Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

In this work,a tetrahedral DNA nanostructure(TDN)functionalized rotational paper-based analytical device(RPAD)was constructed for rapid and highly sensitive detection of aflatoxin B1(AFB1)using exonucleaseⅠ(ExoⅠ)and G-quadruplex(G4)dimer.Herein,a single-stranded DNA,containing both of the G4 dimer sequence and AFB1 recognition sequence,was used as the recognition probe(G4 dimer probe).TDN was used to precisely regulate the orientation and distribution density of G4 dimer probe to improve the recognition efficiency of the system.ExoⅠas a single stranded DNA specific nuclease was introduced for effective amplification of the detection signal.G4 dimer was employed to enhance the fluorescence signal of thioflavin T(ThT).In the absence of AFB1,the G4 dimer structure of G4 dimer probe could specifically bind with ThT to generate dramatic fluorescence enhancement.However,in the presence of AFB1,AFB1 could specifically bind with G4 dimer probe,resulting in the dissociation of G4 dimer probe from TDN and further be digested by ExoⅠ.At the same time,the released AFB1 could bind to G4 dimer probe on the TDN again by this way to generate signal amplification.After this cycle,the amount of aptamer on the TDN was decreased,accompanied by the reduction of G4 dimer on TDN.In this case,the fluorescence intensity of the system was reduced.The designed RPAD showed a good linear response in AFB1 concentration range of 0.0001-500 ng/mL and the limit of detection was 0.1 pg/mL.Moreover,the proposed strategy was successfully applied to detection of AFB1 in peanut and wine.The developed TDN/G4 dimer/ExoⅠstrategy improved the specificity and sensitivity of the system significantly.

AIM: To explore the curative effect of modified Parks incision combined with rectus linear tucking on adults with concomitant strabismus and its influences on postoperative recovery of visual function and ocular aesthetics.METHODS: A total of 160 adults with concomitant strabismus were enrolled as the research subjects between June 2019 and June 2021. They were divided into observation group(modified Parks incision combined with rectus linear tucking, 89 cases)and control group(corneal limbal trapezoid conjunctival flap incision combined with rectus linear tucking, 71 cases)according to different surgical methods. The clinical curative effect and surgical related indexes between the two groups were compared. The changes in scores of corneal fluorescence staining, standard patient of eye dryness(SPEED), visual analogue scale(VAS)and Vancouver scar scale(VSS)before and after surgery in both groups were observed. The occurrence of complications such as poor conjunctival healing and astigmatism was statistically analyzed at 1mo after surgery.RESULTS: The total response rate of surgery in observation group was higher than that in control group, and length of surgical incision was shorter than that in control group(all P<0.05). The scores of postoperative corneal fluorescence staining and dry eye in observation group were lower than those in control group, and VAS scores at 1wk after surgery were lower than that in control group(all P<0.05). Postoperative divergent fusion range, strabismus angles of short-sighted 33cm and far-sighted 5m in observation group were greater than those in control group. Convergent fusion range was smaller than that in control group, and stereopsis of short-sighted 33cm and far-sighted 5m was lower than that in control group(all P<0.05). Postoperative break-up time in observation group was longer than that in control group, lacrimal secretion was more than that in control group, and scores of corneal surface regularity index(SRI)and VSS scores were lower than those in control group(all P<0.05). The total incidence of complications in observation group was lower than that in control group(P<0.05).CONCLUSION: The modified Parks incision combined with rectus linear tucking can not only improve surgical effect and promote the recovery of visual function and tear film function in adults with concomitant strabismus, but also relieve dry eye and postoperative pain, improve ocular aesthetics and reduce the incidence of postoperative complications.

