Directed differentiation of human induced pluripotent stem cells into midbrain / 南方医科大学学报

OBJECTIVE@#To establish an efficient protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into functional midbrain dopaminergic progenitor cells (DAPs) in vitro.@*METHODS@#hiPSCs were induced to differentiate into DAPs in two developmental stages. In the first stage (the first 13 days), hiPSCs were induced into intermediate cells morphologically similar to primitive neuroepithelial cells (NECs) in neural induction medium containing a combination of small molecule compounds. In the second stage, the intermediate cells were further induced in neural differentiation medium until day 28 to obtain DAPs. After CM-DiI staining, the induced DAPs were stereotactically transplanted into the right medial forebrain bundle (MFB) of rat models of Parkinson's disease (PD). Eight weeks after transplantation, the motor behaviors of PD rats was evaluated. Immunofluorescence assay of brain sections of the rats was performed at 2 weeks after transplantation to observe the survival, migration and differentiation of the transplanted cells in the host brain microenvironment.@*RESULTS@#hiPSCs passaged stably on Matrigel showed a normal diploid karyotype, expressed the pluripotency markers OCT4, SOX2, and Nanog, and were positive for alkaline phosphatase. The primitive neuroepithelial cells obtained on day 13 formed dense cell colonies in the form of neural rosettes and expressed the neuroepithelial markers (SOX2, Nestin, and PAX6, 91.3%-92.8%). The DAPs on day 28 highly expressed the specific markers (TH, FOXA2, LMX1A and NURR1, 93.3-96.7%). In rat models of PD, the hiPSCs-DAPs survived and differentiated into TH+, FOXA2+ and Tuj1+ neurons at 2 weeks after transplantation. Eight weeks after transplantation, the motor function of PD rats was significantly improved as shown by water maze test (P < 0.0001) and apomorphine-induced rotation test (P < 0.0001) compared with rats receiving vehicle injection.@*CONCLUSION@#HiPSCs can be effectively induced to differentiate into DAPs capable of differentiating into functional neurons both in vivo and in vitro. In rat models of PD, the transplanted hiPSCs-DAPs can survive for more than 8 weeks in the MFB and differentiate into multiple functional neurocytes to ameliorate neurological deficits of the rats, suggesting the potential value of hiPSCs-DAPs transplantation for treatment of neurological diseases.

Clinical analysis of inflatable video-assisted mediastinoscopic transhiatal esophagectomy combined with laparoscopy / 中华外科杂志

Objective: To examine the safety and effectiveness of inflatable video-assisted mediastinoscopic transhiatal esophagectomy (IVMTE). Methods: Totally 269 patients admitted to the Anhui Provincial Hospital of Anhui Medical University who underwent IVMTE (IVMTE group, n=47) or thoracoscopy combined with minimally invasive Mckeown esophageal cancer resection (MIME group, n=222) from September 2017 to December 2021 were analyzed retrospectively. There were 31 males and 16 females in IVMTE group, aged (68.6±7.5) years (range: 54 to 87 years). There were 159 males and 63 females in MIME group, aged (66.8±8.8) years (range: 42 to 93 years). A 1∶1 match was performed on both groups by propensity score matching, with 38 cases in each group. The intraoperative conditions and postoperative complication rates of the two groups were compared by t test, Wilcoxon rank, χ2 test, or Fisher exact probability method. Results: Patients in IVMTE group had less intraoperative bleeding ((96.0±39.2) ml vs. (123.8±49.3) ml, t=-2.627, P=0.011), shorter operation time ((239.1±47.3) minutes vs. (264.2±57.2) minutes, t=-2.086, P=0.040), and less drainage 3 days after surgery (85(89) ml vs. 675(573) ml, Z=-7.575, P<0.01) compared with that of MIME group. There were no statistically significant differences between the two groups in terms of drainage tube-belt time, postoperative hospital stay, and lymph node dissection stations and numbers (all P>0.05). The incidence of Clavien-Dindo grade 1 to 2 pulmonary infection (7.9%(3/38) vs. 31.6%(12/38), χ²=6.728, P=0.009), total complications (21.1%(8/38) vs. 47.4%(18/38), χ²=5.846, P=0.016) and total lung complications (13.2%(5/38) vs. 42.1%(16/38), χ²=7.962, P=0.005) in the IVMTE group were significantly lower. Conclusion: Inflatable video-assisted mediastinoscopic transhiatal esophagectomy combined with laparoscopic esophagectomy is safe and feasible, which can reach the same range of oncology as thoracoscopic surgery.

Effects of ring finger and tryptophan-aspartic acid 2 on dendritic spines and synapse formation in cerebral cortex neurons of mice / 南方医科大学学报

OBJECTIVE@#To clarify the functional effects of differential expression of ring finger and tryptophan-aspartic acid 2 (RFWD2) on dendritic development and formation of dendritic spines in cerebral cortex neurons of mice.@*METHODS@#Immunofluorescent staining was used to identify the location and global expression profile of RFWD2 in mouse brain and determine the co-localization of RFWD2 with the synaptic proteins in the cortical neurons. We also examined the effects of RFWD2 over-expression (RFWD2-Myc) and RFWD2 knockdown (RFWD2-shRNA) on dendritic development, dendritic spine formation and synaptic function in cultured cortical neurons.@*RESULTS@#RFWD2 is highly expressed in the cerebral cortex and hippocampus of mice, and its expression level was positively correlated with the development of cerebral cortex neurons and dendrites. RFWD2 expression was detected on the presynaptic membrane and postsynaptic membrane of the neurons, and its expression levels were positively correlated with the length, number of branches and complexity of the dendrites. In cultured cortical neurons, RFWD2 overexpression significantly lowered the expressions of the synaptic proteins synaptophysin (P < 0.01) and postsynapic density protein 95 (P < 0.01), while RFWD2 knockdown significantly increased their expressions (both P < 0.05). Compared with the control and RFWD2-overexpressing cells, the neurons with RFWD2 knockdown showed significantly reduced number of dendritic spines (both P < 0.05).@*CONCLUSION@#RFWD2 can regulate the expression of the synaptic proteins, the development of the dendrites, the formation of the dendritic spines and synaptic function in mouse cerebral cortex neurons through ubiquitination of Pea3 family members and c-Jun, which may serve as potential treatment targets for neurological diseases.

Review of 3D Printing Technology in Development and Application of Innovative Chinese Medicines / 中国实验方剂学杂志

As an effective additive manufacturing technology, 3D printing technology has been widely used in aerospace, construction, machinery manufacturing and other fields, but it is rarely used in the pharmaceutical field, especially in the field of traditional Chinese medicine (TCM). In 2015, Spritam instant tablets produced by 3D printing technology greatly promoted the development and research of 3D printing technology in the pharmaceutical industry. In this paper, we reviewed three types of 3D printers (binder jet, material extrusion and inkjet printing) with high compatibility in the pharmaceutical industry, including the working principles, printing materials, product characteristics and others. In addition, the application prospects and existing problems of 3D printing technology in the production of innovative preparations of TCM, the development of innovative medical devices and the development of innovative functional foods were analyzed. The paper aimed to provide a reference for the development and preparation of innovative TCM by 3D printing technology.

Contralateral head leading turning accompanied by ipsilateral eye staring in a patient with seizure onset from posterior inferior temporal sulcus, a SEEG case report

The epileptic eye and head movements during epileptic seizures may be much more complicated thanpeople originally understood, which can be ipsilateral or contralateral to the electroencephalographyfocus. Here, we describe a male patient with drug resistant focal seizures associated with a directionalseparation between head and eye movement before evolving into generalized tonic-clonic seizure.His contralateral head leading turning showed forced, sustained, and unnatural features companiedby ipsilateral eye staring. Stereoeletroencephalography monitoring was performed, and 4 habitualseizures were recorded over 5 days. Three seizures showed left head leading turning and generalizedtonic-clonic seizure, and only one showed dizziness and ringing in the ears. All the seizures showedthat the ictal onset contacts were located in the posterior inferior temporal sulcus which borders onthe anterolateral part of medial superior temporal area. The patient underwent a resection includingtemporooccipital region, and the histopathology showed focal cortical dysplasia type Ic. He has beenseizure free for two years after operation. The scores of the intelligence and memory quotient improvedhalf year after operation.

