Objective To study biological rule of recombinant human bone morphogenetic protein 2 (rhBMP-2) in regulating the expression of vascular endothelial growth factor (VEGF) of adipose-derived stem cells (ADSCs) at different induced concentrations and time points at gene level and protein level. Methods ADSCs were separated from adult human adipose tissues and cultured until passage 3. After ADSCs were induced by rhBMP-2 in concentrations of 0, 50, 100, and 200 ng/ mL respectively for 24 hours, and by 100 ng/mL rhBMP-2 for 3, 6, 12, 18, 24, 36, and 48 hours (ADSCs were not induced at corresponding time point as controls) respectively, the VEGF mRNA and protein expressions were detected by RT-PCR and ELISA. Results The VEGF mRNA and protein expressions induced by rhBMP-2 were concentration-dependent; the expressions were highest in a concentration of 100 ng/mL. The VEGF mRNA expression in concentrations of 50, 100, and 200 ng/mL were significantly higher than that in a concentration of 0 ng/mL (P lt; 0.05); and the expression in concentration of 100 ng/ mL was significantly higher than that in concentrations of 50 and 200 ng/mL (P lt; 0.05). The VEGF protein expression in a concentration of 100 ng/mL was significantly higher than that in the other concentrations (P lt; 0.05). The VEGF mRNA and protein expressions induced by rhBMP-2 were time-dependent. The VEGF mRNA and protein expressions at 3 and 6 hours after induction were significantly lower than those of non-induced ADSCs (P lt; 0.05); the expressions were lower at 12 hours after induction, showing no significant difference when compared with those of non-induced ADSCs (P gt; 0.05); the expressions reached peak at 18 and 24 hours after induction, and were significantly higher than those of non-induced ADSCs (P lt; 0.05); the expressions decreased in induced and non-induced ADSCs at 36 and 48 hours, showing no significant difference between induced and non-induced ADSCs (P gt; 0.05). Conclusion rhBMP-2 adjusts VEGF expression of ADSCs in a concentration- and time-dependent manner. The optimum inductive concentration of rhBMP-2 is 100 ng/mL, induced to 18-24 hours is a key period when rhBMP-2 is used to promote tissue engineering bone vascularization.
Objective To study the rule of lymphatic metastasis and to evaluate the extent of curative resection in advanced colorectal cancer. Methods One thousand and five lymph nodes from 114 consecutive patients with colorectal cancer underwent extended D3 resection were analyzed and classified as peritumor, longitudinal, and upward spread distribution. Results The metastatic rate and incidence of lymph node metastasis in peritumor, longitudinal as well as upward spread (N2 and N3) was 43.9% and 37.2%, 32.5% and 15.9% as well as 29.8% (19.3% and 10.5%) and 12.1% (16.6% and 7.8%) respectively. The distribution rate of metastatic lymph nodes was 17.5% and 23.5% in the longitudinal and upward spread respectively. In the longitudinal spread, most of lymph node metastasis was seen within 10 cm. Within 2 cm on the anal side in rectal cancer, the metastasis rate was 5.5%, and there was no metastasis in 2-4 cm. The lateral metastasis rate was 0%, 8.7% and 12.5% in the rectosigmoid (Rs), upper rectum (Ra) and lower rectum (Rb) respectively. Conclusion Advanced colorectal cancer tend to metastasize to longitudinal and upward lymph nodes. Jump metastasis is also a feature. In the lower rectal cancer within 6 cm from the anal verge or beyond pT3, there is a high risk of lateral metastasis. Extended D3 radical resection is necessary for colic cancer, but high ligation of the inferior mesenteric artery root as well as lateral lymphadenectomy and total mesenteric excision should also be performed for rectal cancer. There is no residual tumor tissue in the anastomosis when the excision distance is beyond 2 cm from the anal margin in rectal cancer.
This study was aimed to explore the features of clustered regularly interspaced short palindromic repeats (CRISPR) structures in Shigella by using bioinformatics. We used bioinformatics methods, including BLAST, alignment and RNA structure prediction, to analyze the CRISPR structures of Shigella genomes. The results showed that the CRISPRs existed in the four groups of Shigella, and the flanking sequences of upstream CRISPRs could be classified into the same group with those of the downstream. We also found some relatively conserved palindromic motifs in the leader sequences. Repeat sequences had the same group with corresponding flanking sequences, and could be classified into two different types by their RNA secondary structures, which contain "stem" and "ring". Some spacers were found to homologize with part sequences of plasmids or phages. The study indicated that there were correlations between repeat sequences and flanking sequences, and the repeats might act as a kind of recognition mechanism to mediate the interaction between foreign genetic elements and Cas proteins.
