1.Lineage tracing for mammary stem cells using Hopx reporter mice
Jianyun SHI ; Wenjing LI ; Ying PENG ; Zhenhua JIA ; Shujin ZHANG ; Lulu TAN ; Yitong YUAN ; Ruochen DU
Chinese Journal of Tissue Engineering Research 2025;29(13):2720-2727
BACKGROUND:Mammary stem cells are vital for the development and homeostasis of mammary gland tissue.The occurrence of breast cancer has a close relationship with the mammary stem cells.Recent studies have shown that Hopx,as an important transcriptional regulator of morphogenesis and cell differentiation,has been confirmed to be expressed in a variety of adult stem cells such as nerves,intestines,hair follicles and lungs.However,its role in mammary stem cells has not been reported so far.OBJECTIVE:To investigate whether Hopx expression marks mammary stem cells.METHODS:(1) Female Hopx-LacZ transgenic mice aged 8 weeks were selected to detect the background expression of Hopx in breast tissue by β-galactosidase staining.(2) Female wild-type mice at 4,6,and 8 weeks of age and 14.5 days of gestation were selected for whole-tissue magenta staining and K14 and K8 immunofluorescence staining,respectively.(3) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks and 17.5 days of gestation were selected and stained with breast β-galactosidase.(4) The 4-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 weeks after injection.The 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 and 10 weeks after the last injection.(5) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times).Hopx-CreERT2;Rosa26LacZ transgenic mice were pregnant 2 weeks after injection.The mammary tissue of mice at 17.5 days of the first pregnancy and 17.5 days of the third pregnancy was stained with β-galactosidase.RESULTS AND CONCLUSION:(1) The results of β-galactosidase staining showed that the mammary ducts of Hopx-LacZ transgenic mice at 8 weeks of age did contain Hopx-positive cells and were located in the basal epithelia,with a small number.(2) Whole-mount staining of mammary glands and immunofluorescence staining results exhibited that the mammary glands of mice had different characteristics with corresponding developmental stages such as puberty,maturity,and pregnancy,and underwent a series of complex epithelial remodeling processes.(3) The results of β-galactosylase staining showed that Hopx-labeled positive cells in the mammary duct of Hopx-CreERT2;Rosa26LacZ transgenic mice at 17.5 days of gestation increased compared with female Hopx-CreERT2;Rosa26LacZ transgenic mice at 8 weeks of age.(4) The results of β-galactosylase staining showed that the Hopx-labeled positive cells in the mammary glands of 4-and 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice after tamoxifen injection were located in the basal epithelium with a small number.(5) The results of β-galactosidase staining showed that Hopx-labeled positive cells in the mammary glands of mice at 17.5 days of the first and third gestation were located in the basal epithelia around the alveoli,and the number of Hopx-labeled positive cells at 17.5 days of the third gestation was more.(6) In conclusion,Hopx reporter-marked basal epithelial cells belong to dormant mammary stem cells,which are responsible for the growth of the mammary glands during pregnancy and contribute to acinar formation.
