3% had VL > 1000 copies/mL. Optimal full virological suppression occurred in participants who acquired HIV through homosexual contact (88.4%). CONCLUSIONS: Although China can be credited for substantial progress in scaling up ART and viral suppression, challenges remain in some key populations, such as injecting drug users, participants with a history of treatment failure and hepatitis C virus co-infected people. To improve the programme output, additional monitoring and intervention for LLV are necessary.

[Survival time and related factors of antiretroviral therapy among HIV/AIDS patients in Liangshan Prefecture, during 2005-2015].

Objective: To analyze the survival time and to explore the releated factors of antiretroviral therapy among HIV/AIDS patients in LiangShan Prefecture, Sichuan Province for reduction of AIDS death rate. Methods: The retrospective research method was used to collect relevant information from the Management Database of Antiviral Treatment from the National AIDS Comprehensive Prevention Information System. The Kaplan-Meier method was used to describe the survival distribution and to analyze the survival time by single factor and the model of Cox proportional riskanalysis was performed to analyze the survival time of HARRT by multi-factors analysis. Results: Total 14 219 adults and young persons aged ≥15 HIV/AIDS patients received antiviral treatment from 2005 to 2015. The average age of all cases was (36.10±9.41) years old and 10 021 were males (70.5%). The main route of infection was intravenous drug use (61.0%, 8 678 cases). At the end of the observation, 10001 cases (70.3%) were still treated, and 1 425 cases (10.0%) died; Cox Regression analysis showed that female (0.67 (0.55-0.81)), route of sexual infection (0.67 (0.56-0.79)), baseline CD4+T lymphocyte count 200-350 (0.41 (0.35-0.47)) and ≥350 (0.28 (0.24-0.34)), was a protective factor in death. At the beginning of treatment, the patient is clinically staging stage Ⅱ (0.70 (0.58-0.84)) and abnormal BMI (1.75 (1.50-2.03)), is a risk factor for death (P<0.05). Conclusion: Early antiviral treatment is of great significance in improving the anti-viral treatment effect of AIDS. Compliance education should be further strengthened so as to enhance their knowledge. And it is feasible to enhance the effect of treatment through nutritional support for prolonging patients survival time and improving the quality of life.

Continued Elevation of Plasma IL-4 and IL-17 Predicts the Progression from VMC to DCM.

OBJECTIVE: To investigate plasma cytokines (interferon gamma, interleukin-4, and interleukin-17) in patients with viral myocarditis (VMC) and evaluate their predictive value in the progression from VMC to dilated cardiomyopathy (DCM). METHODS: A prospective, multicenter, observational study included 536 patients with newly diagnosed VMC admitted in cardiology departments of 24 tertiary super specialised university-affiliated hospitals in the China registry from January 2012 to June 2016. Demographics and clinical characteristics at baseline and after three months were collected, including laboratory blood tests, ECG, echocardiography, and drug treatment in each participating site. The plasma anti-viral antibodies (Abs), anti-heart autoimmune Abs, and cytokines were detected by ELISA. RESULTS: Of the 536 patients, 534 were included for analysis after two patients died in less than a month. The plasma levels of IFN-γ, IL-4, and IL-17 were continually higher in patients with incident DCM than in those without incident DCM at baseline, from the 1st month and the 3rd month; all had a P value of <0.0001. There was a positive correlation between IL-4 and LVEDd (r = 0.30, P < 0.0001) and between IL-17 and LVEDd (r = 0.11, P = 0.02). When all these covariates have entered the model simultaneously, elevated IL-4 and IL-17 were still significantly associated with DCM incidence. The RR (95% CI) of DCM incidence were 1.04 (1.02-1.06) for IL-4 and 5.24 (2.81-9.79) for IL-17. CONCLUSION: The continued elevation of plasma IL-4 and IL-17 in VMC patients were associated with a high incidence of DCM at three months, and these two cytokines were independent predictors for the progression from VMC to DCM.

[Current status of national free antiretroviral therapy in interprovincial migrating people living with HIV/AIDS and influencing factors, China, 2011-2015].

Objective: To understand the current status of national free antiretroviral therapy in interprovincial migrating people living with HIV/AIDS (PLWHA) and influencing factors in China. Methods: Descriptive and trend test analyses were performed to evaluate the historical characteristics and trends of main descriptive indicators on national free antiretroviral therapy for the interprovincial migrating PLWHA by using the data collected from National Comprehensive HIV/AIDS Information System from 2011 to 2015. Logistic regression model was used to explore the main factors that influencing the coverage of national free antiretroviral therapy among the interprovincial migrating PLWHA in China. Results: The proportion of interprovincial migrating PLWHA gradually increased in last 5 years from 7.1% (17 784/250 645) in 2011 to 10.3% (54 596/528 226) in 2015 (Z=51.38, P<0.000 1) in China. The coverage rate of free antiretroviral therapy in interprovincial migrating PLWHA increased from 37.3% (6 641/17 784) in 2011 to 71.0% (38 783/54 596) in 2015, showing a significant rising tendency (Z=96.23, P<0.000 1), but it was slightly lower than that in non-interprovincial migrating PLWHA in 2015 (71.5%, 338 654/473 630). Multivariate logistic regression analysis showed that the PLWHA who were females, aged ≥50 years, of Han ethnic group, married or had spouse, had the educational level of high school or above, infected through homosexual intercourse, with CD(4)(+)T cells counts ≤500 cells/µl at the first visit, identified to be infected with HIV in medical setting, living in urban areas et al, were more likely to receive free antiretroviral therapy. Conclusions: The coverage rate of free antiretroviral therapy varied among the interprovincial migrating PLWHA with different characteristics. It is still necessary to take effective measures to further increase the coverage of free antiretroviral therapy in interprovincial migrating PLWHA and to include the free antiretroviral therapy in interprovincial migrating PLWHA into standardized management system as soon as possible.

[CD(4+) T lymphocyte responses to anti-retroviral therapy, among HIV/AIDS patients aged 18 and over].

Objective: To compare the differences of CD(4) (+) T lymphocyte (CD(4)) counts between patients aged 18 and over, to explore the effect of age on treatment, 36 months after having received the China National Free AIDS Antiretroviral Treatment on HIV/AIDS. Methods: Through the National ART Information Ssystem, we selected those HIV/AIDS patients who initiated the ART 36 months after the ART, between January 1, 2010 and December 31, 2012 in Guangzhou, Liuzhou and Kunming. Patients were divided into age groups as 18-49, 50-59 and 60 or over year olds, at the baseline of treatment. Under different levels of baseline CD(4) counts, we chose the baseline and different time-point of CD(4) counts as dependent variables, applied mixed linear model to analyze the effects of age, viral suppression, gender, baseline CD(4)/CD(8) ratio and initial treatment regimen. Results: A total of 5 331 HIV/AIDS patients were recruited. No differences were found on age group ratios between different levels of baseline CD(4) counts. At the level of baseline CD(4)<200 cells/µl, both the 50-59 and 60 or above years old groups had lower CD(4) counts than the 18-49 year-old group, within 36 months after the initiation of ART. However, at the baseline CD(4) level of 200-350 cells/µl, no significant differences on CD(4) counts between the 50-59 year-old and 18-49 year-old groups were noticed. CD(4) counts seemed lower in the 60 and above year-old group than in the 18-49 year-old group. Conclusion: Age might serve as an influencing factor on CD(4) counts within 36 months after the initiation of ART, suggesting that earlier initiation of ART might be of help to the recovery of immune function in the 50-59 year-old group.

HIV/AIDS prevention: knowledge, attitudes and education practices of secondary school health personnel in 14 cities of China.

This study assessed the preparedness of school health personnel to develop and deliver HIV/AIDS prevention education programmes for young people in China. A survey of 653 personnel working in secondary schools in 14 cities was conducted. More than 90% had basic knowledge of ways in which HIV can be transmitted, but knowledge of ways in which the virus is not transmitted needs improvement. Substantial numbers of teachers were not sure whether there was an effective preventive vaccine (42%) or did not know whether AIDS was a curable illness or not (32%). The great majority approved of AIDS prevention programmes in universities (98%) and secondary schools (91%), although fewer (58%) agreed that the topic was appropriate for primary schools. Currently, most classroom activities focuses on teaching facts about HIV/AIDS transmission, while less than half are taught about HIV/AIDS related discrimination and life skills to reduce peer pressure. Personnel with some prior training on HIV/ AIDS education (53%) had better factual knowledge, more tolerant attitudes and more confidence in teaching about HIV/AIDS than those without training. The majority of teachers indicated a need for more resource books, audiovisual products, expert guidance, school principal support and dissemination of national AIDS prevention education guidelines to schools.

Single vs twice daily G-CSF dose for peripheral blood stem cells harvest in normal donors and children with non-malignant diseases.

The optimal dose and schedule of G-CSF for mobilization of peripheral blood stem cells (PBSC) is not well defined. G-CSF mobilization was performed in a group of healthy donors and paediatric patients for autologous back-up before receiving allogeneic stem cell transplant. Seventeen consecutive subjects who received G-CSF at 5 microg/kg/dose twice daily (group A) were compared with a historical control group of 25 subjects who received a single daily dose of 10 microg/kg/day G-CSF (group B). Double blood volume apheresis for PBSC collection was started on day 5. G-CSF was continued and apheresis repeated until the targeted CD34+ cell dose was achieved. Both groups were comparable for sex, age, body weight and reason for PBSC collection. Over two-thirds of the subjects in both groups were less than 16 years of age. The G-CSF priming and apheresis were well tolerated. When the first day apheresis products were analyzed, group A resulted in significantly higher yield of total nucleated cells (5.91 vs 3.92 x 108/kg, P = 0. 013), mononuclear cells (5.73 vs 3.92 x 108/kg, P = 0.017), CD34+ cells (2.80 vs 1.69 x 106/kg, P = 0.049) and colony-forming units (107 vs 54 x 104/kg, P = 0.010) as compared with group B. We conclude that the two dose schedule is more efficient in mobilizing PBSC in normal donors and children with non-malignant diseases. This approach may reduce the number of aphereses required and thus reduce the transplant cost.

Partial splenic embolization for hypersplenism concomitant with or after arterial embolization of hepatocellular carcinoma in 30 patients.

PURPOSE: To study the value of partial splenic embolization (PSE) for the treatment of hypersplenism in patients undergoing embolization of hepatocellular carcinoma (HCC). METHODS: Transcatheter hepatic arterial embolization (THAE) combined with PSE was performed in 30 patients with HCC complicating liver cirrhosis, portal hypertension, and hypersplenism. Gelfoam sponge was used as the embolic material for PSE and limited to 100-150 pieces. RESULTS: More than 50% of splenic parenchyma was infarcted in 27 patients. Leukopenia and thrombocytopenia were corrected by PSE in 25 of 27 patients with hypersplenism. In 26 patients with esophageal varices, including 5 patients with bleeding, no rebleeding occurred during a 6-17 month follow-up. Hypersplenism was not corrected in 2 of 3 patients whose infarcted splenic parenchyma was less than 50%. No splenic abscesses or other severe complications were observed. Of the 30 patients treated, 19 are still alive after 1 year. CONCLUSIONS: THAE combined with PSE is a safe and effective measure for patients with HCC.

[Synthesis and antimalarial activities of fluorenemethanols].

For the purpose of improving the oral antimalarial activities of the fluorenemethanols (reported by us in previous articles) which were less effective by oral than by subcutaneous administration, 24 alpha-(alkylaminomethyl)-2, 7-dichloro-9-substituted benzylidene-4-fluorenemethanols (III) were synthesized. The results of preliminary screenings demonstrated that five compounds (No. 1-4, 8) exhibited significant antimalarial activities against Plasmodium berghei NK65 strain in mice by oral administration, at dose of 6.25 mg.kg-1.d-1 x 3 with suppressive rate of 100%. Further evaluation of these 5 compounds showed that 4 of them (No. 1-4) were superior to chloroquine in parallel tests, their ED50 and ED90 were 1.0, 1.6; 0.6, 0.9; 0.7, 1.5 and 0.8, 1.6 mg.kg-1.d-1 x 3 respectively, while the ED50 and ED90 of chloroquine were 1.9 and 2.9 mg.kg-1. d-1 x 3 respectively; one compound (No 8) was equal to chloroquine, its ED50 and ED90 were 1.5 and 3.2 mg.kg-1.d-1 x 3 respectively. Further assessment of these 4 compounds are in progress.

Interactions between drug substances and excipients. 1. Fluorescence and HPLC studies of triazolophthalazine derivatives from hydralazine hydrochloride and starch.

The strength of hydralazine hydrochloride (I) 10 mg tablets, containing starch as an excipient, decreases significantly with time. An investigation to determine the basis for the drop in strength showed that starch exposed to the drug exhibited fluorescence at 414 nm. As model compounds IIIa and V, which contain a triazolophthalazinyl moiety, also fluoresce at about 414 nm, it is proposed that the emission is due to a triazolophthalazine derivative (IIIc) resulting from hydralazine hydrochloride and starch. Degradation of IIIc generates s-triazolo[3,4-a]phthalazine (IIIb), and a small amount of IIIb is detected in aged tablets. IIIc in aged tablets can be converted to IIIb by acid-catalyzed hydrolysis. The quantity of IIIb determined by HPLC amounts to most of the "missing" hydralazine hydrochloride. The reaction between hydralazine hydrochloride and starch is believed to cause the unexpected drop in the strength of hydralazine hydrochloride tablets.