Object To study the effects of different growth regulators and organic additives on rooting of regenerated shoots from Dendrobium huoshanense C Z Tang et S J Cheng protocorm like bodies Methods In order to induce root development, the regenerated shoots were cultured on MS medium containing 3% sucrose and 0 8% agar and supplemented with NAA, IBA, BA, KT, banana mud or amino acids at different concentrations During rooting induction, the development of roots was observed and the frequency of rooted shoots was scored Results The frequency and the process of root initiation were significantly influenced by different growth regulators or organic additives For initiation and development of roots, the suitable concentration of NAA, IBA, BA, KT, banana mud or amino acids was 0 2 , 0 1, 0 8, 0 8 mg/L, 20% and 0 1%, respectively Conclusion In all media tested, NAA, IBA or banana mud display greater effects on rooting of regenerated shoots of D huoshanense than others and KT enables shoots to keep green

Objective:To explore the relationship between angiotensin converting enzyme (ACE) gene insertion/dele‐tion (I/D) polymorphism and familial essential hypertension (EH) .Methods :Fluorescent in situ hybridization stai‐ning DNA sequencing test was used to detect ACE gene I/D genotype frequency and allele frequency in 46 familial EH patients (familial EH group) ,64 EH patients without family history (non‐familial EH group) and 43 healthy people (healthy control group) .Results:In healthy control group ,non -familial EH group ,familial EH group ACE gene DD genotype frequency was 11.6% ,32.3% and 37.0% ;distribution frequency of D allele was 33.7% , 52.3% and 57.6% respectively .Compared with healthy control group ,there were significant rise in ACE gene DD genotype and D allele frequency in familial EH group and non -familial EH group (P<0.05 or <0.01) ,but there were no significant difference between familial EH group and non‐familial EH group (P>0.05) .Conclusion:ACE gene DD genotype and D allele may be genetic predisposing genes of patients with essential hypertension ,and there is no significant difference in genetic constituent ratio between familial EH patients and non‐familial EH patients .

Objective To study the effect of overexpression of TRPC6 on Ang Ⅱ-induced apoptosis of mouse podocytes in vitro and to explore the possible mechanisms. Methods Mouse TRPC6 cDNA eukaryotie expression vector pEGFP-NI-mTRPC6 was transfected to conditionally immortalized routine podocyte cell line by liposome. The fluorescent microscopy was used to examine the expression of EGFP after 24 hours. The change of TRPC6 protein expression was observed by Western-blot. Podocytes were treated by different concentrations of Ang Ⅱ. The podocyte intracellular calcium concentration was measured with laser-scanning con_focal microscope. The expression of Bax and Bcl-2 mRNA was assessed by RT-PCR and the expression of Bax and Bcl-2 protein was measured by Western-blot. The apoptotic ratio of podocytes was monitored by flow cytometry and Hoechst staining. Results About 35% of the cells expressed EGFP. An up-regulation of protein expression of TRPC6 was detected in podocytes when transfected with pEGFP-N1-mTRPC6 (P<0.01). The overexpression of TRPC6 promoted the Ang Ⅱ-induced influx of extracellular calcium and elevated the expression of Bax but decreased the expression of Bcl-2 (P<0.01, P<0.05). The apoptotic ratio of podocyte was (2.50±0.72)% when treated by low-dose Ang Ⅱ (10-10 mol/L), and it was increased to (4.33±0.45)% when transfected with pEGFP-N1-mTRPC6 (P <0.05 ). Transfection with pEGFP-NI-mTRPC6 increased apoptosis rate from (15.46± 1.40)% to (18.33±0.87)%(P<0.01) by high-dose Ang Ⅱ (10-6 mol/L). Conclusion TRPC6 plays an important role in the Ang Ⅱ-induced apoptosis of podocytes by promoting the influx of extraeellular calcium, which leads to the apoptosis cascade initiation.

