Objective:To investigate mechanisms of IL-17D on cytotoxic function of CD93+CTL in lung tumor microenviron-ment(TME)and promotion of Platycodon grandiflorum(PG).Methods:Lung of B16 melanoma model mice and control mice were treated with PG or IL-17D,and lung tumor clone formation was observed.IL-17D expression change in lung was detected by single cell sequencing,immunofluorescence staining and flow cytometry.Single cell sequencing and flow cytometry were used to detect CD93+CTL content,cytotoxic phenotype and functional factors changes,including CD107a,perforin,granzyme B,chemokine CCL2,CXCL9 and their ligand CCR2 and CXCR3.Western blot and RT-PCR were used to detect effect of Platycodonopsis saponin D on IL-17D and its transcriptional regulator NRF2 expressions in EL4 cells.Results:Pulmonary CD93+CTL highly expressed cytotoxic effectors such as perforin and CD107a and chemokine receptors CXCR3 and CCR2 than CD93-CTL,while there was no significant difference in secretion of granzyme B.In mouse lung tumor model,pulmonary IL-17D and CD93+CTL were significantly decreased(P<0.001);in tumor-bearing mice after IL-17D backfill assay,or after 10 days of treatment with PG,proportion and absolute number of IL-17D and CD93+CTL in lungs were significantly increased(P<0.05),and tumor clones were significantly reduced;meanwhile,tumor-local expressions of cytokines CCL2,CXCL9,which are related to recruitment and function of CD93+CTL,and IL-17D were significantly upregulated.Up-regulation of IL-17D and its transcriptional regulator NRF2 by PG was verified in vitro experiments on EL4 cell line by PD.Conclusion:Traditional Chinese medicine PG and its extracts can up-regulate expression of IL-17D in lungs,improve infiltration and cytotoxic function of CD93+CTL and antagonize malignant progression of lung tumors,this is an important phar-macological mechanism of PG in improving immune TME of tumors in lung.

OBJECTIVE: To explore the molecular etiology for a Chinese family with mitochondrial DNA depletion syndrome. METHODS: Genomic DNA was extracted from peripheral blood samples of the patient and her parents.Targeted capture and next-generation sequencing was carried out to detect potential variants. Suspected variant was validated by Sanger sequencing. RESULTS: A novel homozygous frameshift variant c.505_508delTATC was identified in the patient, for which both his mother and father were carriers. CONCLUSION The frameshift variant c.505_508delTATC probably underlies the mitochondrial DNA depletion syndrome in this patient. The result also enriched the variant spectrum of DGUOK gene.
Asian Continental Ancestry Group ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Frameshift Mutation ; Humans ; Mutation ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; Syndrome

Objective? To explore the application effect of self-help lower limb functional exercise shoes in elderly patients after total knee arthroplasty(TKA). Methods? A total of 58 patients with knee osteoarthritis who underwent TKA in the Second Affiliated Hospital of Wenzhou Medical University from September of 2017 to September 2018 were selected and randomly divided into the control group(n=28) and observation group(n=30). Routine functional exercise method was adopted in the control group while self-help lower limb functional exercise shoes were applied in the observation group. The time needed to master the functional exercise instruction, compliance with the functional exercise and knee joints activity post-operation in the two groups were compared. Results? The time needed to master the functional exercise instruction in the observation group was (4.3±1.5)min, shorter than the control group with statistical significance (t=4.87, P＜ 0.01). In the observation group there were 27 patients who had high compliance with the functional exercise, while 15 patients in the control group had high compliance, the difference was statistically significant (χ2=9.62,P＜0.05). Three months after operation, the knee joint activity of the observation group was better than that of the control group (115.3±11.3)°, the difference was statistically significant (t=2.45, P＜0.05). Conclusions? The use of self-help lower limb functional exercise can help the elderly TKA patients to master the functional exercise method faster, improve their compliance with postoperative functional exercise, improve postoperative rehabilitation exercise effects and promote early recovery of the affected limb functional.

Objective To explore the effects and mechanism of electroacupuncture (EA) on denervation-induced atrophy in rats. Methods A total of 18 male Sprague-Dawley rats were divided into sham group (n=6), model group (n=6) and EA group (n=6). The latter two groups were clamped right sciatic nerve to establish atrophy model of skeletal muscle. On the second day after modeling, EA group accepted electroacupuncture on right Zusanli (ST36) and Huantiao (GB30) for two weeks. Their gastrocnemius muscles were obtained after intervention, and the wet weight ratio of the gastrocnemius muscles was calculated. The cross-sectional area (CSA) and diameter of muscle fibers were measured after HE staining. The protein expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), 70-KD ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6k (p-p70S6k) was tested with Western blotting. The gene expression of mTOR and p70S6K was detected with real-time quantitative polymerase chain reaction. Results Compared with the sham group, the wet weight ratio of the gastrocnemius muscle, CSA and diameter of the muscle fibers decreased in the model group and EA group (P<0.001), which were more in EA group than in the model group (P<0.01); the protein expression of mTOR, p-mTOR, p70S6K and p-p70S6K increased in the model group (P<0.01), and increased more in EA group (P<0.05); the gene expression of mTOR and p70S6K increased in the model group (P<0.05) , and increased more in EA group (P<0.05).Conclusion Electroacupuncture delays the atrophy of denervated skeletal muscles, which may relate to activation of mTOR/p70S6K signal pathway to impact synthesis of skeletal muscle proteins.

Objective To explore the risk factors of postoperative activities of daily living in elderly patients after hip fracture. Methods By convenience sampling,from October 2013 to June 2015,150 elderly patients having undergone surgery after hip fracture were selected,with data of 141 cases finally collected. Gender,age,underlying diseases,surgical procedures,postoperative complications,ambulation time,whether walking with aids,family caregivers and reexamination conditions were recorded for evaluation of the patients about their activities of daily living half a year after the surgery. Mean comparison and chi-square test were used for univariate analysis about the above factors and Logistic regression analysis was used to identify possible risk factors. Results It was shown in univariate analysis that recovery of postoperative activities of daily living was influenced by age,surgical procedures,postoperative complications,ambulation time,walking aids,etc. (P< 0.05);it was shown in multivariate logistic regression that age (OR=1.065,95%CI:1.010-1.124, P=0.020),surgical procedures (OR=2.900,95%CI:1.200-7.008,P=0.018),postoperative complications (OR=6.366,95%CI:2.592-15.632,P< 0.001),ambulation time (OR=2.684,95%CI:1.133-6.359,P=0.025) and walking aids (OR=5.796,95%CI:1.542-21.780,P=0.009) were risk factors of postoperative daily living ability in elderly patients after hip fracture. Conclusions Old age,developing internal fixation surgery after incision,postoperative complications, walking more than 3 months after operation or walking without aids affect postoperative activities of daily living in elderly patients after hip fracture,thus affecting their quality of life, which makes it necessary for nursing staff to focus on risk factors for positive prevention and intervention.

AIM:To observe the effect of plumbagin on the mRNA and protein expression of nicotinamide ade -nine dinucleotidephosphate oxidase 4 ( Nox4 ) , reactive oxygen species ( ROS ) level and protein expression of α-smooth muscle actin (α-SMA) in the HSC-LX2 cells stimulated with transforming growth factor β1 (TGF-β1) in vitro.METH-ODS:HSC-LX2 cells were cultured in vitro and divided into blank group, model group, high-, medium-and low-dose (2, 1.5 and 1 μmol/L) plumbagin groups .After incubated with each drug for 72 h, the mRNA expression of Nox4 was detec-ted by RT-PCR.ROS levels were tested by in situ loading probe method.The protein contents of Nox4 and α-SMA were measured by Western blot .RESULTS:Compared with model group , after treated with plumbagin for 72 h, the mRNA ex-pression of Nox4, ROS level and α-SMA protein were significantly decreased in high-and medium-dose plumbagin groups (P<0.01).CONCLUSION:Plumbagin inhibits the activation of HSC-LX2 cells via decreasing the expression of Nox4, thus decreasing ROS levels .

Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N '-( 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl)-bis(2-( pyrroli-din-1-yl)acetamide) (XP-16) on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2+ concentration ( [ Ca2+] i ) and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A ( MT-1 A ) transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction ( qRT-PCR) . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic mor-
phology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2+] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2+ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2+] i .