Optimization of two-phase aqueous extraction for Astragali Radix by central composite design-response surface method / 中成药

AIM To optimize the two-phase (ethanol-dipotassium phosphate) aqueous extraction for Astragali Radix by central composite design-response surface method.METHODS With ethanol concentration,dipotassium hydrogen phosphate solution concentration and Astragali Radix feed ratio as influencing factors,together with extraction rates of total flavonoids and total saponins as evaluation indices,the technology optimization was accomplished by central composite design-response surface method.RESULTS The optimal conditions were determined to be 27.97％ for ethanol concentration,22.03％ for dipotassium hydrogen phosphate solution concentration,and 1 ∶ 58.85 for Astragali Radix feed ratio.The extraction rate of total flavonoids reached 74.13％ (concentrated in the upper layer),while that of total saponins reached 81.34％ (concentrated in the lower layer).CONCLUSION With good predictability,this simple and accurate method can be used for the extraction of total flavonoids and total saponins in Astragali Radix.

Kuntai capsule combined with gonadotropin releasing hormone agonist in treatment of moderate-severe endometriosis: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Kuntai Capsule (KC), a Chinese patent medicine, in add-back therapy for gonadotropin-releasing hormone agonist (GnRH-a) treatment for moderate-severe endometriosis (EM).</p><p><b>METHODS</b>Totally 100 patients suffering from stage III/IV EM, who were confirmed by laparoscopic surgery were randomly assigned to the GnRH-a group (A) and the KC combined GnRH-a group (B), 50 in each group. Patients in Group A were hypodermically injected with goserelin (3.6 mg), once per 4 weeks. Those in Group B additionally took KC, 4 pills each time, three times per day. The therapeutic course for all was 12 weeks. Serum levels of estradiol (E2), follicle stimulating hormone (FSH), bone gamma-carboxyglutamic-acid-containing proteins (BGP) were measured respectively. Kupperman Menopausal Index (KMI) and bone mineral density (BMD) of the lumbar vertebra were also compared between the two groups.</p><p><b>RESULTS</b>Serum levels of E2 and FSH both significantly decreased in the two groups at week 12 of the treatment (P < 0.05), when compared with pre-treatment. Compared with before treatment in the same group, KMI increased in the two groups (P < 0.05). Compared with before treatment in the same group, BMI decreased in the two groups with no statistical difference (P > 0.05). Serum BGP increased after 12-week treatment (P < 0.05). Compared with Group A after treatment, serum levels of E2 and FSH both significantly increased in Group B (P < 0.05). There was no statistical difference in KMI between the two groups (P > 0.05). As for the incidence of menopausal symptoms, better effects in improving symptoms such as hot flashes, sleep disorders, and vaginal dryness were obtained in Group B than in Group A (P < 0.05). There was no significant difference in the post-pre-treatment difference of BMI between the two groups, but with statistical post-pre-treatment difference in the BGP level (P < 0.05).</p><p><b>CONCLUSIONS</b>HKC combined GnRH-a could effectively reduce GnRH-a treatment induced partial low estrogen symptoms, improve increased serum BGP levels after GnRH-a therapy.</p>

