1.Polysaccharide of Alocasia cucullata Exerts Antitumor Effect by Regulating Bcl-2, Caspase-3 and ERK1/2 Expressions during Long-Time Administration.
Qi-Chun ZHOU ; Shi-Lin XIAO ; Ru-Kun LIN ; Chan LI ; Zhi-Jie CHEN ; Yi-Fei CHEN ; Chao-Hua LUO ; Zhi-Xian MO ; Ying-Bo LIN
Chinese journal of integrative medicine 2024;30(1):52-61
OBJECTIVE:
To study the in vitro and in vivo antitumor effects of the polysaccharide of Alocasia cucullata (PAC) and the underlying mechanism.
METHODS:
B16F10 and 4T1 cells were cultured with PAC of 40 µg/mL, and PAC was withdrawn after 40 days of administration. The cell viability was detected by cell counting kit-8. The expression of Bcl-2 and Caspase-3 proteins were detected by Western blot and the expressions of ERK1/2 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). A mouse melanoma model was established to study the effect of PAC during long-time administration. Mice were divided into 3 treatment groups: control group treated with saline water, positive control group (LNT group) treated with lentinan at 100 mg/(kg·d), and PAC group treated with PAC at 120 mg/(kg·d). The pathological changes of tumor tissues were observed by hematoxylin-eosin staining. The apoptosis of tumor tissues was detected by TUNEL staining. Bcl-2 and Caspase-3 protein expressions were detected by immunohistochemistry, and the expressions of ERK1/2, JNK1 and p38 mRNA were detected by qRT-PCR.
RESULTS:
In vitro, no strong inhibitory effects of PAC were found in various tumor cells after 48 or 72 h of administration. Interestingly however, after 40 days of cultivation under PAC, an inhibitory effect on B16F10 cells was found. Correspondingly, the long-time administration of PAC led to downregulation of Bcl-2 protein (P<0.05), up-regulation of Caspase-3 protein (P<0.05) and ERK1 mRNA (P<0.05) in B16F10 cells. The above results were verified by in vivo experiments. In addition, viability of B16F10 cells under long-time administration culture in vitro decreased after drug withdrawal, and similar results were also observed in 4T1 cells.
CONCLUSIONS
Long-time administration of PAC can significantly inhibit viability and promote apoptosis of tumor cells, and had obvious antitumor effect in tumor-bearing mice.
Mice
;
Animals
;
Alocasia/metabolism*
;
MAP Kinase Signaling System
;
Caspase 3/metabolism*
;
Apoptosis
;
RNA, Messenger/metabolism*
2.Pathological characteristics of angioimmunoblastic T cell lymphoma with bone marrow involvement
Huilan LI ; Kun RU ; Xiaoyan LI ; Lidan SUN ; Fengli LI ; Jingya YAO ; Yani LIN ; Enbin LIU
Chinese Journal of Clinical and Experimental Pathology 2024;40(1):51-55
Purpose To explore the pathological features of angioimmunoblastic T-cell lymphoma(AITL)with bone marrow involvement and to improve awareness of bone marrow infiltration in AITL.Methods The tissue morphology of 32 cases of AITL with bone marrow involvement was retrospectively analyzed.Im-munohistochemistry using the EnVision method and ten-color flow cytometry were conducted to detect AITL-related immune markers.T cell clonality was analyzed through T cell receptor(TCR)gene rearrangement.Results The predominant pat-terns of tumor cell infiltration were nodular(20/32,62.5%)and interstitial or small clusters(10/32,31.3%).The nodules showed a mixture of cellular components.In some cases,the fo-ci contained a mixture of cells with characteristic"granuloma-toid"changes.The tumor cells were mainly small to medium-sized lymphocytes with inconspicuous atypia.Some cases showed plasma cell proliferation.19 cases were subject to immunohisto-chemical staining,which revealed a low count of CD4-positive T cells,with an average of 8.4%.The positive rates of T follic-ular helper cells(TFH)markers were as follows:CD10(7/14,50.0%),BCL6(6/19,31.6%),PD-1(13/19,68.4%),and CXCL13(13/19,68.4%).In most cases,tumor cells showed co-expression of PD-1 and CXCL13,but the number of positive cells was less than 1%.Flow cytometry analysis was performed in 24 cases,among which 22 cases all consistently expressed cytoplasmic CD3(cCD3),CD5,CD4,and CD2,with varying degrees of CD10 expression.In some cases,there was a lack of expression of surface CD3(sCD3)(12/22,54.5%),while there was a lack of expression of CD7(8/22,36.4%).and no abnormal T cells were found in 2 cases.TCR gene rearrangement analysis was performed in 7 cases,with 3 cases showing TCR clonality.Conclusion AITL with bone marrow involvement exhibits a lower proportion of tumor cells and less atypia,making it prone to misdiagnosis.The presence of lymphocytic foci with mixed cellular components in the bone marrow can indicate bone marrow involvement in AITL.Flow cy-tometry detection of abnormal T cells(double positive for CD4 and CD10)strongly suggests bone marrow infiltration in AITL.A comprehensive diagnosis of bone marrow involvement in AITL re-quires consideration of bone marrow biopsy,flow cytometry,and TCR gene rearrangement analysis.