ObjectiveTo investigate the dynamic change of monocyte-derived macrophages （MDMs） in chronic hepatitis B （CHB） during the treatment by ETV or TDF， and the association between the polarization of monocyte-derived macrophages （MDMs） and HBV DNA， HBeAg in CHB. MethodsPeripheral blood of 101 patients with chronic hepatitis B and 20 healthy donors were enrolled. MDMs were purified from 86 patients with CHB and 12 healthy donors by peripheral blood mononuclear cell （PBMC）. Flow cytometry was use to analyze the expression of specific markers on polarized macrophages to further evaluate the polarization of MDMs （M1/M2 MDMs）， including CD80， HLA-DR， CD163 and CD206. Additionally， qPCR was used to evaluate the mRNA expression of inflammatory factors of MDMs in 15 patients with chronic hepatitis B and 8 healthy controls. ResultsThe proportion of CD163⁺CD206⁺MDMs （M2 MDMs） were reduced in patients with CHB （P = 0.030）， compared with healthy controls. Correspondingly， the mRNA expression of anti-inflammatory factor， IL-10， decreased significantly （P = 0.040）. With the progress of nucleos（t）ide analogs （NAs） therapy， the proportion of CD163⁺CD206⁺MDMs （M2 MDMs） increased gradually （P < 0.001）， while the proportion of CD80⁺HLA-DR⁺MDMs （M1 MDMs） decreased gradually （P = 0.005）. In addition， M2 MDMs were negatively associated with both HBeAg （P = 0.019）， HBV DNA （P = 0.002）， AST （P = 0.048） and ALT （P = 0.030） in patients with CHB. ROC curve analysis showed that CD163⁺CD206⁺MDMs （M2 MDMs） had high predicting value for the virologic response of CHB during NA therapy， and could be used to predict undetectable HBV DNA ［ROC 95％CI： 0.705 （0.594，0.815）］ and HBeAg seroconversion ［ROC 95％CI： 0.740 （0.634，0.847）］. ConclusionsThere are dynamic changes of MDMs polarization in patients with CHB during NA therapy， and the negative correlation between the proportion of M2 MDMs and HBV DNA， HBeAg indicates that M2 MDMs could have a high predicting value in the virologic response of CHB during NA therapy.

Aim To evaluate the effects of androgra- pholide ( Andro) on suppressing tumor growth and improving mitochondrial function on mouse breast cancer and explore its mechanism.Methods MTT assay was performed to measure the effect of Andro on the growth capacity of mouse breast cancer cell line 4T1.Mice were treated with Andro, then tumor volume measured, organ index calculated and Hematoxylin - eosin ( HE ) stained to detect the inhibitory effect of Andro on tumors.Lactate assay kit was used to detect the lactate level in mouse serum to check glycolysis discrepancy.To illustrate the transformation in oxidative phosphorylation ( OXPHOS) , Acetyl coenzyme A ( Acetyl-CoA ) assay kit was used to ascertain Acetyl-CoA content level , Western blot was used to detect the protein expression of pyruvate dehydrogenase ( PDH) , and ATP assay kit was used to ascertain ATP content level.The mitochondria functions were analyzed: oxygen ( ()2 ) consumption was measured by Clark oxygen electrode, and mitochondrial membrane potential was detected by JC-1 staining.Results Andro could effectively inhibit the proliferation of 4T1 cells and the growth of tumors, and had no significant damage on normal organs.An- clro reduced serum lactic acid content, indicateing that Andro inhibited the process of glycolysis.The expression of PDH, content of acetyl-CoA and ATP content increased with the increase of Andro concentration.It showed that Andro could up-regulate oxidative phosphorylation.In addition, the ()2 consumption and mitochondrial membrane potential increased in 4T1 cells, indicating that Andro could recover mitochondrial function.Conclusions Andro can inhibit the growth of mouse breast cancer, and its mechanism may be related to the inhibition of glycolysis level, restoration of OXPHOS and improvement of mitochondrial function.