AIM: To study the exciting effects of ethanol extract of capitate phrynium (Phrynium Capitatum Willd) stem and leaf on the uterine smooth muscle of rat,mice,guinea pig and rabbit pretreated by diethylstilbestrol. METHODS: Conventional method was adopted to prepare animal uterine specimens both in vivo and in vitro. Different doses of the extract were given to the specimens and recorded their tensility, frequency and activity of contraction with the help of BL-420 biological experimental system. IC_(50) of the extract was calculated, with oxytocin as standard comparison. RESULTS: 95% ethanol extract and 50% ethanol extract of capitate phrynium and leaf had obviously stimulating action on the uterine smooth muscle of rat, mice, guinea pig and rabbit in a dose-dependent manner in vitro, whichinclude accelerating contraction frequency, raising tension (P<0.05, P<0.01). In addition, 50% ethanol extract also had a significant stimulating action on uterine smooth muscle of rabbit in vivo(P< 0.01). CONCLUSION: Ethanol extracts of capitate phrynium have an effect on rodent uterine. Smooth muscle indicates that clinical application of Chinese practitioner used to treat the dysfunctional uterine bleeding proves to be effective.

Objective To investigate the apoptotic and proliferation effects of signal transduction inhibitors on human endometrial carcinoma cells with different PTEN gene expression. Methods FTEN antisense oligonucleotide and pcDNA3.1/PTEN vector contained PTEN gene were transfected into endometrial carcinoma cells (HEC-1A and Ishikawa). The expression of PTEN protein was detected by confocal spectral microscopy. The endometrial carcinoma cells (HEC-1A, HEC-1A-PTEN-null, Ishikawa, Ishikawa-PTEN) were treated with signal transduction inhibitors, RG-14620, SB203580 (SB) and rapamycin, respectively. Cell apoptosis morphology, cell apoptosis and cell cycle were detected by Hoechst 33258 staining and flow cytometry. Cell viability was determined by methyl thiazolyl tetrazolium assay. Results The PTEN protein expression in two endometrial carcinoma cells (Ishikawa, HEC-1A) was exchanged by PTEN antisense oligonucleotide blocked and pcDNA3. 1/PTEN stable transfected. After treated with RG-14620, SB and rapamycin, marked morphological changes of apoptosis were observed in HEC-1A-PTEN-null and Ishikawa cells. The cell apoptosis of HEC-1A-PTEN-null and Ishikawa cells exposed to SB were significantly increase [(31.6±0.8)% and (37.8±0.8)%, respectively], the G1 phase cells were increased to (84.1±3.2)% and (87.5±1.9)%. While cell viability was significantly decreased in HEC-1A-PTEN-null and Ishikawa cells, the cell viability of HEC-1A-PTEN-null and Ishikawa cells exposed to SB were (54.0±2.1) % and (49.0±1.7) %. Conclusion Loss of PTEN in endometrial carcinoma cells may improve the G_1 phase cells and apoptotic effects, inhibit the cell proliferation, which due to the sensitivity of cells to related signal transduction inhibitors.

BACKGROUND: The essential component of Plumbago zeylanica Linn is plumbagin, which plays a role in anti-leukemia, antibiosis, antifertility, as well as cardiovascular. OBJECTIVE: To investigate the effects of Plumbago zeylanica Linn on the healing process of experimental rabbit fracture. DESIGN, TIME AND SETTING: Randomized control experiment of animal was performed at the Department of Pharmacology, Guilin Medical College between December 2006 and February 2007. MATERIALS: Eight New Zealand rabbits, 80 to 85-day-old, weighing 2.1-2.5 kg, half male and half female. METHODS: Eight rabbits were randomly divided into the control and experimental groups, with 4 animals in each group. Under sterile conditions, the experimental bone fracture model of rabbit was created by making a longitudinal incision in the medial of the middle portion of right foreleg, cutting off a point-foot radius with bone clamp, and suturing the incision. The rabbits in the experimental group were treated externally with the extraction of Plumbago zeylanica Linn in 50% of ethanol, twice a day for 15 days. The sodium chloride was administrated into rabbits in the control group. MAIN OUTCOME MEASURES: The serum alkaline phosphatase (S-ALP), Ca2+, P3+, K+, Na+, Cl-, liberation and total calcium of the rabbit were measured at 15 and 30 days after drug administration, X-ray of the bones were performed at the fracture healing. RESULTS: Compared with the control group, the fracture healing of the experimental group was faster, and the serum examinations showed that the serum Ca2+ levels of the experimental group was significantly increased by Plumbago zeylanica Linn, there had significantly differences between prior to and after drug administration (P