2.Lineage tracing for mammary stem cells using Hopx reporter mice
Jianyun SHI ; Wenjing LI ; Ying PENG ; Zhenhua JIA ; Shujin ZHANG ; Lulu TAN ; Yitong YUAN ; Ruochen DU
Chinese Journal of Tissue Engineering Research 2025;29(13):2720-2727
BACKGROUND:Mammary stem cells are vital for the development and homeostasis of mammary gland tissue.The occurrence of breast cancer has a close relationship with the mammary stem cells.Recent studies have shown that Hopx,as an important transcriptional regulator of morphogenesis and cell differentiation,has been confirmed to be expressed in a variety of adult stem cells such as nerves,intestines,hair follicles and lungs.However,its role in mammary stem cells has not been reported so far.OBJECTIVE:To investigate whether Hopx expression marks mammary stem cells.METHODS:(1) Female Hopx-LacZ transgenic mice aged 8 weeks were selected to detect the background expression of Hopx in breast tissue by β-galactosidase staining.(2) Female wild-type mice at 4,6,and 8 weeks of age and 14.5 days of gestation were selected for whole-tissue magenta staining and K14 and K8 immunofluorescence staining,respectively.(3) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks and 17.5 days of gestation were selected and stained with breast β-galactosidase.(4) The 4-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 weeks after injection.The 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 and 10 weeks after the last injection.(5) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times).Hopx-CreERT2;Rosa26LacZ transgenic mice were pregnant 2 weeks after injection.The mammary tissue of mice at 17.5 days of the first pregnancy and 17.5 days of the third pregnancy was stained with β-galactosidase.RESULTS AND CONCLUSION:(1) The results of β-galactosidase staining showed that the mammary ducts of Hopx-LacZ transgenic mice at 8 weeks of age did contain Hopx-positive cells and were located in the basal epithelia,with a small number.(2) Whole-mount staining of mammary glands and immunofluorescence staining results exhibited that the mammary glands of mice had different characteristics with corresponding developmental stages such as puberty,maturity,and pregnancy,and underwent a series of complex epithelial remodeling processes.(3) The results of β-galactosylase staining showed that Hopx-labeled positive cells in the mammary duct of Hopx-CreERT2;Rosa26LacZ transgenic mice at 17.5 days of gestation increased compared with female Hopx-CreERT2;Rosa26LacZ transgenic mice at 8 weeks of age.(4) The results of β-galactosylase staining showed that the Hopx-labeled positive cells in the mammary glands of 4-and 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice after tamoxifen injection were located in the basal epithelium with a small number.(5) The results of β-galactosidase staining showed that Hopx-labeled positive cells in the mammary glands of mice at 17.5 days of the first and third gestation were located in the basal epithelia around the alveoli,and the number of Hopx-labeled positive cells at 17.5 days of the third gestation was more.(6) In conclusion,Hopx reporter-marked basal epithelial cells belong to dormant mammary stem cells,which are responsible for the growth of the mammary glands during pregnancy and contribute to acinar formation.
3.Microbiological surveillance result of endoscopes after INTERCEPT Foam Spray: a quasi-experimental pilot study in Singapore
Caihong WANG ; Rong ZHANG ; Ruhui FAN ; Jiewen LOW ; Ruochen DU ; Xueyun MA ; Congcong CAI
Clinical Endoscopy 2024;57(6):821-831
Background/Aims:
This study aimed to assess the impact of INTERCEPT Foam Spray (IFS) application on delayed endoscope reprocessing through microbiological surveillance culture (MSC).
Methods:
A quasi-experimental, matched-comparison pilot study was conducted using gastrointestinal endoscopy. IFS was applied to the endoscopes after precleaning and before reprocessing the next day. An equal number of endoscopes, matched by endoscope type, were subjected to routine reprocessing. The MSC were subjected to high-level disinfection to detect any contamination. Data were analyzed using the chi-square test or Fisher exact test (categorical data) and Student t-test (continuous data).
Results:
In total, 150 MSCs were collected from 42 endoscopes. Positive MSCs were observed in 4.0% (4/75) of the sprayed group and 1.3% (1/75) of the control group (95% confidence interval, 30.34–0.31; p>0.05), all of which were contributed by colonoscopes. Colonoscope were more prone to positive MSC (mean difference in percentage, p<0.05). Mean spraying hours were not associated with detected growth (11.7% vs. 13.6%; 95% confidence interval, 1.43 to –5.27; p>0.05), with environmental and skin flora being the primary contaminants.
Conclusions
IFS may be applied when delayed endoscope processing is necessary, but with caution when applied to colonoscopes. However, further research is warranted to verify the result.
4.Microbiological surveillance result of endoscopes after INTERCEPT Foam Spray: a quasi-experimental pilot study in Singapore
Caihong WANG ; Rong ZHANG ; Ruhui FAN ; Jiewen LOW ; Ruochen DU ; Xueyun MA ; Congcong CAI
Clinical Endoscopy 2024;57(6):821-831
Background/Aims:
This study aimed to assess the impact of INTERCEPT Foam Spray (IFS) application on delayed endoscope reprocessing through microbiological surveillance culture (MSC).
Methods:
A quasi-experimental, matched-comparison pilot study was conducted using gastrointestinal endoscopy. IFS was applied to the endoscopes after precleaning and before reprocessing the next day. An equal number of endoscopes, matched by endoscope type, were subjected to routine reprocessing. The MSC were subjected to high-level disinfection to detect any contamination. Data were analyzed using the chi-square test or Fisher exact test (categorical data) and Student t-test (continuous data).
Results:
In total, 150 MSCs were collected from 42 endoscopes. Positive MSCs were observed in 4.0% (4/75) of the sprayed group and 1.3% (1/75) of the control group (95% confidence interval, 30.34–0.31; p>0.05), all of which were contributed by colonoscopes. Colonoscope were more prone to positive MSC (mean difference in percentage, p<0.05). Mean spraying hours were not associated with detected growth (11.7% vs. 13.6%; 95% confidence interval, 1.43 to –5.27; p>0.05), with environmental and skin flora being the primary contaminants.
Conclusions
IFS may be applied when delayed endoscope processing is necessary, but with caution when applied to colonoscopes. However, further research is warranted to verify the result.
5.Microbiological surveillance result of endoscopes after INTERCEPT Foam Spray: a quasi-experimental pilot study in Singapore
Caihong WANG ; Rong ZHANG ; Ruhui FAN ; Jiewen LOW ; Ruochen DU ; Xueyun MA ; Congcong CAI
Clinical Endoscopy 2024;57(6):821-831
Background/Aims:
This study aimed to assess the impact of INTERCEPT Foam Spray (IFS) application on delayed endoscope reprocessing through microbiological surveillance culture (MSC).
Methods:
A quasi-experimental, matched-comparison pilot study was conducted using gastrointestinal endoscopy. IFS was applied to the endoscopes after precleaning and before reprocessing the next day. An equal number of endoscopes, matched by endoscope type, were subjected to routine reprocessing. The MSC were subjected to high-level disinfection to detect any contamination. Data were analyzed using the chi-square test or Fisher exact test (categorical data) and Student t-test (continuous data).
Results:
In total, 150 MSCs were collected from 42 endoscopes. Positive MSCs were observed in 4.0% (4/75) of the sprayed group and 1.3% (1/75) of the control group (95% confidence interval, 30.34–0.31; p>0.05), all of which were contributed by colonoscopes. Colonoscope were more prone to positive MSC (mean difference in percentage, p<0.05). Mean spraying hours were not associated with detected growth (11.7% vs. 13.6%; 95% confidence interval, 1.43 to –5.27; p>0.05), with environmental and skin flora being the primary contaminants.
Conclusions
IFS may be applied when delayed endoscope processing is necessary, but with caution when applied to colonoscopes. However, further research is warranted to verify the result.
6.Gender-Specific Reference Charts of Fetal Head Circumference in a Singaporean Population.
George Sh YEO ; Maili QI ; Ruochen DU ; Padma Lata MAHAVADI ; Chee Fu YUNG ; Koh Cheng THOON ; Edwin Wh THIA ; Kai Lit TAN ; Fon Min LAI ; Nicole Kl LEE
Annals of the Academy of Medicine, Singapore 2017;46(10):367-373
INTRODUCTIONWith the global outbreak of Zika virus and its association with microcephaly, an up-to-date fetal head circumference (HC) nomogram is crucial to offer a reference standard in order to make an accurate diagnosis. This study was conducted to revise the local fetal HC nomogram.
MATERIALS AND METHODSIn this retrospective study, ultrasound data was used for construction of the fetal HC nomogram from a total of 6155 pregnancies in the ethnic Chinese population with low risk profile at KK Women's and Children's Hospital over a 10-year period. Regression model was fitted to calculate the mean and standard deviation of HC at each gestational age (GA). Comparison of HC between ethnic groups (no significant differences) and genders were made. The revised chart was compared with another commonly used reference chart (Hadlock). In an independent test population, different reference charts were used to estimate number of cases with microcephaly.
RESULTSA statistically significant difference of HC between the genders was observed across all gestational ages. Gender-specific reference charts and equation were computed. Our revised fetal HC chart showed a different distribution from the Hadlock chart. Compared with the gender-specific charts, the Hadlock HC chart would significantly under-report microcephaly cases in male fetuses, and tend to over-report in female fetuses.
