Effect of electroacupuncture of "Zusanli" (ST36) combined with capeOX on apoptosis and ferroptosis in nude mice with colorectal cancer. / 电针"足三里"联合capeOXæ–¹æ¡ˆå¯¹ç»“ç›´è‚ ç™Œè£¸é¼ è‚¿ç˜¤ç»†èƒžå‡‹äº¡å’Œé“æ­»äº¡çš„å½±å“.

OBJECTIVES: To investigate the impact of combined treatment of colorectal cancer (CRC) with electroacupuncture (EA) and capeOX (combined administration of fluorouracil, oxaliplatin and capecitabine) on the tumor volume, weight, spleen coefficient, apoptosis and ferroptosis of tumor tissue, and liver and kidney functions in nude mice with CRC, so as to explore its mechanisms underlying inhibiting CRC and alleviating toxic reactions of capeOX. METHODS: Female Balb/c nude mice were randomly assigned to 3 groups：model, capeOX, and EA+capeOX, with 8 nude mice in each group. The CRC model was established by subcutaneous injection of colon cancer cells at the right inguinal region. Nude mice of the capeOX group received intraperitoneal injection of oxaliplatin for 1 day and gavage of capecitabine from day 2 to day 7. EA (1 mA, 2 Hz/100 Hz) was applied to bilateral "Zusanli" (ST36) for 20 min, once daily for 7 days. During the interven-tion, the tumor volume and weight were measured every day, and at the end of intervention, the weight of the tumor tissue and spleen were measured, with tumor volume difference and spleen coefficient calculated. The proportion of apoptotic cells was measured by flow cytometry, and the contents of serum malondialdehyde (MDA), alanine aninotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr) were detected using ELISA. The expression level of glutathione peroxidase 4 (GPX4, a key regulator for ferroptosis) protein of the tumor tissue was determined using Western blot. RESULTS: Compared to the model group, both the capeOX group and EA+capeOX group showed a decrease in the tumor volume (on day 3 and 4 in the capeOX group, and from day 2 to 7 in the EA+capeOX group) and body weight (P<0.05, on day 3 to 7 in the EA+capeOX group and on day 2 to 7 in the capeOX group), being evidently lower in the tumor volume on day 7 in the EA+capeOX than in the capeOX group (P<0.05), and evidently higher in the body weight on day 6 and 7 in the EA+capeOX group than in the capeOX group (P<0.05). In comparison with the model group, the tumor volume difference, tumor weight and spleen coefficient in both capeOX and EA+capeOX groups were significantly decreased (P<0.05), and MDA content in EA+capeOX group was significantly decreased (P<0.05), while the contents of ALT, BUN and Cr in the capeOX group, the proportion of apoptotic cells in both capeOX and EA+capeOX groups, and the GPX4 expression level in the EA+capeOX group were all significantly increased (P<0.05). The tumor volume difference, tumor weight, and contents of MDA, ALT, AST, BUN and Cr in the EA+capeOX group were markedly lower than in the capeOX group (P<0.05), while the spleen coefficient, proportion of apoptotic cells and GPX4 expression level in the EA+capeOX group were markedly higher than those in the capeOX group (P<0.05). CONCLUSIONS: EA of ST36 can enhance the effect of capeOX in inhibiting colorectal cancer growth in nude mice with CRC, which may be related with its functions in promoting tumor cell apoptosis, inhibiting ferroptosis, and modulating immune tolerance. In addition, EA can lower the side effects of capeOX in hematopoietic and immune, liver, and kidney functions.

Dynamic regulation of phosphorylation of NMDA receptor GluN2B subunit tyrosine residues mediates ketamine rapid antidepressant effects.