As the most advanced environment-friendly production model in the international society, ecological agriculture of Chinese materia medica(CMM) is the only way for the development of modern agriculture. With the proposal of the declaration on ecolo-gical agriculture of CMM, "Don't grab land from farmland, don't be enemies of grass and insects, don't be afraid of barren slopes and forests, and live up to the green and green mountains", the ecological planting of CMM has blossomed all over the country, and formed a scientific theory, technology and model. Based on the theory and method of economics, this paper expounds the comprehensive benefits and development advantages of ecological agriculture of CMM from the perspectives of farmers(producers), patients(consumers) and the country. From the perspective of medicinal farmers, the input and output income of conventional agriculture and ecological agriculture of CMM such as Panax ginseng, Astragalus propinquus, Atractylodes lancea, and Bupleurum chinense were compared, and it was found that ecological agriculture of CMM had obvious advantages in net income, average annual income and input-output ratio, which could better promote farmers' income. From the perspective of patients, according to the same dose, the content of active ingredients in ecologically planted CMMs is significantly higher than that in conventionally-planted herbs, and the amount of effective substances taken by patients is also higher, so as to achieve better therapeutic effect. At the national level, ecological planting of CMM is the key to ensuring the high-quality development of CMM industry, increasing farmers' income, ensuring the safety of people's drug use and promoting the sustainable development of agriculture. It is also an important part of realizing the harmonious development of economy, society and environment and promoting ecological civilization. In general, the declaration on ecological agriculture of CMM embo-dies the core characteristics and goals of ecological agriculture, and also points of the path and vision of ecological agriculture of CMM in the future. The declaration will guide production practice, promote the benefit of farmers, and lay the foundation for the sustainable development of CMM industry.
Agriculture ; Drugs, Chinese Herbal ; Humans ; Materia Medica ; Medicine, Chinese Traditional ; Plants, Medicinal

Aortic Valve/surgery* ; Aortic Valve Stenosis/surgery* ; China/epidemiology* ; Heart Valve Prosthesis Implantation ; Humans ; Severity of Illness Index ; Treatment Outcome

OBJECTIVE: To investigate the clinical characteristics, laboratorial and bone marrow pathological features of primary thrombocytopenia (ET) patients with different mutations of CALR, JAK2 and MPL genes. METHODS: The chinical data of 120 cases of ET in Jiangsu provincial people's hospital/ The First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2017 were collected and analyzed, including 76 cases with JAK2 gene mutation, 40 cases with CALR gene mutation, 2 cases with MPL gene mutations, 2 cases without gene mutation. RESULTS: Among the ET patients, compared with the JAK2 gene mutation, CALR gene mutation showed statistically significant deareament of white blood cells and hemoglobin (P=0.001, P=0.01) and the male platelets in CALR group showed significant increament (P=0.04). Fourthermore, the average number of megakaryocytes and its cluster numbers in each hight power field of vision showed statistically significant decreament in CALR group as compared with JAK2 group (P=0.001, P=0.001), and thrombotic events in CALR group were signicantly lower than those in JAK2 group (7.5% vs 18.4%) (P=0.03). CONCLUSION Mutations of CALR, JAK2 have different clinical characteristics and blood pathological changes of Chinese ET patients, and their clinical significance is worth to explore.
Bone Marrow ; Calreticulin ; genetics ; China ; Humans ; Janus Kinase 2 ; genetics ; Male ; Mutation ; Receptors, Thrombopoietin ; genetics ; Thrombocythemia, Essential

The cluster brand is the embodiment of the core competitiveness of an industry. Developing and cultivating cluster brand of ecological agriculture of Dao-di herbs not only helps to optimize the value chain of the Chinese medicinal materials(CMMs) industry cluster, realize the value-added of the CMMs industry cluster, but also enhance the visibility and influence of the industrial cluster, enhance the core competitiveness of the industrial cluster. This has important practical significance for promoting the "orderly" "safe" and "effective" development of the Dao-di herbs. Based on the industry development status of CMMs, this article introduces several concepts related to cluster brands and their relationships, and focuses on the cultivation models and strategies of cluster brand in the CMMs industry. Based on the current status of the development of the CMMs industry, this article introduces several concepts related to cluster brands and their interrelationships. It discusses the cultivation models and strategies of cluster brands in the CMMs industry, industry associations, Chinese medicine companies and individual growers as the support, insists on the ecological cultivation of authentic medicinal materials and the cultivation of cluster brands. Finally, it points out the direction for the high-quality development of the ecological agriculture of CMMs.
Agriculture ; Drugs, Chinese Herbal ; Industrial Development ; Medicine, Chinese Traditional ; Plants, Medicinal

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires