1.Establishment of SHERLOCK-HBA Detection Method and Its Application in Blood Identification
Qian-Wei YAO ; Hong-Xia HE ; Sheng HU ; Yi-Xia ZHAO ; Yu LUO ; An-Quan JI ; Qi-Fan SUN
Progress in Biochemistry and Biophysics 2024;51(8):1971-1982
ObjectiveRapid and accurate identification of body fluid traces at crime scenes is crucial for case investigation. Leveraging the speed and sensitivity of nucleic acid detection technology based on SHERLOCK, our research focuses on developing a peripheral blood SHERLOCK-HBA detection system to detect mRNA in forensic practice. MethodsShort crRNA fragments targeting the blood-specific mRNA gene HBA were designed and screened, alongside RPA primers. Optimal RPA primers were selected based on specificity and amplification efficiency, leading to the establishment of the RPA system. The most efficient crRNA was chosen based on relative fluorescence units (RFU) generated by the Cas protein reaction, and the Cas protein reaction system was constructed to establish the SHERLOCK-HBA detection method. The RPA and Cas protein reaction systems in the SHERLOCK detection system were then individually optimized. A total of 79 samples of five body fluids were tested to evaluate the method’s ability to identify blood, with further verification through species-specific tests, sensitivity tests, mixed spots detection, aged samples, UV-irradiated samples, and actual casework samples. ResultsThe SHERLOCK reaction system for the peripheral blood-specific marker HBA was successfully established and optimized, enabling detection within 30 min. The method demonstrated a detection limit of 0.001 ng total RNA, better than FOB strip method and comparable to RT-PCR capillary electrophoresis. The system could detect target body fluids in mixed samples and identify blood in samples stored at room temperature for three years and exposed to UV radiation for 32 h. Detection of 11 casework samples showed performance comparable to RT-PCR capillary electrophoresis. ConclusionThis study presents a CRISPR/Cas-based SHERLOCK-HBA detection system capable of accurately, sensitively, and rapidly identifying blood samples. Introducing CRISPR/Cas technology to forensic body fluid identification represents a significant advancement in applying cutting-edge molecular biology techniques to forensic science.The method’s simplicity, shorter detection time, and independence from specialized equipment make it promising for rapid blood sample identification in forensic cases.
2.Discussion on TCM Etiology and Pathogenesis of Cognitive Dysfunction from the of Perspective Microbiota-gut-brain Axis Based on"Spleen Can't Regulate Spirit"
Lanhui ZHENG ; Qi ZHANG ; Boyu ZHANG ; Dandan FENG ; Jiayao LUO ; Tong YANG ; Hua SUI ; Yan WANG ; Qingquan SUN ; Sheng LI ; Shuyuan LIU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(6):18-23
Cognitive dysfunction refers to dysfunction of individual perception,memory,understanding,learning,creation and other dysfunctions caused by abnormal brain function and structure.Based on the fact that the spleen can't regulate transportation and transformation,govern blood and send up essential substance,combined with the microbiota-gut-brain axis,this article discussed the etiology and pathogenesis of intestinal flora imbalance affecting cognitive dysfunction in TCM.It was proposed that the spleen in TCM and intestinal flora are connected in physiology and pathology:the spleen regulates spirit and governs cognition,when the spleen fails to function normally that it can't dominate transportation and transformation,govern blood and send up essential substance will cause that the brain spirit can not be nourished;intestinal flora is closely related to the spleen in TCM,and affects brain function through the nervous system,endocrine,immune and metabolic mechanisms.This article can provide explore new ideas for the clinical research and treatment of cognitive dysfunction of traditional Chinese and Western medicine.
3.Development and characteristics of auricular fumigation moxibustion combined with heat-sensitive moxibustion device.
Xue-Tao ZHANG ; Zhen WANG ; Han-Xiao WANG ; Ya-Lu WANG ; Xin WU ; Kun WANG ; Shuai CUI ; Mei-Qi ZHOU ; Hui LUO ; Can-Guang SUN ; Sheng-Bing WU
Chinese Acupuncture & Moxibustion 2023;43(11):1333-1337
A moxibustion device with the functions of auricular fumigation moxibustion and heat-sensitive moxibustion is designed. The smoke of the ignited moxa stick is used for the fumigation moxibustion at the external auditory canal, while the heat generated works on Dazhui (GV 14) for heat-sensitive moxibustion. The device consists of five parts, i.e. combustion chamber, smoke pipe, smoke processing chamber, power module and connector. It solves the limitations such as unpleasant experience in treatment, unfavorable temperature control, easy scalding and excessive manual dependence induced by usual fumigation moxibustion and during heat-sensitive moxibustion. This moxibustion device may improve the safety and convenience when delivering the treatment with fumigation moxibustion and heat-sensitive moxibustion, as well as the work efficiency of medical staff.
Humans
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Moxibustion
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Hot Temperature
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Fumigation
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Smoke
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Temperature
4.Mid-term effectiveness of hip preservation in the reconstruction of ultrashort bone segments in the proximal femur with three-dimensional printed customized cementless intercalary endoprosthesis with an intra-neck curved stem.
Hongtao SHENG ; Yuqi ZHANG ; Qi YOU ; Taojun GONG ; Zhuangzhuang LI ; Xuanhong HE ; Fan TANG ; Yong ZHOU ; Yitian WANG ; Minxun LU ; Yi LUO ; Li MIN ; Chongqi TU
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(8):970-977
OBJECTIVE:
To explore the design points of a three-dimensional (3D) printed customized cementless intercalary endoprosthesis with an intra-neck curved stem and to evaluate the key points and mid-term effectiveness of its application in the reconstruction of ultrashort bone segments in the proximal femur.
METHODS:
Between October 2015 and January 2021, 17 patients underwent reconstruction with a 3D printed-customized cementless intercalary endoprosthesis with an intra-neck curved stem. There were 11 males and 6 females, the age ranged from 10 to 76 years, with an average of 30.1 years. There were 9 cases of osteosarcoma, 4 cases of Ewing sarcoma, 2 cases of chondrosarcoma, 1 case of liposarcoma, and 1 case of myofibroblastoma. The disease duration was 5-14 months, with an average of 9.5 months. Enneking staging included 16 cases of stage ⅡB and 1 case of stage ⅢB. The distances from the center of the femoral head to the body midline and the acetabular apex were measured preoperatively on X-ray images. Additionally, the distances from the tip of the intra-neck curved stem to the body midline and the acetabular apex were measured at immediate postoperatively and last follow-up. The neck-shaft angle was also measured preoperatively, at immediate postoperatively, and at last follow-up. The status of osseointegration at the bone-prosthesis interface and bone growth into the prosthesis surface were assessed by X-ray films, CT, and Tomosynthesis-Shimadzu metal artefact reduction technology (T-SMART). The survival status of the patients, presence of local recurrence or distant metastasis, and occurrence of postoperative complications were assessed. The recovery of lower limb function was evaluated pre- and post-operatively using the Musculoskeletal Tumor Society (MSTS) scoring system, and pain relief was evaluated using the visual analogue scale (VAS) scores.
RESULTS:
The patient's femoral resection length was (163.1±57.5) mm, the remaining proximal femoral length was (69.6±9.3) mm, and the percentage of femoral resection length/total femoral length was 38.7%±14.6%. All 17 patients were followed up 25-86 months with an average of 58.1 months. During the follow-up, 1 patient died of lung metastasis at 46 months postoperatively, and the remaining 16 patients survived tumor-free. There was no complication such as periprosthetic infection, delayed incision healing, aseptic loosening, prosthesis fracture, or periprosthetic fracture. No evidence of micromotion or wear around the implanted stem of the prosthesis was detected in X-ray and T-SMART evaluations. There was no significant radiolucent lines, and radiographic evidence of bone ingrowth into the bone-prosthesis interface was observed in all stems. There was no significant difference in the distance from the tip of the curved stem to the body midline and the apex of the acetabulum at immediate postoperatively and last follow-up compared with the distance from the center of the femoral head to the body midline and the apex of the acetabulum before operation, respectively (P>0.05), and there was no significant difference in the above indexes between immediate postoperatively and last follow-up (P>0.05). The differences in the neck-shaft angle at various time points before and after operation were also not significant (P>0.05). At last follow-up, the MSTS score was 26.1±1.2 and the VAS score was 0.1±0.5, which were significantly improved when compared with those before operation [19.4±2.1 and 5.7±1.0, respectively] (t=14.735, P<0.001; t=21.301, P<0.001). At last follow-up, none of the patients walked with the aid of crutches or other walkers.
CONCLUSION
The 3D printed customized cementless intercalary endoprosthesis with an intra-neck curved stem is an effective method for reconstructing ultrashort bone segments in the proximal femur following malignant tumor resection. The operation is reliable, the postoperative lower limb function is satisfactory, and the incidence of complications is low.
Female
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Male
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Humans
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Child
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Adolescent
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Young Adult
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Adult
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Middle Aged
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Aged
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Femur/surgery*
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Lower Extremity
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Bone-Implant Interface
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Femur Head
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Artificial Limbs
6.Development and characteristics of automatic ash-removal heat-sensitive moxibustion device.
Xue-Tao ZHANG ; Han-Xiao WANG ; Zhen WANG ; Ya-Lu WANG ; Shuai CUI ; Mei-Qi ZHOU ; Hui LUO ; Can-Guang SUN ; Ming HAO ; Wen HONG ; Sheng-Bing WU
Chinese Acupuncture & Moxibustion 2023;43(5):597-599
An automatic ash-removal heat-sensitive moxibustion device was developed, which could keep relatively constant temperature of heat-sensitive moxibustion, and realize the automatic ignition and automatic ash removal of moxa sticks during heat-sensitive moxibustion. The automatic ash-removal heat-sensitive moxibustion device comprises a bracket and a moxibustion box fixed on the top of the bracket; the bracket is composed of a base and a movable telescopic arm. This device can solve the problems of temperature instability, moxa ash blocking heat transfer and moxa ash falling during heat-sensitive moxibustion, avoiding the scalding caused by moxa ash falling, and reduce the workload of medical staff.
Humans
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Hot Temperature
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Moxibustion
;
Temperature
8.Nicotine exacerbates tacrolimus-induced renal injury by programmed cell death
Yu Ji JIANG ; Sheng CUI ; Kang LUO ; Jun DING ; Qi Yan NAN ; Shang Guo PIAO ; Mei Ying XUAN ; Hai Lan ZHENG ; Yong Jie JIN ; Ji Zhe JIN ; Jung Pyo LEE ; Byung Ha CHUNG ; Bum Soon CHOI ; Chul Woo YANG ; Can LI
The Korean Journal of Internal Medicine 2021;36(6):1437-1449
Background/Aims:
Cigarette smoking is an important modifiable risk factor in kidney disease progression. However, the underlying mechanisms for this are lacking. This study aimed to assess whether nicotine (NIC), a major toxic component of cigarette smoking, would exacerbates tacrolimus (TAC)-induced renal injury.
Methods:
Sprague-Dawley rats were treated daily with NIC, TAC, or both drugs for 4 weeks. The influence of NIC on TAC-caused renal injury was examined via renal function, histopathology, oxidative stress, mitochondria, endoplasmic reticulum (ER) stress, and programmed cell death (apoptosis and autophagy).
Results:
Both NIC and TAC significantly impaired renal function and histopathology, while combined NIC and TAC treatment aggravated these parameters beyond the effects of either alone. Increased oxidative stress, ER stress, mitochondrial dysfunction, proinf lammatory and profibrotic cytokine expressions, and programmed cell death from either NIC or TAC were also aggravated by the two combined.
Conclusions
Our observations suggest that NIC exacerbates chronic TAC nephrotoxicity, implying that smoking cessation may be beneficial for transplant smokers taking TAC.
9.Pathological Mechanism of PPs in Immune-related Enteropathy and Regulatory Effect of Traditional Chinese Medicine Polysaccharides
Xiu-lan PU ; Mei-si LIN ; Rui-feng LUO ; Qi-yan CHEN ; Ling-ling DONG ; Da-sheng LIN ; Fei GAO
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(4):216-224
Intestinal Peyer's patches (PPs) are local immune tissues of the intestine, which are considered to be the main induction site of the intestinal mucosal immune response, and closely related to immune-related refractory enteropathies, such as ulcerative colitis and Crohn's disease. In recent years, more and more scholars have tried to find a new breakthrough for treating refractory enteropathies with a limited efficacy of clinical interventions through in-depth study of the relationship between PPs and enteropathy. Traditional Chinese medicine (TCM) polysaccharides are considered to be a key component for immune regulation with TCM. Modern studies show that TCM polysaccharides have a significant positive intervention effect on the structure and function of PPs, with good development prospects. Based on this, this paper focuses on PPs and intestinal-related diseases, and systematically introduces the physiological structure of PPs and their drug delivery mechanism, and summarizes the interactions of PPs with effect on immune-related enteropathies, analyze of current studies and prospects of effect of TCM polysaccharides in intervening intestinal disease and its dysfunction by regulating PPs, with the aim to provide new strategies for basic studies and clinical treatment of immune-related refractory enteropathies from the perspective of PPs, and new ideas for basic studies and clinical studies on effect of TCM polysaccharides in intervening enteropathies.
10.Effect of Removing Core on Quality of Phyllanthi Fructus
Chuan-hong LUO ; Sheng-jie HUANG ; Qi-qi HU ; Peng TAN ; Hua-mei GOU ; Xi-chuan WEI ; Hao-zhou HUANG ; San-hu FAN ; Jun-zhi LIN ; Ding-kun ZHANG ; Li HAN
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(9):147-156
Objective:The differences of chemical compositions and pharmacological activities between the core and pulp of Phyllanthi Fructus were investigated by chemical analysis and

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