The rapid antidepressant effects of ketamine depend on the N-methyl-D-aspartate (NMDA) receptor containing 2B subunit (NR2B), whose function is influenced by its phosphorylated regulation and distribution within and outside synapses. It remains unclear if ketamine's rapid onset of antidepressant effects relies on the dynamic phosphorylated regulation of NR2B within and outside synapses. Here, we show that ketamine rapidlyalleviated depression-like behaviors and normalized abnormal expression of pTyr1472NR2B and striatal-enriched protein tyrosine phosphatase (STEP) 61 within and outside synapses in the medial prefrontal cortex (mPFC) induced by chronic unpredictable stress (CUS) and conditional knockdown of STEP 61, a key phosphatase of NR2B, within 1 hour after administration Together, our results delineate the rapid initiation of ketamine's antidepressant effects results from the restoration of NR2B phosphorylation homeostasis within and outside synapses. The dynamic regulation of phosphorylation of NR2B provides a new perspective for developing new antidepressant strategies.

[Short-term effectiveness of transverse antecubital incision for failed closed reduction of Gartland type â ¢ supracondylar humerus fractures in children].

Objective: To investigate the short-term effectiveness of transverse antecubital incision in the treatment of failed closed reduction of Gartland type Ⅲ supracondylar humeral fractures (SHFs) in children. Methods: Between July 2020 and April 2022, 20 children with Gartland type Ⅲ SHFs who failed in closed reduction were treated with internal and external condylar crossing Kirschner wire fixation through transverse antecubital incision. There were 9 boys and 11 girls with an average age of 3.1 years (range, 1.1-6.0 years). The causes of injuries were fall in 12 cases and fall from height in 8 cases. The time from admission to operation ranged from 7 to 18 hours, with an average of 12.4 hours. The healing of the incision and the occurrence of complications such as nerve injury and cubitus varus were observed after operation; the elbow flexion and extension range of motion after removing the gypsum, after removing the Kirschner wire, and at last follow-up were recorded and compared, as well as the elbow flexion and extension and forearm rotation range of motion at last follow-up between healthy and affected sides; the Baumann angle was measured on the X-ray film, and the fracture healing was observed. At last follow-up, the effectiveness was evaluated according to the Flynn elbow function evaluation criteria. Results: All incisions healed by first intention, and there was no skin necrosis, scar contracture, ulnar nerve injury, and cubitus varus. Postoperative pain occurred in the radial-dorsal thumb in 2 cases. The gypsum was removed and elbow flexion and extension exercises were started at 2-4 weeks (mean, 2.7 weeks) after operation, and the Kirschner wire was removed at 4-5 weeks (mean, 4.3 weeks). All the 20 patients were followed up 6-16 months, with an average of 12.4 months. The fracture healing time was 4-5 weeks, with an average of 4.5 weeks, and there was no complication such as delayed healing and myositis ossificans. The flexion and extension range of motion of the elbow joint gradually improved after operation, and there were significant differences between the time after removing the gypsum, after removing the Kirschner wire, and at last follow-up ( P<0.017). There was no significant difference in the flexion and extension of the elbow joint and the forearm rotation range of motion between the healthy and affected sides at last follow-up ( P>0.05). There was no significant difference in Baumann angle between the time of immediate after operation, after removing the Kirschner wire, and at last follow-up ( P>0.05). According to Flynn elbow function evaluation standard, 16 cases were excellent and 4 cases were good, the excellent and good rate was 100%. Conclusion: The treatment of Gartland type Ⅲ SHFs in children with failed closed reduction by internal and external condylar crossing Kirschner wire fixation through transverse antecubital incision has the advantages of complete soft tissue hinge behind the fracture for easy reduction and wire fixation, small incision, less complications, fast fracture healing, early functional recovery, reliable reduction and fixation, and can obtain satisfactory results.

Quantitative magnetic resonance spectroscopy of depression: The value of short-term metabolite changes in predicting treatment response.