Video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in thoracic cavity: analysis of 60 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and safety of video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity.</p><p><b>METHODS</b>The clinical data of 120 patients who underwent esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity from March to December 2011 was analyzed retrospectively. In the video-assisted thoracoscopic surgery group, there were 60 patients [41 male and 19 female patients with aver age of (62 ± 7) years old] who underwent video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. In the routine thoracotomy group, there were 60 patients [39 male and 21 female patients with aver age of (62 ± 9) years old] who underwent routine thoracotomy esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. Operation time, intra-operative blood loss, postoperative total thoracic drainage in 3 days, total number of harvested lymph nodes, hospitalization, cost of hospitalization and complications were compared between the two groups.</p><p><b>RESULT</b>The operations were carried out successfully in two groups. There was no perioperative death in all patients. There was no statistical difference in intra-operative blood loss, postoperative total thoracic drainage and cost of hospitalization between the two groups. Operation time of rideo-assisted thoracoscopic surgery group was significantly longer than that of thoracotomy group ((188 ± 38) minutes vs. (138 ± 50) minutes, t = 6.171, P = 0.000), but postoperative hospitalization was significantly lower ((14 ± 3) d vs. (18 ± 6) d, t = -4.093, P = 0.000) and total number of harvested lymph nodes was lower (17 ± 9 vs. 21 ± 11, t = -2.058, P = 0.042). There was significantly statistical difference in total postoperative main complication (25.0% vs. 48.3%, χ(2) = 7.033, P = 0.008). And postoperative incisional infection of VATE group patients was significantly lower than that of thoracotomy group patients (6.7% vs. 25.0%, χ(2) = 7.566, P = 0.006).</p><p><b>CONCLUSIONS</b>Video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity is technically feasible and safe, with minimized trauma and quick recovery. The recent result is satisfactory.</p>

Study of aniline's penetration capability through rats' complete skin in vitro and normal lab gloves / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the permeability of intact mouse abdominal skin to aniline and the protective capability of two typical lab gloves against aniline.</p><p><b>METHODS</b>A Franz diffusion cell was used to perform in vitro transdermal absorption test and glove permeation test for aniline (0.102 mg/ml and 0.010 mg/ml). The permeabilities of intact mouse abdominal skin and gloves to aniline were measured by high performance liquid chromatography-diode array detection.</p><p><b>RESULTS</b>The transdermal penetration of the two concentrations of aniline followed zero order kinetics within 12 h, exhibiting total aniline permeabilities within 24 h of 51.71% and 48.31%, respectively. The absorption liquid had an aniline concentration of at least 18 µg/L. The medical disposable latex glove could not stop the penetration of 0.010 mg/ml aniline, but the industrial natural latex glove could.</p><p><b>CONCLUSION</b>The penetration of 0.102 mg/ml and 0.010 mg/ml aniline through the mouse abdominal skin follows zero order kinetics within 12 h. The medical disposable latex glove cannot stop the penetration of 0.010 mg/ml aniline, but the industrial natural latex glove can.</p>

Whole brain irradiation for non-small-cell lung cancer with brain metastasis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the time of whole brain irradiation and the prognostic factors for non-small lung cancer patients with brain metastasis.</p><p><b>METHODS</b>From August 1996 to December 2003, 147 patients with brain metastasis from non-small cell lung cancer received whole brain irradiation. The patients were divided into two groups: with or without symptoms caused by brain metastasis, each group was then divided into two sub-groups, early whole brain irradiation group (the interval between the diagnosis of brain metastasis and the brain irradiation < or = one month) and late group ( the interval > one month ). Univariate and multivariate analysis (Cox regression) as well as Kaplan-Meier method in SPSS software package 11.5 was used to analyze the data of the 147 patients including 72 with brain metastasis symptom and 75 without.</p><p><b>RESULTS</b>The median survival time (MS) of patients with or without extracranial metastasis was 9.9 months and 11.3 months (P = 0.0002). Multivariate analysis indicated that extracranial metastasis was an independent prognostic factor (P = 0.0004). For 72 patients with brain metastasis symptom, the MS of the patients with and without extracranial metastasis was 9.3 months and 11.3 months (P = 0.0036). The MS of patients with early and late whole brain irradiation was 11.4 months and 9.2 months (P = 0.001). Multivariate analysis showed that extracranial metastasis, the interval between the diagnosis of brain metastasis and the whole brain irradiation were independent prognostic factors. However, for 75 patients without brain metastasis symptom, the MS difference of those with early or late whole brain irradiation was not statistically significant (P = 0.1643).</p><p><b>CONCLUSION</b>The extracranial metastasis in non-small cell lung cancer patients with brain metastasis is an independent prognostic factors. Early whole brain irradiation may improve the survival for those with brain metastasis symptoms.</p>

Simulation and analysis of ethanol concentration response to enzyme amount changes in Saccharomyces cerevisiae glycolysis pathway model / 生物工程学报

Metabolome has become an important part of Systems Biology, and a large set of data has already gained by applying the methods of metabolome. How to deal with the data and how to combine data of metabolome with data of other omics are problems that can not be ignored. An Enzyme Amount Multiple Factor was imported into the enzyme kinetic equation. When the enzyme amount in the system changed, in silico model, it means to alter the Enzyme Amount Multiple Factor. In order to observe ethanol concentration response to enzyme amount changes in S. cerevisiae glycolysis pathway model, enzyme amount was separately set at high and low level, the corresponding Enzyme Amount Multiple Factor value was 10 and 0.1, relatively. Based on the result of simulation, twelve enzymes in pathway were separated into two classes, class I and class II by cluster analysis. The four enzymes belonging to class I, ADH, HK, PFK and PDC, all catalyze irreversible reactions. The six out of eight enzymes belonging to class II, ALD, GAPDH, GlcTrans, lpPEP, PGI and TIM, catalyze reversible reactions. The other two enzymes belonging to class II, lpGlyc and PK, catalyze irreversible reactions. Based on this method, data of metabolome and proteomics are easily integrated to accomplish relatively overall analysis of system properties.

Highly selective inhibition of inducible nitric oxide synthase in vitro by a tripeptide as a new arginine analog / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of a tripeptide, a new arginine analog, on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS).</p><p><b>METHODS</b>Macrophages challenged with lipopolysaccharide were cultured to test the inhibitory effect of the arginine analog of different concentrations on iNOS. Primarily cultured human umbilical vein endothelial cells (HUVECs) treated with insulin were used to test the effect of the analog on endothelial NOS (eNOS).</p><p><b>RESULTS</b>The new arginine analog significantly inhibited NO production in the macrophages in a dose-dependent manner, but had little effect on insulin-stimulated NO production in HUVECs. The new analog could significantly suppress iNOS activity, but had no significant inhibitory effect on constitutive NOS activity or eNOS activity. Western blot analysis showed that the new analog did not significantly affect the protein expression of iNOS.</p><p><b>CONCLUSION</b>The new analog is a NOS inhibitor with high selectivity on iNOS.</p>

Recombinant adenovirus-p53 gene therapy combined with radiotherapy for head and neck squamous-cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of recombinant adenovirus-p53 gene (Gendicine) therapy combined with radiotherapy for head and neck squamous-cell carcinoma (HNSCC).</p><p><b>METHODS</b>From Oct. 2001 to May 2003, a randomized controlled clinical trial on Gendicine combined with radiation in 36 patients (gene therapy + radiotherapy, GTRT) vs. radiotherapy alone in 33 patients (RT) with HNSCC was completed. In the GTRT group, Gendicine 1 x 10(12) VP (virus particle) was injected intratumorally once a week for eight weeks, and concurrently followed by irradiation. For both groups, the conventional fractionation 2 Gy/f, five fractions a week to a total dose of 70 Gy, was given to either primary tumor or neck lymph nodes. Tumor response was assessed by CT image at 40 Gy, 70 Gy, 2 months after treatment to evaluate the response rate of CR, PR, SD and PD.</p><p><b>RESULTS</b>Wild-type p53 gene significantly enhanced radiotherapeutic effectiveness in patients with HNSCC (P < 0.05). The CR rate of tumors treated by GTRT was increased by nearly 2.31 times more than that of tumors treated by RT alone. No dose-limiting toxicity and adverse events were noted, except transient fever after Gendicine administration.</p><p><b>CONCLUSION</b>Intratumoral injection of Gendicine to HNSCC patients is safe and effective. The apparent improved results of combined therapy with Gendicine and radiation suggest that p53 gene therapy has promising therapeutic potential in cancer treatment.</p>