BACKGROUND: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) gene showed low expression in glioma cells. LRIG1 gene overexpression significantly enhanced LRIG1 mRNA and protein expression and inhibited its biological behavior. However, very few researchs are reported from the stand-point of inhibition of LRIG1 gene expression. OBJECTIVE: To construct specific RNA interference plasmids for LRIG1, establish stably transfected human glioma GL15 cell line, and observe its effect on expression of target gene LRIG1. METHODS: Designed and synthesized two shRNAs (named LRIG1-shRNA1 and LRIG1-shRNA2) specific for LRIG1 mRNA according to the GenBank, and one scrambled shRNA sequence as negative control, named pGenesil2-negative shRNA. The shRNA was inserted into pGenesil2 vector and sequenced. The recombinant vectors were transformed into E. coli. Picked up the positive clones and extracted the plasmids, which were transfected into GL15 cells by Metafectine. G418 was applied to select the stably transfected cell clones. Western Blotting was performed to examine the LRIG1 protein level.RESULTS AND CONCLUSION: The recombinant plasmids which contain shRNA were analyzed by restriction endonuclease digestion and DNA sequence, and it was proved that the fragment was inserted into the expected sites. Compared with the negative control group, the level of LRIG1 protein expression in pGenesil2-LRIG1-shRNA1(LRIG11) transfected cells and in pGenesil2-LRIG1-shRNA2(LRIG12) transfected cells was decreased by 47.9% (P < 0.01) and 32.8% (P > 0.05). The results confirmed that RNAi expressing vector specific for LRIG1 gene (pGenesil2-LRIG1-shRNA1) was successfully constructed, and the stable cell clones transfected with the shRNA expression vector showed inhibition of the expression of LRIG1 in glioma cell line GL15.

Objective To investigate the antitumor effects and the possible mechanisms of dendrit-ic cells co-cultured with cytokine-induced killer cells(DC-CIK)in combination with sorafenib on two lung adenocarcinoma cell lines,A549 cells(harboring KRAS gene mutation)and PC-9 cells(harboring EGFR gene mutation). Methods DC and CIK cells were routinely generated in vitro by stimulating PBMCs isola-ted from lung cancer patients with different cytokines and then co-cultured after a week of culturing. Flow cy-tometry analysis(FCM)was used to analyze the phenotype of DC-CIK cells after 7 days of co-culturing. The 50% inhibitory concentration(IC50 )of sorafenib against tumor cells was detected by MTT assay. The tumor cells were treated with DC-CIK cells alone or in combination with sorafenib. The proliferation of tumor cells was tested by CCK-8 kit and dynamically monitored by real-time cellular analysis(RTCA). Annexin-V/ PI staining was used to examine the apoptosis rates in each group. Real-time fluorescent quantitative PCR and FCM were respectively performed to detect the expression of natural killer group 2 member D ligands (NKG2DLs)at mRNA and protein levels after the treatment with sorafenib for 24 h. Results There was no significant difference between the IC50 of sorafenib against A549 and PC-9 cells after a 24-hour exposure(P﹥0. 05). Compared with the DC-CIK biotherapy,treating the tumor cells with DC-CIK cells in combination with sorafenib significantly inhibited the cell proliferation and increased the total apoptosis rates of tumor cells(P﹤0. 05). Moreover,the inhibition rates to tumor cell proliferation were enhanced along with the in-crease of effect-to-target ratio(E/ T). Compared with the single-factor treatment groups,the normalized cell index(NCI)in the combined treatment group was significantly decreased. Blocking NKG2D could abate the inhibitory effect of DC-CIK cells on tumor cell proliferation(P﹤0. 05). The expression of NKG2DLs(inclu-ding ULBP1,UBLP2 and ULBP3)on tumor cells at mRNA and protein levels were increased to different ex-tent after treating with 5 μmol/ L of sorafenib for 24 h. Conclusion There was no significant different be-tween the inhibitory effects of sorafenib on the proliferation of lung adenocarcinoma cancer cells harboring KRAS or EGFR gene mutation. The antitumor effects of DC-CIK cells combined with sorafenib on lung ade-nocarcinoma cells might be induced by regulating the NKG2D-NKG2DLs pathway and enhancing apoptosis. Moreover,the antitumor effects of the combined treatment were better than those of single-factor treatments.