Background: Although various prediction models of the antidepressant response have been established, the results have not been effectively applied to heterogeneous depression populations, which has seriously limited their clinical value. This study tried to build a more specific and stable model to predict treatment response in depression based on short-term changes in hippocampal metabolites. Materials and methods: Seventy-four major depressive disorder (MDD) patients and 20 healthy controls in the test set were prospectively collected and retrospectively analyzed. Subjects underwent magnetic resonance spectroscopy (MRS) once a week during 6 weeks of treatment. Hippocampal regions of interest (ROIs) were extracted by using a voxel iteration scheme combined with standard brain templates. The short-term differences in hippocampal metabolites between and within groups were screened. Then, the association between hippocampal metabolite changes and clinical response was analyzed, and a prediction model based on logistic regression was constructed. In addition, a validation set (n = 60) was collected from another medical center to validate the predictive abilities. Results: After 2-3 weeks of antidepressant treatment, the differences in indicators (tCho wee0-2, tCho wee0-3 and NAA week0-3) were successfully screened. Then, the predictive abilities of these three indicators were revealed in the logistic regression model, and the optimal prediction effect was found in d(tCho) week0-3-d(NAA) week0-3 (AUC = 0.841, 95%CI = 0.736-0.946). In addition, their predictive abilities were further confirmed with the validation set. Limitations: The small sample size and the need for multiple follow-ups limited the statistical ability to detect other findings. Conclusion: The predictive model in this study presented accurate prediction and strong verification effects, which may provide early guidance for adjusting the treatment regimens of depression and serve as a checkpoint at which the eventual treatment outcome can be predicted.

mPFC-rTMS for patients with insomnia disorder using resting-state functional magnetic resonance imaging: a protocol for a randomized controlled trial.

BACKGROUND: Insomnia is the most common sleep disorder. Repetitive transcranial magnetic stimulation (rTMS) is safe and effective for insomnia disorder (ID). Convergent evidence show that the medial prefrontal cortex (mPFC) may be involved in the regulation of sleep and awakening at the cortical level and may serve as a potential target of rTMS in the treatment of ID. The purpose of this clinical trial is to study the efficacy of mPFC-rTMS in the treatment ID and explore the neural mechanism using resting-state functional magnetic resonance imaging (fMRI). METHODS AND DESIGN: This will be a parallel-group randomized, patient- and assessor-blinded trial. The study will recruit 60 ID patients assigned to a real mPFC-rTMS group or a sham mPFC-rTMS group. The allocation ratio is 1:1, with 30 subjects in each group. Interventions will be administered five times per week over a 4-week period, with an 8-week follow-up period. All participants will undergo neuropsychological and fMRI evaluations. The primary outcome measure of this study is the change scores of the Pittsburgh Sleep Quality Index (PSQI). The secondary outcome measures include the fMRI measurements, the Hamilton Depression Scale (HAMD), the Hamilton Anxiety Scale (HAMA), a sleep diary, and a polysomnography. Assessment of all parameters will be performed at baseline, post-treatment, and during follow-up. DISCUSSION: It is expected that the study results will provide strong evidence of the effectiveness and the neural mechanism by which mPFC-rTMS improves sleep quality in ID patients. TRIAL REGISTRATION: Chinese Clinical Trials Register ChiCTR2100054154. Registered on 10 December 2021.

Interleukin-6-white matter network differences explained the susceptibility to depression after stressful life events.

