1.Three-dimensional culture of marrow mesenchymal stem cells under microgravity on injectable Pluronic F-127 in the repair of articular cartilage defects
Hc WANG ; Zy ZHANG ; Wgfirst XIN
Chinese Journal of Tissue Engineering Research 2007;0(14):-
AIM: To repair the articular cartilage defects with the non-cartilage derived plant cells induced from allogenic mesenchymal stem cells (MSCs) in microgravity dynamic culture system of three-dimensional (3D) Pluronic F-127. METHODS: The experiment was conducted in the Experimental Animal Center, First Affiliated Hospital of Harbin Medical University from June 2005 to March 2006. ①Twenty-seven two-month New Zealand rabbits were selected, of which 3 were used as the donors of mesenchymal stem cells, and the left rabbits were randomly divided into induced cell and Pluronic group, pluronic group and control group with 8 rabbits (16 knees) in each group. Pluronic F-127 (Sigma Company, No. P2443) was white power with no odour, and its water solution was liquidness at 4 ℃, and solid gelatum at 37 ℃. ②After the rabbits were anesthetized, the bone marrow was taken from superior extremity of tibia to isolate and culture the MSCs; the 3rd passage cells were used in the experiment. ③The MSCs were collected and centrifuged, then divided into four groups with different inducing conditions: 3D dynamic culture group: The cells were washed with 10 g/L algin solution and renewed to suspend cells with density of 5?1010 L-1; 200 mmol/L calcium chloride solution was dropped into and calcium alginate gelatum microballoons formed immediately, and the cells were floated and fixed in the microballoons. Five minutes later, the gelatum microballoons were dislodged and put in Revolve Cell Culture System (RCCS) for dynamic culture under microgravity. 2D dynamic culture group: The cells were given inoculation directly in RCCS for dynamic culture under microgravity. 3D static culture group: The cells in the calcium alginate gelatum were inoculated directly in a culture flask for static culture. 2D static culture group: The cells were inoculated directly in a plane culture flask for static culture. Each group was cultured for 2 weeks, following by slices preparation and toluidine orchid dyeing, collogenⅠ, Ⅱ immunohistochemical staining. Citrate sodium solution was adopted to dissolve the calcium alginate gelatum microballoons to determine the collogen and proteoglycan contents of the four groups. ④The models of full-thickness defects of articular cartilage were prepared. Induced cells and Pluronic group: The best induced MSCs compounded with 25% Pluronic F-127 were mixed at 4℃ and suspended, until the final concentration of MSCs was 5?1010 L-1, which poured into the articular cartilage defects. Pluronic group: 25% Pluronic F-127 was poured into the articular cartilage defects. The articular cartilage defects of the control group were not given any treatment. The animals of each group were killed at 4, 8, 12 and 24 weeks to observe the repair effect of the defects grossly and microscopically. Meanwhile, the histological examinations were performed.RESULTS: All 24 animals entered the result analysis. ①The results of culture and examination of MSCs: Algin contacted Ca2+ formed quickly transparent gelatum microballoons, which was lustered; the cells in the microballoons were globular, with clear nucleoles, and the stereochemical structure was similar with normal chondrocyte. Toluidine blue dyeing was positive, and expressed collagen Ⅱ but no collagenⅠwas found. ②Expressions of cell biochemical indicator of the four groups under different inducing conditions: Collagen Ⅱ and proteoglycan production of the 3D and 2D dynamic culture groups were higher than the two static culture groups, and the 3D dynamic culture group was the best (0.078?0.002), (0.048?0.002), (0.035?0.001) A, P
2.Medication combined with local hyperthermia: a desirable therapy for chronic prostatitis pain symptoms.
Yong ZHANG ; Feng-Ling SUN ; Tong ZANG
National Journal of Andrology 2008;14(5):451-453
OBJECTIVETo compare the effect of the combined therapy of medication with local hyperthermia with that of the simple local hyperthermia therapy in the treatment of pain symptoms of chronic prostatitis (CP).
METHODSSeventy-six CP patients aged 18-48 (mean 29.2 +/- 3.8) years, with the disease course of 3.5-180 (mean 8.0 +/- 1.2) months and NIH-CPSI pain score > or = 14, were equally randomized into a treatment group and a control. The former was treated by applying the CRS-2280E extraorgan short-wave capacitance field hyperthermia system to the prostate once an hour every other day for 7 times, combined with anal administration of 1 Qianliean suppository and oral doxazosin 4 mg before bedtime every night for 2 weeks, while the latter underwent simple local hyperthermia. All the patients were scored on NIH-CPSI and the therapeutic results were compared between the two groups.
RESULTSThe pre- and post-treatment NIH-CPSI scores were (23.9 +/- 3.8) and (5.2 +/- 3.1) (P < 0.01) in the treatment goup and (24.5 +/- 4.3) and (11.6 +/- 3.4) (P < 0.01) in the control; the pre- and post-treatment scores on NIH-CPSI pain symptoms were (16.5 +/- 1.9) and (3.1 +/- 2.2) (P < 0.01) in the former and (15.9 +/- 1.7) and (8.2 +/- 2.0) (P < 0.01) in the latter. The total score on NIH-CPSI and that on NIH-CPSI pain symptoms were both significantly higher in the treatment group than in the control (P < 0.01). Within the treatment group, the score on NIH-CPSI pain symptoms was even more significantly improved in patients with the first attacks than in those already treated by other means (P < 0.01). No adverse effects were observed in either of the groups.
CONCLUSIONBoth the combined therapy of medication with local hyperthermia and simple local hyperthermia are effective, safe and tolerable in the treatment of CP pain symptoms, and the former is even more desirable, particularly for those with the first attacks of the symptoms.
Administration, Oral ; Adolescent ; Adrenergic alpha-Antagonists ; administration & dosage ; therapeutic use ; Adult ; Chronic Disease ; Combined Modality Therapy ; Doxazosin ; administration & dosage ; therapeutic use ; Drug Therapy ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Hyperthermia, Induced ; methods ; Male ; Middle Aged ; Pain ; etiology ; Pain Management ; Pain Measurement ; Prostatitis ; complications ; therapy ; Treatment Outcome ; Young Adult
3.De Novo and Inherited SETD1A Variants in Early-onset Epilepsy.
Xiuya YU ; Lin YANG ; Jin LI ; Wanxing LI ; Dongzhi LI ; Ran WANG ; Kai WU ; Wenhao CHEN ; Yi ZHANG ; Zilong QIU ; Wenhao ZHOU
Neuroscience Bulletin 2019;35(6):1045-1057
Early-onset epilepsy is a neurological abnormality in childhood, and it is especially common in the first 2 years after birth. Seizures in early life mostly result from structural or metabolic disorders in the brain, and the genetic causes of idiopathic seizures have been extensively investigated. In this study, we identified four missense mutations in the SETD1A gene (SET domain-containing 1A, histone lysine methyltransferase): three de novo mutations in three individuals and one inherited mutation in a four-generation family. Whole-exome sequencing indicated that all four of these mutations were responsible for the seizures. Mutations of SETD1A have been implicated in schizophrenia and developmental disorders, so we examined the role of the four mutations (R913C, Q269R, G1369R, and R1392H) in neural development. We found that their expression in mouse primary cortical neurons affected excitatory synapse development. Moreover, expression of the R913C mutation also affected the migration of cortical neurons in the mouse brain. We further identified two common genes (Neurl4 and Usp39) affected by mutations of SETD1A. These results suggested that the mutations of SETD1A play a fundamental role in abnormal synaptic function and the development of neurons, so they may be pathogenic factors for neurodevelopmental disorders.