Objective To evaluate the functional changes of visual cortex (V1) in patients with primary open angle glaucoma (POAG) by fMRI retinotopic mapping technology. Methods Fifteen POAG patients and 15 healthy volunteers underwent stimulations with fMRI retinotopic mapping stimulus and contrast-reversing checkerboard patterns stimulus on a Siemens Trio 3.0 T MRI whole-body scanner for functional data collection. Comparisons of V1 fMRI responses between the glaucomatous eyes and the healthy eyes of the patients were carried out using paired samples t-test, while independent samples t-test was used to compare V1 fMRI responses and activations between the healthy eyes of patients and the age-, gender- and side- matched eyes of normal people. Differences of V1 cortical functions and visual functions were analyzed by linear correlation analysis when the glaucomatous and the healthy eyes were simulated individually. Results (1) V1 fMRI responses of the individually stimulated glaucomatous eyes[(1.24±0.72)%]were weaker than those of the healthy eyes[(2.18±0.93)%](t=4.757,P<0.01). Comparisons of V1 fMRI responses between the glaucomatous eyes and matched eyes of normal people, as well as between the healthy eyes of patients and the matched eyes of normal people, were performed respectively: the responses in the glaucomatous eyes[(1.24±0.72)%]were weaker than those in the matched eyes of normal people[(2.01±0.65)%](t=-3.011,P<0.01). There was no statistical difference of the responses between the healthy eyes from patients[(2.18±0.93)%]and the matched eyes of normal people[(1.95±0.75)%](t=0.742,P＞0.05). (2) Differences of V1 cortical functions were negatively correlated with those of visual functions in the individually stimulated glaucomatous and healthy eyes (r=-0.887, P<0.01). (3) The activated area indexes of V1 cortexes in the healthy eyes from patients (0.72±0.12) were lower than those in the matched eyes of normal people (0.85±0.09) (t=-3.801, P<0.01). Conclusion Cortical function impairment was in accordance with visual function impairment in glaucoma. Located and quantified measurement with fMRI retinotopic mapping was a useful method for clinical follow-up and evaluation of functional alteration of glaucomatous visual cortex, and a potentially useful means of studying trans-synaptic degeneration of visual pathways of in vivo glaucoma.

Objective:To explore the effect of high intensity interval training (HIIT) on cardiac rehabilitation in patients after heart transplantation.Methods:According to the search terms, the search was conducted on China National Knowledge Infrastructure, VIP, WanFang Data, China Biology Medicine disc, Web of Science, PubMed, Cochrane Library, Embase, and EBSCO. The search time limit was from the establishment of the database to January 31, 2022. After 2 researchers screened the article, extracted information, and evaluated the quality, a Meta-analysis was conducted using RevMan 5.4 software.Results:According to the inclusion and exclusion criteria, 10 English articles were selected, including 191 patients in the intervention group and 212 patients in the control group, with a total of 403 patients. Meta-analysis showed that during cardiac rehabilitation exercise in patients after heart transplantation, HIIT could improve peak oxygen uptake in cardiopulmonary function exercise testing [ MD=1.98, 95% confidence interval ( CI) (0.55, 3.41), P=0.007], peak heart rate in chronotropic responses [ MD=6.93, 95% CI (2.62, 11.24), P=0.002], and muscle exercise ability [ MD=337.18, 95% CI (12.02, 62.35), P=0.04]. There was no statistically significant difference in systolic blood pressure, diastolic blood pressure, peak systolic blood pressure, peak diastolic blood pressure, resting heart rate and respiratory exchange rate between the two groups ( P>0.05). A subgroup analysis of peak oxygen uptake was conducted based on the intervention period and the start time of rehabilitation exercise after heart transplantation. The results showed that there were statistically significant differences in peak oxygen uptake between the intervention group and the control group when the intervention period was ≤ 12 weeks or the start time was > 6 weeks ( P<0.01) . Conclusions:HIIT effectively improves the peak oxygen uptake, peak heart rate, and muscle exercise activity of patients after heart transplantation. HIIT has a significant impact on peak oxygen uptake when the rehabilitation exercise start time after heart transplantation is > 6 weeks or the intervention period is ≤ 12 weeks.

OBJECTIVETo study the clinical value and operation skills of nasal endoscopy-assisted bulboprostatic anastomosis in the treatment of posterior urethral stricture.

METHODSBetween January 2012 and November 2014, we performed nasal endoscopy-assisted bulboprostatic anastomosis for 12 male patients with posterior urethral stricture. We recorded the operation time, blood loss, exposure of operation visual field, and success rate of anastomosis and summarized the operation skills.