Finger tapping test is a common testing item for patients with Parkinson's disease (PD) in clinical neurology. It mainly evaluates the fine motor function of patient's hand in three aspects:amplitude, speed and regularity of the movement. This paper focused on the quantitative assessment of regularity of finger tapping movement for PD patients. The movement signals of thumb and index finger were recorded by using inertial sensor unit in the process of tapping test. Two nonlinear dynamic indexes, approximate entropy (ApEn) and sample entropy (SampEn), were calculated, and then the values were statistically analyzed. The experimental results indicated that both indexes had significant differences between patient group and control group. Moreover, the indexes had relatively high correlation with the scores of corresponding unified Parkinson's disease rating scale (UPDRS) item rated by clinical clinician, which illustrated that these two indexes could reflect the injury level of the repetitive finger movement. So, as a reliable method, it can be provided to the clinical evaluation of hand movement function for PD patients.
As pigs are similar to humans in anatomy, physiology and pathology, nutrition metabolism and disease characteristics, genetically modified pigs are already used for the studies of disease mechanism, pathology and toxicology and the evaluation of drugs. But the production of large modified animals is difficult, cumbersome, time-consuming and costly. With the breakthrough of gene editing technology, clustered regularly interspersed short palindromic repeat (CRISPR)/CRISPR-associated 9( Cas9)(CRISPR/Cas9) technology has greatly improved the mutation efficiency, reduced the cost and simplified the steps, and promoted the widespread application of genetically modified pigs. In this paper, the production methods of genetically modified pigs and the research progress of genetically modified pigs by CRISPR/Cas9 in the medical field were reviewed.
ObjectiveTo analysis the regularity of prescriptions of Chinese Medicine for syndrome of intense fire and heat (syndrome of Shi-Re-Huo-Du), and to provide evidences for the clinical treatment.MethodsCNKI, Sinomed, WanFang Data, VIP and Chinese medical databases were electronically searched to collect literatures about traditional Chinese Medicine for syndrome of Shi-Re-Huo-Du from inception to October, 2018. Two reviewers independently screened literature and extracted data. Then BICOMS 2 software was used to generate the co-occurrence matrix, NetDraw software was used to draw network maps.ResultsA total of 381 literatures involving 335 patients were included. There were 716 prescriptions of Shi-Re-Huo-Du (390 Chinese medicinal herbs), in which the frequently used herb included radix glycyrrhizae (346), Scutellariae Radix (255), Fructus Gardeniae (241), Rehmanniae Radix (239), Moutan Cortex (218), etc. The classification was performed according to traditional efficacy, Qing-re herbs (73), Bu-xu herbs (47), and Hua-tan-zhi-ke-ping-chuan herbs (39), accounted for 40.77% of the total herbs. The results of the analysis showed that the top herbs in Degree and Betweenness were consistent with high-frequency herbs. Among them, Rehmanniae Radix, Fructus Gardeniae, Scutellariae Radix, Moutan Cortex are the most important traditional Chinese herbs. High-frequency herbs are mainly based on Qing-re herbs, and the highest frequency of the herbs combination is Scutellariae Radix-Fructus Gardeniae.ConclusionAlthough there are kinds of herbs for syndrome of Shi-Re-Huo-Du, the commonly used herbs are relatively concentrated, and there is obvious interaction between high-frequency herbs, which is in line with the principle of compatibility of Chinese medicine. The high-frequency Chinese herbs and herb pairs were main components of the Chinese patent medicines and classic prescription. It reflected the rule of medicine used and essence in the treatment of syndrome of Shi-Re-Huo-Du.
ObjectiveTo explore and clarify the relationship between epileptic seizure and inducing factors. Avoid inducing factors and reduce epileptic seizure, so as to improve the quality of life in patients with epilepsy.MethodsClinical data of 604 patients diagnosed with epilepsy in Xijing Hospital of Air Force Military Medical University from January 2018 to January 2019 were collected. The clinical data of patients with epilepsy were followed up 6 months.ResultsAmong the 604 patients, 318 (52.6%) were seizure-free in the last 6 months, 286 (47.4%) had seizures. 169 (59.1%) had seizures with at least one inducing factor. Common inducing factors: 123 cases of sleep disorder (72.8%), 114 cases of emotion changes (67.5%), 87 cases of irregular medication (51.5%), 97 cases of diet related (57.4%), 33 cases of menstruation and pregnancy (19.5%), etc. Using the χ2 test, seizures with age, gender differences had no statistical significance (P > 0.05), but seizure type was statistically different between inducing factors. In generalized seizures, tonic-clonic seizures associated with sleep deprivation (χ2= 0.189), absence seizures and anger (χ2= 0.237), pressure (χ2= 0.203), irregular life (χ2= 0.214). In the focal seizures, focal motor seizures was correlated with coffee consumption (χ2=0.145), focal sensory seizures with cold (χ2=0.235), electronic equipment use (χ2 =0.153), satiety (χ2 =0.257). Complex partial seizures was correlated with anger (χ2 =0.229), stress (χ2 =0.187), and cold (χ2 =0.198). The secondarily generalized seizures was correlated with drug missing (χ2 =0.231), sleep deprivation (χ2 =0.158), stress (χ2 =0.161), cold (χ2 =0.263), satiety (χ2 =0.182). Among the inducing factors, sleep deprivation was correlated with anger (χ2 =0.167), fatigue (χ2 =0.283), and stress (χ2 =0.230).ConclusionsEpileptic seizure were usually induced by a variety of factors. Generalized seizures were associated with sleep disorders, emotional changes, stress, irregular life, etc. While focal seizures were associated with stress, emotional changes, sleep disorders, cold, satiety, etc. An analysis of the triggers found that sleep deprivation was associated with anger, fatigue, and stress. Therefore, to clarify the inducing factors of epileptic seizure, avoid the inducing factors as much as possible, reduce the harm caused by seizures, and improve the quality of life of patients.
The emergence of regular short repetitive palindromic sequence clusters (CRISPR) and CRISPR- associated proteins 9 (Cas9) gene editing technology has greatly promoted the wide application of genetically modified pigs. Efficient single guide RNA (sgRNA) is the key to the success of gene editing using CRISPR/Cas9 technology. For large animals with a long reproductive cycle, such as pigs, it is necessary to screen out efficient sgRNA in vitro to avoid wasting time and resource costs before animal experiments. In addition, how to efficiently obtain positive gene editing monoclonal cells is a difficult problem to be solved. In this study, a rapid sgRNA screening method targeting the pig genome was established and we rapidly obtained Fah gene edited cells, laying a foundation for the subsequent production of Fah knockout pigs as human hepatocyte bioreactor. At the same time, the method of obtaining monoclonal cells using pattern microarray culture technology was explored.
Objective To discuss the rules of using traditional Chinese medicine (TCM) to treat systemic lupus erythematosus (SLE) and Sjögren syndrome (SS). Methods The TCM prescriptions for patients with SLE or SS treated at the Department of Rheumatology of Longhua Hospital, Shanghai University of Traditional Chinese Medicine between January 2013 and July 2021 were collected. The prescriptions collected were broken down and analyzed for frequencies of the efficacies, properties, flavors and meridian tropisms of drugs in the prescriptions. The correlation and clustering analysis was performed in VOSviewer software. Results The medication information of 3689 cases of inpatients was included, from which 69167 pieces of data of using TCM prescriptions were extracted. The top 20 high-frequency drugs for treating SLE patients were mainly drugs for promoting blood circulation and removing blood stasis, qi-tonifying drugs, and drugs for heat clearing and detoxicating, which were mostly sweet, bitter or acrid in flavor, cold, warm or neutral in properties, and of the spleen, liver or stomach meridian. The drugs could be seperated into 3 clusters, the blue cluster mainly included qi-tonifying drugs, blood-tonifying drugs, and drugs for promoting blood circulation and removing blood stasis; the green cluster mainly included drugs for heat clearing and drugs for promoting blood circulation and removing blood stasis; and the red cluster mainly consisted of drugs for promoting blood circulation and removing blood stasis, drugs for inducing diuresis and alleviating edema, drugs for dispelling wind-heat, and digestant drugs. The top 20 high-frequency drugs for treating SS patients were mainly drugs for promoting blood circulation and removing blood stasis, qi-tonifying drugs, and drugs for inducing diuresis and alleviating edema, which were mostly sweet, bitter or acrid in flavor, slightly cold, cold or warm in properties, and of the spleen, stomach or liver meridian. The drugs could be seperated into 2 clusters, the green cluster mainly consisted of yin-tonifying drugs, drugs for promoting blood circulation and removing blood stasis, and drugs for heat clearing; the red cluster mainly included qi-tonifying drugs, drugs for promoting blood circulation and removing blood stasis, and drugs for inducing diuresis and alleviating edema. Conclusions The medication for SLE is in line with the treatment principle of “supplementing qi and nourishing yin, detoxicating and removing stasis”; the medication for SS, on the other hand, agrees with the basic rule of “nourishing yin and clearing away heat, promoting body fluid to moisten dryness”. Both are based on the approach of strengthening the body resistance, which is in line with the diagnostic and treatment ideas of rheumatology department and may offer the ideas of TCM syndrome differentiation for clinical use of drugs and thereby guiding the current clinical practice.