3.Comparison of quantitative detection of BCR::ABL1 p210 transcript levels: a multicenter study
Chuting ZHAO ; Canrong NI ; Yani LIN ; Xiaoli MA ; Qisheng WU ; Fang WANG ; Xiaoxue HAN ; Feng LIU ; Yang XU ; Hongxing LIU ; Jie CHEN ; Kun RU ; Minghua ZHU
Chinese Journal of Pathology 2024;53(7):672-677
Objective:To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories.Methods:The interlaboratory comparison was carried out in two stages. The samples were prepared by the reference laboratory. The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%, 0.01%-0.1%, 0.1%-1%, 1%-10% and>10% in each stage. Real-time quantitative PCR (RT-PCR) and dPCR (digital PCR) were used to examine the samples. The conversion factor (CF) was calculated and validated for each laboratory.Results:In the RT-PCR comparison, one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.133-0.338) and 95% limits of agreement within ±5 folds (upper limit 0.147-0.785, lower limit -0.770--0.109), and the corresponding CF values were calculated and validated. In the dPCR comparison, one laboratory did not report results at the second stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.026-0.267) and 95% limits of agreement within±5 folds (upper limit 0.084-0.991, lower limit -0.669--0.135), and the corresponding CF values were calculated and validated. The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%, 0.1%-1%, 1%-10% and >10% could be detected by both RT-PCR and qPCR. When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%, the detection rate of dPCR was higher than that of RT-PCR (85.56% vs. 68.00%).Conclusions:A good consistency is present among various laboratories. The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion. For quantitative detection of BCR::ABL1 p210 deep molecular reaction, dPCR has a higher positive detection rate and more advantages than RT-PCR. To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test, it is imperative for every laboratory to enhance their daily quality control practices.
4.A multicenter study of neonatal stroke in Shenzhen,China
Li-Xiu SHI ; Jin-Xing FENG ; Yan-Fang WEI ; Xin-Ru LU ; Yu-Xi ZHANG ; Lin-Ying YANG ; Sheng-Nan HE ; Pei-Juan CHEN ; Jing HAN ; Cheng CHEN ; Hui-Ying TU ; Zhang-Bin YU ; Jin-Jie HUANG ; Shu-Juan ZENG ; Wan-Ling CHEN ; Ying LIU ; Yan-Ping GUO ; Jiao-Yu MAO ; Xiao-Dong LI ; Qian-Shen ZHANG ; Zhi-Li XIE ; Mei-Ying HUANG ; Kun-Shan YAN ; Er-Ya YING ; Jun CHEN ; Yan-Rong WANG ; Ya-Ping LIU ; Bo SONG ; Hua-Yan LIU ; Xiao-Dong XIAO ; Hong TANG ; Yu-Na WANG ; Yin-Sha CAI ; Qi LONG ; Han-Qiang XU ; Hui-Zhan WANG ; Qian SUN ; Fang HAN ; Rui-Biao ZHANG ; Chuan-Zhong YANG ; Lei DOU ; Hui-Ju SHI ; Rui WANG ; Ping JIANG ; Shenzhen Neonatal Data Network
Chinese Journal of Contemporary Pediatrics 2024;26(5):450-455
Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75%(27/36);boys accounted for 64%(23/36).Among the 36 neonates,31(86%)had disease onset within 3 days after birth,and 19(53%)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61%)had left cerebral infarction and 13(36%)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75%)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72%)and seizures in 10 neonates(34%).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33%,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44%(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.
5.Expression of Trichinella spiralis trehalase(TsTRE)protein and establishment of an indirect ELISA
Jia-Ming HOU ; Bo-Han ZHANG ; Ru-Lin RUAN ; Jian-Hui ZHANG ; Kun LIU ; Hao-Yuan CHEN ; Ming-Xin SONG ; Zi-Qun ZHANG
Chinese Journal of Zoonoses 2023;39(12):1174-1180
We established an indirect ELISA method using Trichinella spiralis trehalase(TsTRE)protein expressed in prokaryotic cells.The TsTRE gene was amplified by RT-PCR and ligated into the pCold I plasmid,which was expressed in E.coli BL21 competent cells.The rTsTRE protein was purified through affinity column chromatography.The TsTRE protein was localized with immunofluorescence techniques,and the immunogenicity of rTsTRE was detected by westernblotting.Subse-quently,rTsTRE protein was used as a coating antigen to establish an indirect ELISA.We optimized the antigen-coating con-centration,serum dilution concentration,antigen-coating incubation time,type of blocking solution,blocking incubation time,HRP-labeled goat anti-rabbit IgG serum dilution concentration,HRP-labeled goat anti-rabbit IgG serum incubation time and response time of TMB.Subsequently,the critical value,repeatability,sensitivity,specificity and clinical detection rate of the ELISA were evaluated.Immunofluorescence indicated that trehalase was abundant in the rod-shaped body,tail and epidermis of Trichinella spiralis muscle larvae.Western-blot indicated that rTsTRE protein combined with the positive serum of mice infected with T.spiralis for 42 d;the band was approximately 60 kDa.The established indirect ELISA had a positive threshold of 0.384;the intra-run and inter-run coefficients of variation were 5.504%-7.630% and 4.664%-9.929%,and did not exceed 10%.The lowest detectable titer was 1:1 280.No cross reaction was observed with antibodies to Clonorchissinensis,Schistosoma ja-ponicum,Ascaris suum,Toxocara gondii and Toxocara canis,and the clinical negative detection rate was 0%.Thus,we suc-cessfully expressed the rTsTRE protein.Moreover,the established indirect ELISA method using the TsTRE protein as the coating antigen had good repeatability,sensitivity,specificity and clinical detectability,and can be applied to the detection of clinical samples.
7.A prospective randomized controlled study on the effects of bicycle ergometer rehabilitation training on quadriceps and walking ability of patients with lower limb dysfunction caused by extensive burns.
Kun Ping WU ; Pei CHEN ; Tian Feng RU ; Lin YUAN ; Hao LUO ; Wei Guo XIE
Chinese Journal of Burns 2022;38(5):447-453
Objective: To explore the effects of bicycle ergometer rehabilitation training on quadriceps and walking ability of patients with lower limb dysfunction caused by extensive burns. Methods: A prospective randomized controlled study was conducted. A total of 40 patients with extensive burns who met the inclusion criteria and were admitted to Tongren Hospital of Wuhan University&Wuhan Third Hospital from December 2017 to December 2020 were selected. According to the random number table, the patients were divided into conventional training group (16 males, 4 females, aged (45±10) years) and combined training group (13 males, 7 females, aged (39±8) years). Patients in conventional training group were given conventional rehabilitation therapy such as joint loosening, lower limb strength training, walking training, and pressure therapy, while patients in combined training group were given additional bicycle ergometer rehabilitation training on the basis of conventional rehabilitation. For patients in the 2 groups before and after a 2-month's treatment, the thickness of quadriceps was measured by ultrasonic diagnostic instrument, the muscle strength of quadriceps was measured by portable muscle strength tester, the walking ability was tested with a 6-min and a 10-meter walk tests, and the patients' satisfaction for treatment effects was assessed using the modified Likert scale. Data were statistically analyzed with independent or paired sample t test, Mann-Whitney U test, Wilcoxon signed rank test, or chi-square test. Results: After 2-month's treatment, the quadriceps thickness of patients in combined training group was (3.76±0.39) cm, which was significantly thicker than (3.45±0.35) cm in conventional training group (t=2.67, P<0.05); quadriceps thickness of patients in conventional training group and combined training group after 2-month's treatment was significantly thicker than that before treatment (with t values of 5.99 and 8.62, respectively, P<0.01). After 2-month's treatment, the quadriceps muscle strength of patients in combined training group was significantly greater than that in conventional training group (Z=2.69, P<0.01); quadriceps muscle strength of patients in conventional training group and combined training group after 2-month's treatment was significantly greater than that before treatment (with Z values of 3.92 and 3.92, respectively, P<0.01). After 2-month's treatment, the 6-min walking distance of patients in combined training group was (488±39) m, which was significantly longer than (429±25) m in conventional training group (t=5.66, P<0.01); the 6-min walking distance of patients after 2-month's treatment in conventional training group and combined training group was significantly longer than that before treatment (with t values of 13.16 and 17.92, respectively, P<0.01). After 2-month's treatment, the 10-meter walking time of patients in combined training group was significantly shorter than that in conventional training group (t=3.20, P<0.01); and the 10-meter walking time in conventional training group and combined training group was significantly shorter than that before treatment (with t values of 7.21 and 13.13, respectively, P<0.01). The patients' satisfaction score for treatment effects in combined training group was significantly higher than that in conventional training group (Z=3.14, P<0.01), and the patients' satisfaction scores for treatment effects in conventional training group and combined training group after 2-month's treatment were significantly greater than those before treatment (with Z values of 3.98 and 4.04, respectively, P<0.01). Conclusions: Bicycle ergometer rehabilitation training can be used to improve quadriceps thickness, muscle strength, and walking ability of patients with lower limb dysfunction caused by extensive burns. It can also improve the satisfaction of patients with the treatment outcome, and therefore is worthy of promotion.