RESULTSEight of the patients experienced first-stage recovery. Two underwent a urethral dilation at 3 months postoperatively, 1 received 10 urethral dilations within 5 months after surgery, and 1 underwent internal urethrotomy after failure in urethral dilation, but all the 4 cases were cured.

CONCLUSIONNasal endoscopy can significantly improve the operation field exposure, elevate the precision, reduce the difficulty, and enhance the efficiency of bulboprostatic anastomosis in the treatment of posterior urethral stricture.

Anastomosis, Surgical ; Endoscopy ; Humans ; Male ; Operative Time ; Postoperative Period ; Urethra ; pathology ; surgery ; Urethral Stricture ; surgery

OBJECTIVETo investigate cause of chronic wounds and related status of patients so as to provide strategy for study and treatment of chronic wounds and establishment of health policy.

METHODSA total of twelve thousand one hundred and sixty-one cases hospitalized in our hospital in 2008 were enrolled in the study. A chronic wound was defined as skin tissue defect which could not heal after one month of treatment. Medical records were thus screened. Then a retrospective study was performed on patients with chronic wounds with analysis of age, gender, injury cause, therapy, and average length of hospital stay. Data were processed with chi-square test and one-way analysis of variance.

RESULTSInvestigation showed: 397 out of 12 161 cases (accounting for 3.3%) were recognized as having chronic wounds. (1) The main causes for chronic wound were burn, diabetes, and pressure ulcer, accounting for 59.9% (238/397), 15.6% (62/397), 10.8% (43/397), respectively. The other causes were operative injury, infection, varicosity, etc. There was statistical difference among the numbers of patients with chronic wounds with regard to various causes of injury (χ(2) = 136.21, P = 0.001). (2) Among patients with chronic wound, the ratio of male and female was 2.0:1.0 with mean age of (44 ± 23) years, and the highest ratio occurred in patients older than 70 years. There was significant difference in the numbers of patients with chronic wound among different age groups (χ(2) = 24.12, P = 0.025). There was statistical difference among the numbers of patients with chronic wound in different age groups with each cause of injury (with χ(2) values from 7.86 to 28.31, P values all below 0.05). (3) All patients with chronic wounds received traditional dressing. In 60.5% (240/397) and 86.4% (343/397) of patients, operative treatment or antibiotics were given. (4) The average length of hospital stay in patients with chronic wound [(38 ± 27) d] was longer as compared with that of all the inpatients in the same period [(15 ± 7) d, F = 22.82, P = 0.012]. There was obvious difference in the average length of hospital stay among patients with chronic wound caused by different reasons (F = 24.06, P = 0.036), in which burn injury resulted in the longest length of hospital stay [(47 ± 27) d].

CONCLUSIONSChronic wounds are mainly caused by diabetes and burn, and characterized by old age and longer length of hospital stay. It is necessary to strengthen translational research and related policy making, so that more rational treatment can be applied for patients with chronic wounds.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Burns ; complications ; Child ; Child, Preschool ; Chronic Disease ; Diabetes Complications ; therapy ; Female ; Humans ; Infant ; Inpatients ; Length of Stay ; Male ; Middle Aged ; Retrospective Studies ; Wound Healing ; Young Adult

To combine fingerprint and drug release rate in vitro, in order to study in vitro release of complex components of Chuanping sustained tablets, compound traditional Chinese medicine preparation. A qualitative determination of the characteristic peaks of the compound preparations were conducted by the fingerprint. The results of the dissolution rate determination under different release conditions showed that the release of three index components (methamphetamine, pseudoephedrine and scopolamine) of Chuanping sustained tablets was less affected by gastrointestinal factors, with similarity factors being more than 80 with unknown component release curves of three major characteristic peaks in the fingerprint. The qualitative determination proved that multiple components of the compound traditional Chinese medicine preparation was dissolved in vitro at similar rates, realizing the balanced release of complex components of the compound traditional Chinese medicine preparation. This study layed a theoretical and experimental basis for quality evaluation for the compound traditional Chinese medicine preparation.
Drugs, Chinese Herbal ; chemistry ; Tablets

OBJECTIVETo optimize the best acupuncture therapy in the treatment of lumbar disc herniation (LDH) and to explore its mechanism in terms of immunology.