BACKGROUND: Stressful life events (SLEs) are well-established proximal predictors of the onset of depression. However, the fundamental causes of interindividual differences in depression outcomes are poorly understood. This study addressed this depression susceptibility mechanism using a well-powered sample of adults living in China. METHODS: Healthy participants with SLEs (n = 185; mean = 47.51 years, 49.73% female), drawn from a longitudinal study on the development of depression, underwent diffusion tensor imaging, interleukin-6 (IL-6) level measurement, and trimonthly standardized clinical and scale evaluations within a two-year period. RESULTS: Receiver operating characteristic analyses indicated that reduced feeder connection and HIP.R nodal efficiency improved the predictive accuracy of post-SLEs depression (ORfeeder = 0.623, AUC = 0.869, P < 0.001; ORHIP = 0.459, AUC = 0.855, P < 0.001). The successfully established path analysis model confirmed the significant partial effect of SLEs-IL-6-white matter (WM) network differences-depression (onset and severity) (x2/8 = 1.453, goodness-of-fit [GFI] = 0.935, standard root-mean-square error of approximation [SRMR] = 0.024). Females, individuals with lower exercise frequency (EF) or annual household income (AHI) were more likely to have higher IL-6 level after SLEs (ßint-female⁎SLEs = -0.420, P < 0.001; ßint-exercise⁎SLEs = -0.412, P < 0.001; ßint-income⁎SLEs = -0.302, P = 0.005). LIMITATIONS: The sample size was restricted due to the limited incidence rate and prospective follow-up design. CONCLUSIONS: Our results suggested that among healthy adults after SLEs, those who exhibited abnormal IL-6-WM differences were susceptible to developing depression. Females, lower AHI or EF might account for an increased risk of developing these abnormal IL-6-WM differences.

A new scheme of global feature management improved the performance and stability of radiomics model: a study based on CT images of acute brainstem infarction.

OBJECTIVE: The performance and stability of radiomics model caused by dimension reduction remain being confronted with major challenges. In this study, we aimed to propose a new scheme of global feature management independent of dimension reduction to improve it. METHODS: The non-contrast computed tomography (NCCT) images of acute brainstem infarction (ABI) from two medical centers were used as test and validation sets. A new scheme was constructed based on global feature management, and the traditional scheme dependent on dimension reduction was used as control. The radiomic features of NCCT images were extracted in Matlab R2013a. The performance of prediction model was evaluated by the generalized linear model (GLM) and multivariate logistic regression. And, the stability of radiomics model was evaluated with the difference of area under curve (AUC) between the test and validation sets. RESULTS: Compared with the traditional scheme, the new scheme presented a similar detection performance (AUC: 0.875 vs. 0.883), yet a better performance in predicting prognosis (AUC: 0.864, OR = 0.917, p = 0.021 vs. AUC:0.806, OR = 0.972, p = 0.007). All these results were well verified in an independent validation set. Moreover, the new scheme showed stronger stability in both the detection model (ΔAUC: 0.013 vs. 0.039) and prediction model (ΔAUC = 0.004 vs. 0.044). CONCLUSION: Although there might be several limitations, this study proved that the scheme of global feature management independent of dimension reduction could be a powerful supplement to the radiomics methodology. KEY POINTS: • The new scheme (Swavelet) presented similar detection performances for ABI with the traditional scheme. • A better predictive performance for END was found in the new scheme (Swavelet) compared with the traditional scheme. • Stronger model stability was found in both the detection and prediction models based on the new scheme.

Mediation on the Association Between Stressful Life Events and Depression by Abnormal White Matter Microstructures.

BACKGROUND: Stressful life events (SLEs) are an important causal factor in depression; however, the mechanism by which SLEs cause depression remains unclear. Recent studies suggested that white matter (WM) microstructures might be a potential mediator between SLEs and depression. Hence, we aimed to investigate the concrete correspondence among them using mediation effect models. METHODS: In participants (N = 194) with SLEs experience prospectively recruited from six residential communities, WM microstructures were detected with diffusion tensor imaging. The interrelationship among SLEs, WM microstructures, and depression was explored with multiple linear regression models and logistic regression models. Furthermore, the influence of WM microstructures on the association between SLEs and depression was tested with mediation effect models. RESULTS: Successfully established mediation effect models showed the specific influence of fractional anisotropy of the corpus callosum and left uncinate fasciculus on the association between SLEs and depression onset (ab path = 0.032; ab path = 0.026, respectively) and between SLEs and depressive severity (ab path = 0.052; ab path = 0.067, respectively). In addition, significant total mediation effects on the association between SLEs and depression onset (ab path = 0.031) and severity (ab path = 0.075) through fractional anisotropy of the corpus callosum and left uncinate fasciculus were noted. CONCLUSIONS: WM microstructure alterations impose a substantial mediation effect on the association between SLEs and depression, which suggest that changes in WM microstructure integrity might increase the risk of depression onset and unfavorable disease courses induced by the SLEs.