CONCLUSIONThis study provides a new set of gender-specific fetal HC charts in the Singaporean population for antenatal ultrasound surveillance of microcephaly.
7.Clinical observation on rheumatoid arthritis treated with modified Duhuo-Jisheng decoction and Western conventional treatment
Youming XIE ; Yanjie FENG ; Ruochen DU ; Han HUANG
International Journal of Traditional Chinese Medicine 2014;(8):685-688
Objective To evaluate the effects of rheumatoid arthritis treated with modified Duhuo-Jisheng decoction and western conventional treatment. Methods 120 patients with heumatoid arthritis of our hospital from January 2010 to 2013 August were recruited and randomly divided into a treatment group and a control group, 60 patients in each group. The control group was treated with oral intake of Methotrexate, Sulfasalazine and Meloxicam in acute stage and oral intake of diclofenc in stable stage, while the treatment group was further added modified Duhuo-Jisheng decoction on the basis of the control group. TCM symptom score, arthritis symptom score, symptomatic relief time, ESR, CRP, RF were observed after 3 months’ treatment. Results After 3 mouths’ treatment, the total effective rate was 90.0%(54/60) in the study group and 70.0%(42/64) in the control group. There was no significant difference(χ2=4.193,P<0.05) between the two groups. CRP,ESR, RF[The treatment group respectively(22.06±10.31)mg/L, (25.18±17.80)mm/h, (28.19±4.17)IU/L,the control group respectively (14.11±7.32)mg/L, (24.16±22.09)mm/h, (36.08±5.24)IU/L]decreased after the treatment in both groups compared with the values before the treatment[The treatment group respectively (82.16± 21.37)mg/L, (68.84±9.71)mm/h, (84.92±15.31)IU/L,the control group respectively (52.46±22.26)mg/L, (62.72±33.31)mm/h, (85.17±14.23)IU/L, P<0.01],while the decrease of CRP in the treatment group was much obvious than the control group(P<0.05),but the change of ESR had no statistical significance between the two group after the treatment(P>0.05);Number of joint tenderness, The number of swollen joints, Time of morning stiffness, Dual hands grip strength , 20 m walking time decreased obviously in both groups after the treatment[After the treatment, the treatment group respectively (3.43 ± 1.46), (2.95 ± 1.35), (19.32 ± 16.54)min,(79.35±22.14)mmHg,(19.32±4.81)s,the control group respectively (4.63±4.21), (3.55±2.47), (21.23 ± 19.37)min,(77.81 ± 16.22)mmHg,(25.15 ± 5.81)s before the treatment, the treatment group respectively(10.71±5.12), (7.95±3.97), (109.32±68.52)min, (47.12±18.22)mmHg, (36.21±6.62)s, the control group respectively (11.54±5.32), (7.64±4.01), (96.02±58.39)min, (49.67±16.48)mmHg, (36.33± 7.23)s](P<0.05). Conclusion Duhuo-Jisheng decoction could effectively improve the clinical symptoms of aged patients with osteoporosis and reduce the levels of their ESR, CRP, and RF.
8.Effects of Strengthening Internal and External Abdominal Oblique Muscles on Sitting Balance in Stroke Patients with Hemiplegia
Zhongbai MA ; Bin HAN ; Ruochen DU
Chinese Journal of Rehabilitation Theory and Practice 2010;16(11):1042-1043
ObjectiveTo investigate the effects of strengthening internal and external abdominal oblique muscles on sitting balance in stroke patients with hemiplegia. Methods60 cases of patients with hemiplegia were randomly divided into treatment group and control group, 30 cases in each group. Control group patients were treated with conventional treatment. Internal and external abdominal oblique muscles training were accepted in treatment group in addition. Before and 1 month after treatment, all the patients were assessed with Fugl-Meyer Assessment of balance function and muscle strength. ResultsAfter treatment, the Fugl-Meyer assessment scores of balance, muscle strength were significantly higher in the treatment group than in the control group(P<0.01). ConclusionMuscle strengthen training in internal and external abdominal oblique muscles will promote sitting balance in hemiplegic patients.


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