AIM: To explore the inhibitory effect of oxymatrine (OMT) on the allergic contact dermatitis (ACD) stimulated by dinitrofluorobenzene (DNFB) and its effects on the proliferation of the lymphocytes. METHODS: ① An ACD mouse model was established by stimulation with DNFB, and then the mice were injected intraperitoneally with different dosages of OMT, PBS and hydrocortisone (HCT) respectively, the swelling degree of their auricles was examined. ② Carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE) dye and flow cytometer were used to examine the fluorescence intensity changes of lymphocytes stimulated by polyclonal stimulator ConA and OMT. RESULTS: ① compared with PBS group, OMT possessed the strong inhibitory effect on the ACD caused by DNFB in a dose-dependent manner, and its inhibitory effect was equivalent to the HCT of the same dosage with fewer side effects. ② In vitro experiments proved that OMT (500, 125 and 31 mg/L) had the ability to restrain the proliferation of lymphocytes of mouse. CONCLUSION: OMT possesses an inhibitory effect on the ACD induced by DNFB, and OMT is a kind of immunosuppressor.

ObjectiveTo induce human peripheral blood mononuclear cells differentiate into hepatocyte-like cells by hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro and determine whether PKH26 could be used to serve as an effective tracer for the cells,and observe the ability of transplanted hepatocyte-like cells differentiate into hepatic cells in nude mice.MethodsGroup A and B were set up respectively.In Group A,mononuclear cells were cultivated without hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in cell culture.They were used as negative control group.In Group B,mononuclear cells were cultured with the administration of both HGF and FGF-4 to induce the differentiation into liver hepatocyte-like cells.The changes in cell morphology were observed and the expressions of AFP and CK 19 were detected by immunocytochemical staining in two groups at different times after induction.The hepatocyte-like cells differentiated from human peripheral blood mononuclear cells labeled by the fluorescent dye PKH26 injected into caudal vein in nude mice is experimental group.The nude mice injected with equal amount of normal saline in control group.The migration of the labeled cells into the liver are observed by the fluorescence microscope in the hepatic tissue sections of nude mice and the expressions of ALB were detected by immunocytochemical staining two weeks after the cells transplantation.ResultsCells in group B have a strong proliferative activity.It becomes large and oval,grows in colonies following induction.Cells in group A that showed spherical shape when peripheral blood mononuclear cells were just isolated are gradually becoming inconformity in morphology,spindle or fibroid,and a few cells are round:cells developed apoptosis and cracked following incubation.The expressions of AFP and CK19 were positive after induction in group B as detected by immunocytochemicat staining.Inversely,the expressions of AFP and CK19 were negative in group A after incubation.The experimental group showed numerous PKH26 labeled cells in the hepatic tissue sections of nude mice.But the control group did not show PKH26 labeled cells.The expressions of ALB were positive in the experimental group as detected by immunocytochemical staining after two weeks of the cells transplantation.ConclusionHuman peripheral blood mononuclear cells have the potential to differentiate into hepatocyte-like cells under the induction of HGF and FGF-4.Additionally,PKH26 is an effective tracer in hepatocyte-like cell transplantation.The hepatocyte-like cells settled in hepatic tissue begin to differentiate into mature hepatocyte after two weeks of the cells transplantation.It plays hepatic cells function and expresses alhnmin.

Making bone scaffold through tissue engineering method presents a new choice for both the patients and the doctors of orthopaedics. The biodegradable polymer PLA is chosen to make porous fundus scaffold jetting through special designed nozzle on multi-functional rapid prototyping machine controlled by computer according to the CT data CAD model. The scaffold is then chemically aggregated to compound with collagen-hydroxyapatite, and the ideal bone repair material is obtained. Animal experiment has indicated the correctness of this conclusion.
Absorbable Implants ; Animals ; Biocompatible Materials ; chemistry ; Bone and Bones ; Collagen ; chemistry ; Durapatite ; chemistry ; Humans ; Lactic Acid ; chemical synthesis ; Polyesters ; Polymers ; chemical synthesis ; Porosity ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective:To discuss the effect of pharmacist intervention on the use of antimicrobial agents in the clinical pathway of community-acquired pneumonia ( CAP) in our hospital to standardize the rational medication and promote the rational use of antimicro-bial agents. Methods:Totally 100 bacterial CAP patients in 2013 ( before the intervention) and 2014 ( after the intervention) in the pneumology department were studied. The antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospi-talization time, therapeutic effects and so on in the two groups were observed during the treatment. Results: There were significant differences between the two groups in antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospitaliza-tion time and so on, whereas there was no significant difference in the curative effect. Conclusion:After the pharmacist intervention, the application of antibiotics is more rational, the antibiotics use density and per capita cost are reduced, the hospitalization day is shortened and the value of pharmacists is also improved.

Cartilage has poor self-recovery because of its characteristics of no blood vessels and high extracellular matrix. In clinical treatment, physical therapy or drug therapy is usually used for mild cartilage defects, and surgical treatment is needed for severe ones. In recent years, cartilage tissue engineering technology provides a new way for the treatment of cartilage defects. Compared with the traditional surgical treatment, cartilage tissue engineering technology has the advantages of small wound and good recovery. The application of microcarrier technology in the design of tissue engineering scaffolds further expands the function of scaffolds and promotes cartilage regeneration. This review summarized the main preparation methods and development of microcarrier technology in recent years. Subsequently, the properties and specific application scenarios of microcarriers with different materials and functions were introduced according to the materials and functions of microcarriers used in cartilage repair. Based on our research on osteochondral integrated layered scaffolds, we proposed an idea of optimizing the performance of layered scaffolds through microcarriers, which is expected to prepare bionic scaffolds that are more suitable for the structural characteristics of natural cartilage.
Cartilage ; Extracellular Matrix/chemistry* ; Technology ; Tissue Engineering/methods* ; Tissue Scaffolds/chemistry*

OBJECTIVE: To assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies. METHODS: 10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13. RESULTS: The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly. CONCLUSION The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.
Pregnancy ; Female ; Humans ; Trisomy/genetics* ; Retrospective Studies ; Prenatal Diagnosis/methods* ; Down Syndrome/genetics* ; Aneuploidy ; Chromosome Aberrations ; Trisomy 18 Syndrome/genetics* ; Sex Chromosome Aberrations ; Fetus

OBJECTIVE: To carry out prenatal diagnosis for a fetus with mosaicism Yq deletion. METHODS: A fetus with high risk of sex chromosomes indicated by non-invasive prenatal testing (NIPT) at Tianjin Medical University General Hospital in July 2021 was selected as the study subject. Prenatal diagnosis of the fetus was performed with combined G-banded chromosomal karyotyping, fluorescence in situ hybridization (FISH), copy number variation sequencing (CNV-seq), real-time fluorescence PCR (QF-PCR), and ultrasound examination. RESULTS: Analysis of the amniocytes at 23 gestational weeks had yielded a 45,X karyotype. However, FISH had shown signals of Y chromosome. Re-examination by cordocentesis had shown a mosaicism of 46,X,+mar[33]/45,X[17]. FISH showed that 69% of the cells had contained Y chromosome signals. The result of CNV-seq was seq[19]del(Y)(q11.1q12)(mos) chrY: g.13200001_ 28820000del (mosaicism rate = 64%), which suggested mosaicism for a Yq deletion, which encompassed the azoospermia factor (AZF) region. Deletion of the AZF region was verified by QF-PCR. The fetal karyotype was ultimately determined as mos46,X,del(Y)(q11.1)[33]/45,X[17]. Although ultrasound examination had shown no abnormality in the fetus, the couple had opted to terminate the pregnancy, and the induced fetus had a normal male appearance. CONCLUSION The combined use of multiple techniques is beneficial for accurate and rapid prenatal diagnosis. For fetuses with mosaicism chromosomal abnormalities, it may be difficult to accurately predict the postnatal phenotype. It is therefore necessary to further explore their genotype-phenotype correlation in order to provide better guidance upon genetic counseling.
Female ; Pregnancy ; Male ; Humans ; Mosaicism ; DNA Copy Number Variations ; In Situ Hybridization, Fluorescence ; Y Chromosome ; Fetus