Polymorphisms in MicroRNA Genes Associated with Schizophrenia Susceptibility but Not with Effectiveness of MECT.

Schizophrenia (SCZ) is a common and complex psychiatric disease associated with hereditary and environmental risk factors. MicroRNAs (miRNAs or miRs) are small, noncoding RNA molecules that endogenously regulate gene expression. Single nucleotide polymorphisms (SNPs) in related miRNA genes are associated with susceptibility of the disorder. We wonder if the SNPs have influence on the effectiveness of modified electroconvulsive therapy (MECT) for SCZ. rs1625579 within miR-137, rs6577555 within miR-34, and rs2296616 within miR-107 were sequenced in 150 cases and 150 controls to check the potential association between the SNPs and SCZ. Our results showed that allele G in rs1625579 (p = 0.005, adjusted OR = 1.379, 95%CI = 1.108 - 1.634), allele A in rs6577555 (p = 0.014, adjusted OR = 1.246, 95%CI = 1.045 - 1.463), allele G in rs2296616 (p < 0.001, adjusted OR = 1.646, 95%CI = 1.374 - 1.879) are positively associated with the disorder risk. MECT courses did significantly decrease the level of the miRNAs, except for the variant of rs2296616 with the AA genotype. Schizophrenic phenotypes assessed by the positive and negative syndrome scale (PANSS) were improved after MECT, and there was no significant relevance observed between the effectiveness of MECT and the variants of these loci. Thus, our findings indicate that polymorphisms within the loci may be involved in the pathogenesis of SCZ, and MECT is effective and unbiased for patients harboring different genotypes of the loci.

Enhancing excitatory projections from the ventral subiculum to the nucleus accumbens shell contribute to the MK-801-induced impairment of prepulse inhibition.

Prepulse inhibition (PPI), a measure of sensorimotor gating, has been shown to be disrupted in several animal models of neuropsychiatric disorders, such as schizophrenia. The neural circuits involving the hippocampus and nucleus accumbens (NAC) have been studied in rats to uncover the neurochemical and neuroanatomical substrates that regulate PPI. Majority of the studies of the hippocampus on PPI to date have been focused on CA1, CA2, and dentate gyrus (DG) area. Little is known about the role of the subiculum, which maintains the hippocampal formation intact, on the sensorimotor gating. In this study, the PPI disruption was induced by intraperitoneal injection of MK-801 in rats, and the neuronal activity in the dorsal and ventral subiculum by c-Fos immunostaining was examined. The projections from the subiculum to the nucleus accumbens (NAC) were detected by retrograde tracing of cholera toxin B subunit, in the PPI dysfunctional animals. The results showed an increase in neuronal activity in the ventral subiculum (vSub) while remaining constant in the dorsal subiculum during PPI disruption. The excitatory projections from the vSub to the NAC shell were significantly enhanced when PPI was disrupted. Muscimol Inhibition of vSub could significantly ameliorate the MK801-induced PPI deficit. This data suggests that the enhancement of neuronal activity in the vSub was associated with the PPI impairment, possibly due to the enhanced excitatory output from vSub the NAC shell.

Functional Status of Hypothalamic-Pituitary-Thyroid and Hypothalamic-Pituitary-Adrenal Axes in Hospitalized Schizophrenics in Shanghai.

OBJECTIVE: Neuroendocrine dysfunction is related to the pathogenesis of mental disorders, but conclusions from clinical research lack consistency. We aimed to investigate the neuroendocrinal pathophysiology and its correlation with clinical symptoms in patients with schizophrenia. METHODS: The present cross-sectional study included 486 inpatients with schizophrenia admitted at a psychiatric hospital in Shanghai within one year, and 154 healthy controls (HC) matched on age and gender. The serum hemoconcentrations of thyroid-stimulating hormone (TSH), total triiodothyronine (TT3), total thyroxine (TT4), free triiodothyronine (FT3), free thyroxine (FT4), adrenocorticotrophic hormone (ACTH), and cortisol (COR) were measured via electrochemical luminescence immunoassay. Pathophysiological conversions of neuroendocrine were then associated with gender, age, age at onset, antipsychotic treatment using hierarchical multiple linear regression. RESULTS: When compared to HC, the schizophrenia group showed elevated ACTH and COR levels and decreased TT3 and TT4 levels (p's < 0.05). First-episode patients showed lower TSH and higher FT3 and FT4 (p's < 0.05) compared to recurrent patients. Female patients showed higher TSH and lower TT3, FT3, and ACTH levels (p's < 0.05) compared to males. We observed the area under the curve (AUC) of the predictive model to distinguish between schizophrenia and HC to be 0.737 among total samples and between first-episode and recurrent schizophrenia to be 0.890 among subgroups. CONCLUSIONS: Decreased TT3 and TT4 and elevated ACTH and COR levels appear to be associated with schizophrenia symptoms. The chronic recurrent trait of schizophrenia may cause long-term effects on FT3 and FT4 while changes in thyroid, and adrenal function as a result of mental disorder varied with gender. The pathophysiological parameters provide fair to good accuracy of these models.

[Response inhibition and emotional responding in attention-deficit/hyperactivity disorder with comorbid disruptive, impulse-control, and conduct disorders].

OBJECTIVE: To characterize the traits of neuropsychological functioning deficits in patients with attention-deficit/ hyperactivity disorder (ADHD) with comorbid disruptive, impulse-control, and conduct disorders (DICCD). METHODS: Twenty out-patients with ADHD, 20 with ADHD with comorbid DICCD, and 20 with DICCD, all aged 6-16 years, were enrolled in this study, with 20 healthy subjects matched for age, gender and IQ serving as the healthy controls. The patients were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, Fifth Revision (DSM-5). All the subjects were assessed with Golden Stroop test and emotional Stroop test to evaluate their response inhibition and emotional responding. RESULTS: In Golden Stroop test, the interference scores (IGs) of errors and reaction time both differed significantly among the groups (P < 0.05), and were the highest in patients with ADHD only. In emotional Stroop test, the mean reaction time (MRT) showed significant differences among the groups (P < 0.05); the MRT of positive- congruent trials in ADHD with comorbid DICCD group was shorter than that in ADHD group but longer than that in group DICCD; the MRT in the 3 case groups were all longer than that in the control group. The MRT of both positive-incongruent trials and negative-congruent trials in ADHD with comorbid DICCD group and DICCD group was shorter than that in ADHD group but longer than that in the control group. The MRT of negative- incongruent trials in DICCD group was shorter than that in ADHD group and ADHD with comorbid DICCD group but longer than that in the control group. CONCLUSIONS: The response inhibition deficit and abnormal emotional responding are the core symptoms of ADHD. Bias emotional stimuli may render response inhibitory dysfunction in patients with DICCD with callous-unemotional traits of emotional responding disorder, especially in dealing with negative emotional trials, while the comorbidity of ADHD and DICCD tends to have the emotional response trait of DICCD.

Hyperactivity and disruption of prepulse inhibition induced by NMDA infusion of the rat ventral hippocampus: Comparison of uni- and bilateral stimulation.

Numerous studies have demonstrated that NMDA stimulation of the rat ventral hippocampus induces hyperactivity and disruption of prepulse inhibition (PPI) and some reports have provided insights into the different neural substrates of these two effects. Bilateral or unilateral NMDA stimulation have been applied in these studies, but have never been compared directly. Thus, results of these studies can be related to each other only with reservations. Therefore - given the substantial interest in the behavioral effects of hippocampal overactivity and the neural substrates underlying PPI with respect to schizophrenia - the present experiments compared hyperactivity and PPI disruption following uni- or bilateral NMDA stimulation of the ventral hippocampus. Both bilateral and unilateral infusion of NMDA (0.5µg/0.5µl/side) induced hyperactivity in the open field and disrupted PPI. The major difference noted between unilateral and bilateral infusions was that the effects of bilateral infusions were stronger than those of the unilateral infusions.