METHODSOne hundred and eleven cases were randomized into an acupuncture-moxibustion group, an elongated needle group and a herb-partitioned moxibustion group, 37 cases in each group. In acupuncture-moxibustion group, penetration needling with elongated needle was applied at Dachangshu (BL 25), Guanyuanshu (BL 26) and Huantiao (GB 30), which was followed with herb-partitioned moxibustion at Shenque (CV 8) and Yaoyangguan (GV 3), etc, once a day. Elongated needle and herb-partitioned moxibustion were applied in the elongated needle group and herb-partitioned moxibustion group respectively. The efficacy of each group was observed after 21 treatments and the value of serum immunoglobulin was measured and analyzed before and after treatment.

RESULTSThe effective rate was 97.3% (36/37) in acupuncture-moxibustion group, which was superior to 75.7% (28/37) in the elongated needle group and 73.0% (27/37) in the herb-partitioned moxibustion group (both P<0.05). The visual analogue scale (VAS), straight-leg raising range and motion range of the lumbar vertebra in three groups were all improved after treatment (all P<0.05), which was more significant in the acupuncture-moxibustion group (all P<0.05). The difference of value of serum immunoglobulin before and after treatment was significant in the acupuncture-moxibustion group (P<0.05), which was not in the elongated needle group and the herb-partitioned moxibustion group (both P>0.05).

CONCLUSIONThe therapy of elongated needle combined with herb-partitioned moxibustion achieves superior efficacy in the treatment of LDH as compared with elongated needle or herb-partitioned moxibustion therapy, and it can regulate the humoral immunity system of patients.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

AIM:To observe the changes of autophagy-related indexes during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT) and its effect on apoptosis in human normal hepatocytes.METHODS:LO2 cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS.The expression of glucose-regulated protein 78 (GRP78) , protein kinase R-like endoplasmic reticulum kinase (PERK) , activating transcription factor 4 (ATF4) , C/EBP homologous protein (CHOP) , autophagy-related gene 12 (Atg12) , autophagy-related gene 5 (Atg5) and microtubule-associated protein 1 light chain 3 (LC3) at mRNA and protein levels was determined by real-time PCR and Western blot.The apoptosis was analyzed by flow cytometry.The formation of autophagosomes was observed under transmission electron microscope.After the LO2 cells were pretreated with rapamycin at 400 nmol/L for 1 h and treated with DTT at 2.0mmol/L for 24 h, the effect of rapamycin pretreatment on the apoptosis was analyzed by flow cytometry.RESULTS:After treatment with DTT at 2.0 mmol/L for 6, 12 and 24 h, the mRNA and protein levels of GRP78, PERK, ATF4, CHOP, Atg12, Atg5 and LC3 in the LO2 cells were significantly higher than those in 0 h group (P<0.05).At the same time, the ratio of LC3Ⅱ/LC3Ⅰwas also increased after DTT treatment (P<0.05).Observation under transmission electron microscope showed that autophagosomes were found in the LO2 cells treated with DTT for 6, 12 and 24 h.After DTT treatment for 6, 12 and 24 h, the apoptosis rate of LO2 cells was significantly higher than that in DTT 0 h group, while the apoptosis induced by DTT was significantly decreased after rapamycin pretreatment (P<0.05).CONCLUSION:ERS induces autophagy and rapamycin pretreatment alleviates the apoptosis of LO2 cells to some extent.

The effects of RNAi-mediated gene silencing of LRIG3 expression on cell cycle and survival of human glioma cell line GLI 5 and the possible mechanisms were explored.The plasmids pGenesil2-LRIG3-shRNA l and pGenesil2-LRIG3-shRNA2 were transfected into GL 15 glioma cells respectively by using Metafectine,and the transfected cells that stably suppressed LRIG3 expression were selected by G418.The control cells were transfected with negative control shRNA.The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot.The apoptosis rate and cell cycle were analyzed by flow cytometry.As compared with the negative shRNA-transfected GL15 cells,LRIG3 mRNA expression in GLI5 cells transfected with pGenesil2-LRIG3-shRNAl and pGenesil2-LRIG3-shRNA2 was silenced by 52.4%,63.8%,and LRIG3 protein expression was re-duced by 50.9% and 67.4% respectively.The LRIG3-specific siRNA transfected cells had higher proliferation rate than control cells.Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and the proliferation index significantly (P<0.01).Silencing LRIG3 could inhibit the apoptosis of GLl5 cells (P<0.05).These findings suggest that the siRNA targeting LRIG3 gene shows a dramatic inhibitory effect on RNA transcription and protein expression,then promoting the proliferation of GLI5 cells,arresting GLI5 cells in G2/M phase,and suppressing apoptosis ofGL15 cells.