Bicycling
;
Burns/therapy*
;
Female
;
Humans
;
Lower Extremity
;
Male
;
Prospective Studies
;
Quadriceps Muscle
;
Treatment Outcome
;
Walking
8.Analysis on cancer incidence and mortality attributed to human papillomavirus infection in China, 2016.
Mei Wen YUAN ; Hong Hao WANG ; Ru Fei DUAN ; Kun Peng XU ; Shang Ying HU ; You Lin QIAO ; Yong ZHANG ; Fang Hui ZHAO
Chinese Journal of Epidemiology 2022;43(5):702-708
Objective: We aim to evaluate the morbidity and mortality of cancer attributable to human papillomavirus (HPV) infection in China in 2016. Methods: Based on the cancer incidence and mortality rates, national population data, and population attributable fraction (PAF) in China, we calculated the number of incidence and death cases attributed to HPV infection in different areas, age groups, and gender in China in 2016. The standardized incidence and mortality rates for cancer attributed to HPV infection were calculated by using Segi's population. Results: In 2016, a total of 124 772 new cancer cases (6.32 per 100 000) were attributed to HPV infection in China, including 117 118 cases in women and 7 654 cases in men. Of these cancers, cervical cancer was the most common one, followed by anal cancer, oropharyngeal cancer, penile cancer, vaginal cancer, laryngeal cancer, oral cancer, and vulvar cancer. A total of 41 282 (2.03 per 100 000) deaths were attributed to HPV infection, of which 37 417 occurred in women and 3 865 in men. Most deaths were caused by cervical cancer, followed by anal cancer, oropharyngeal cancer, penile cancer, laryngeal cancer, vaginal cancer, oral cancer, and vulvar cancer. The incidence and mortality rates of cervical cancer increased rapidly with age, peaked in age group 50-54 years, then decreased obviously. The morbidity and mortality rates of non-cervical cancer increased with age. The cancer case and death numbers in rural areas (57 089 cases and 19 485 deaths) were lower than those in urban areas (67 683 cases and 21 797 deaths). However, the age-standardized incidence rate (ASIR) and age-standardized mortality rate (ASMR) of cervical cancer were higher in rural areas than in urban areas. There were no significant differences in ASIR and ASMR of non-cervical cancers between urban areas and rural areas. Conclusions: The incidence of cancers attributed to HPV infection in China was lower than the global average, but the number of incidences accounted largely, furthermore there is an increasing trend of morbidity and mortality. The preventions and controls of cervical cancer and male anal cancer are essential to contain the increases in cancer cases and deaths attributed to HPV infection.
China/epidemiology*
;
Female
;
Humans
;
Incidence
;
Laryngeal Neoplasms
;
Male
;
Middle Aged
;
Mouth Neoplasms
;
Oropharyngeal Neoplasms/epidemiology*
;
Papillomavirus Infections/epidemiology*
;
Penile Neoplasms/epidemiology*
;
Registries
;
Uterine Cervical Neoplasms/epidemiology*
;
Vaginal Neoplasms
;
Vulvar Neoplasms
9.Effects of Bushen Yiqi Huoxue Decoction in Treatment of Patients with Diminished Ovarian Reserve: A Randomized Controlled Trial.
Jing ZHOU ; Xin-Yao PAN ; Jin LIN ; Qi ZHOU ; Li-Kun LAN ; Jun ZHU ; Ru DUAN ; Lan WANG ; Yan SUN ; Ling WANG
Chinese journal of integrative medicine 2022;28(3):195-201
OBJECTIVE:
To explore the therapeutic effect of Bushen Yiqi Huoxue Decoction BYHD) in patients with diminished ovarian reserve (DOR).
METHODS:
A total of 180 patients with DOR diagnosed from December 2013 to December 2014 were equally assigned into progynova and duphaston (E+D) group, Zuogui Pill group and BYHD group with 60 cases in each by computerized randomization. Patients received E+D, Zuogui Pill or BYHD for 12 months, respectively. Follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), antral follicle count (AFC), ovarian volume, endometrial thickness, and the resistance indices (RIs) of ovarian arteries and uterine arteries were observed before and after treatment.
RESULTS:
Nine women (4 from the E+D group, 3 from the Zuogui Pill group, and 2 from the BYHD group) withdrew from the study. After 6 months, Zuogui Pill and BYHD significantly decreased FSH and LH and increased endometrial thickness and AMH (all P<0.01). BYHD also resulted in E2 elevation (P<0.05), ovary enlargement (P<0.05), AFC increase (P<0.01), and RI of ovarian arteries decrease (P<0.05). After 12 months, further improvements were observed in the Zuogui Pill and BYHD groups (all P<0.01), but BYHD showed better outcomes, with lower FSH, larger ovaries and a thicker endometrium compared with the Zuogui Pill group (all P<0.01). However, E+D only significantly increased endometrial thickness (P<0.01) and no significant improvements were observed in the RI of uterine arteries in the three groups.
CONCLUSIONS
BYHD had a favorable therapeutic effect in patients with DOR by rebalancing hormone levels, promoting ovulation, and repairing the thin endometrium. The combination of tonifying Shen (Kidney), benefiting qi and activating blood circulation may be a promising therapeutic strategy for DOR.
Anti-Mullerian Hormone/pharmacology*
;
Drugs, Chinese Herbal
;
Female
;
Follicle Stimulating Hormone
;
Humans
;
Luteinizing Hormone
;
Ovarian Reserve
10.Characteristics of fusion gene expression in acute lymphoblastic leukemia.
Xian Qi HUANG ; Ya Ni LIN ; En Bin LIU ; Fei XING ; Zhe WANG ; Xue Jing CHEN ; Long CHEN ; Jing Ting MA ; Ying Chang MI ; Kun RU
Chinese Journal of Pathology 2022;51(4):307-313
Objective: To analyze the genetic landscape of 52 fusion genes in patients with de novo acute lymphoblastic leukemia (ALL) and to investigate the characteristics of other laboratory results. Methods: The fusion gene expression was retrospectively analyzed in the 1 994 patients with de novo ALL diagnosed from September 2016 to December 2020. In addition, their mutational, immunophenotypical and karyotypical profiles were investigated. Results: In the 1 994 patients with ALL, the median age was 12 years (from 15 days to 89 years). In the panel of targeted genes, 15 different types of fusion genes were detected in 884 patients (44.33%) and demonstrated a Power law distribution. The frequency of detectable fusion genes in B-cell ALL was significantly higher than that in T-cell ALL (48.48% vs 18.71%), and fusion genes were almost exclusively expressed in B-cell ALL or T-cell ALL. The number of fusion genes showed peaks at<1 year, 3-5 years and 35-44 years, respectively. More fusion genes were identified in children than in adults. MLL-FG was most frequently seen in infants and TEL-AML1 was most commonly seen in children, while BCR-ABL1 was dominant in adults. The majority of fusion gene mutations involved signaling pathway and the most frequent mutations were observed in NRAS and KRAS genes. The expression of early-stage B-cell antigens varied in B-cell ALL patients. The complex karyotypes were more common in BCR-ABL1 positive patients than others. Conclusion: The distribution of fusion genes in ALL patients differs by ages and cell lineages. It also corresponds to various gene mutations, immunophenotypes, and karyotypes.
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Child
;
Child, Preschool
;
Gene Expression
;
Genes, ras
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Humans
;
Infant
;
Infant, Newborn
;
Middle Aged
;
Oncogene Fusion
;
Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism*
;
Retrospective Studies
;
Young Adult

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