[Application of pedicle reducing tension flap in the distally-based pedicle flap operation].

OBJECTIVE: To explore the effectiveness of reducing tension method on the survival and appearance of distally-based pedicle flap. METHODS: Between October 2009 and February 2011, 27 cases of defect of extremity skin and soft tissue were repaired with distally-based pedicle flap through reducing tension flap method. There were 19 males and 8 females with an average age of 31.5 years (range, 17-58 years). Defects were caused by traffic accident in 14 cases, by machine in 9 cases, by surgical infection in 3 cases, and by tumor excision in 1 case. The locations were the distal tibia in 7 cases, the ankle-joint in 12 cases, the foot in 5 cases, the palm in 2 cases, and the dorsum of hand in 1 case. The time from injury to hospitalization was 1-19 hours with an average of 10 hours. The size of defect ranged from 5.2 cm x 3.8 cm to 14.0 cm x 5.8 cm. The size of distally-based pedicle flap ranged from 5.5 cm x 4.5 cm to 14.5 cm x 6.5 cm. The donor sites were sutured directly in 6 cases and were repaired with skin grafting in 21 cases. RESULTS: All reducing tension flaps survived. Partial necrosis occurred in the distally-based pedicle flap in 3 cases at 7 days after operation, which was cured after dressing change and skin grafting in 1 case, after excision of necrosis skin edge and direct suture in 2 cases. The other flaps survived and wounds achieved primary healing. The incisions at donor sites healed by first intention and skin grafting survived. Twenty-six cases were followed up 6-12 months (mean, 7.5 months). The appearance and texture of the flaps were good. CONCLUSION: Pedicle reducing tension flap could promote the survival and the appearance of distally-based pedicle flap.

[Screening of differential expression genes of human skin epidermal stem cells at different development stages by cDNA microarray technique].

OBJECTIVE: To analyze expression characteristics of human skin epidermal stem cell at different developmental stages, and to explore its biological significance. METHODS: Health skin samples from 28-32 w fetuses (F group), 4-12 y children (C group), and 35-55 y adult (A group) were harvested, with 10 cases in each group. Epidermis were separated using trypsin digestion and EDTA, and human epidermal stem cells were isolated and purified with type IV collagen attachment method. The monoclonal antibody of integrin beta1 and keratin 19 were used for detection and identification of epidermal stem cells by immunohistochemical staining. Total RNA was extracted from above cells by Trizol one-step method, and were detected by formaldehyde denaturing agarose gel electrophoresis. Probes were prepared and hybridized into cDNA microarray for scanning fluorescent signals and analysis of images, with two-fold differential expression value for screening. Significantly up/down-regulated genes were selected for verification by real time RT-PCR. RESULTS: By comparing expression profile between A and C groups, a total of 1808 genes with differential expression were detected, including 1089 up-regulated genes and 719 down-regulated genes, and they were classified into 128 categories. Among them, 1462 genes were known (found in GeneBank), 346 genes were unknown. A total of 4534 genes with differential expression were detected between C and F groups, in which 1783 genes were up-regulated and 2751 genes were down-regulated, and they were classified into 216 categories. Among them, 3577 genes were known (found in GeneBank), and 957 genes were unknown. There were 1104 genes with differential expression consistently detected in F, C and A groups, which were classified into 32 categories according to gene function. Among them, 94 genes were consistently up-regulated and 75 genes consistently down-regulated. Test results of real time RT-PCR were in accordance with above-mentioned results. CONCLUSIONS: Gene expression profiles of epidermal stem cells cultured in vitro, harvested from fetuses, children, and adult, exhibit obvious difference. This may be closely related to different stages of proliferation and differentiation of human epidermal stem cell and self-repair ability of wound at different developmental stages.