ObjectiveTo observe the growth of BT-325 human glioma cells after interfering volume-regulated chloride channel ClC-2 gene.MethodsTwo expression recombinant vectors of ClC-2 gene were designed and constructed. The primary plasmid, pSUPER.puro-shRNA, and the two recombinant plasmids, pSUPER.puro-shRNA-ClC-21 and pSUPER.puro-shRNA-ClC-22, were transfected into BT-325 cells by LipofectamineTM2000 (Groups: control, PP1 and PP2, respectively). The mRNA expression of ClC-2 gene was detected with reverse transcription polymerasse chain reaction (RT-PCR), the cellular survival was determined with MTT assay, and the cell cycle was measured with flow cytometry (FCM). ResultsClC-2 mRNA expression and the growth of the cells in PP1 and PP2 groups were significantly lower than that of control group. The cell cycle progression was blocked in G1 phase (PP1 and PP2 vs control,P<0.01). ConclusionThe growth of BT-325 human glioma cells is prevented by knockdown of ClC-2 gene expression, which may be one of the novel targets to inhibit growth of human malignant glioma cells.

Guinea pigs were given orally with a daily dose of 0.5 mg ascorbic acid per 100 g body weight 20 days before and 4 days after a whole-body r-irradiation and then injected subcutaneously with 10 mg ascorbic acid at 24 hours intervals. During the injection period, the daily urinary output of total ascorbic acid in the animals irradiated with 500 or 1000 r was significantly higher than that in controls, but the difference of 24 hours excretions of total ascorbic acid between the group irradiated with 150 or 250 r and the control group was not statistically significant.As shown in the urinary output of dehydroascorbic acid and diketo- gulonic acid estimated by our modified Roe's method, r-irradiation has no effect on its excretion in guinea pigs and rats.The levels of ascorbic acid in plasma, spleen and adrenals were significantly lower in the irradiated group than in the control group, but might increase with the increase of the injected dose of ascorbic acid.

Objective:To investigate the effects of different analgesic methods of hydromorphone on analgesic efficacy and sleep quality in patients with refractory cancer pain.Methods:Sixty patients with refractory cancer pain who received three-step analgesic treatment in Quzhou People's Hospital from August 2018 to December 2019 and acquired poor analgesic effects were included in this study. They were randomly assigned to undergo either an intravenous patient-controlled analgesia with hydromorphone (HV group, n = 30) or an intrathecal patient-controlled analgesia with hydromorphone (HI group, n = 30) for 10 consecutive days. The analgesic efficacy in each group was evaluated using the numerical rating scale (NRS) before and 2, 4, 24, 48 hours, and 10 days after administration. The frequency of breakthrough pain (BTP) at each time point was recorded. The sedation effect of medication was evaluated using the Ramsay score. The sleep quality of patients was evaluated using the Pittsburgh sleep quality index (PSQI). The activities of CD 3+, CD 4+, and CD 4+/CD 8+ lymphocyte subsets were measured by flow cytometry at different time points. The adverse reactions within 10 days after treatment were observed and recorded. Results:Before and at each time point after treatment, there were no significant differences in NRS score, the frequency of BTP, Ramsay score, and PSQI score between the two groups (NRS score: t = 0.45, 0.91, 0.52, 1.19, 0.97, 1.92, all P > 0.05; frequency of BTP: t = 0.34, 1.88, 0.86, 1.71, 1.22, 0.76, all P > 0.05; Ramsay score: t = 0.56, 0.46, 0.63, 0.22, 0.99, 0.14, all P > 0.05; PSQI: t = 0.86, 1.25, 1.46, 1.05, 0.57, 1.93, all P > 0.05). At each time point after treatment, the activities of CD 3+, CD 4+, and CD 4+/CD 8+ cells increased in each group, and the activities of CD 3+, CD 4+, and CD 4+/CD 8+ cells in the HI group were significantly higher than those in the HV group (CD 3+: t = 3.72, 3.12, 2.85, 3.13, 2.44, all P < 0.05; CD 4+: t = 3.62, 2.45, 3.31, 3.19, 2.70; all P > 0.05; CD 4+/CD 8+: t = 3.10, 2.74, 2.83, 3.24, 3.41, all P < 0.05). The total incidence of adverse reactions was slightly, but not significantly, lower in the HI group than the HV group [14.00% (7/30) vs. 26.00% (13/30), χ2 = 2.70, P = 0.100]. Conclusion:Compared with intravenous administration of hydromorphone, intrathecal administration of hydromorphone can better effectively relieve pain, decrease the frequency of BTP, improve sleep quality, has a good sedative effect, improve immune function, and has fewer adverse reactions.

BACKGROUNDGastroesophageal reflux disease with extra-esophageal symptoms, especially those with respiratory distress was attracting more and more attention. The related mechanisms were still in controversy. The purpose of the work was to explore airway inflammation triggered by gastroesophageal reflux.

METHODSSixteen Sprague-Dawley rats were used as study group and 9 as control. In the study group, a plastic extender with a trumpet-shaped distal end was inserted into the lower esophagus to dilate the cardia, the pylorus was ligated. One ml of 0.1 mol/L hydrochloric acid was injected into the stomach. While a simple laparotomy was performed for control animals. All animals from two groups were sacrificed 24 hours after operation. Then tracheotomy was carried and the bronchoalveolar lavage fluid was collected in all animals. Cells in the fluid were counted and levels of interleukin (IL)-5, -6, -8 in it were measured.

RESULTSCompared with control group, the study group presented a neutrophil pattern of airway inflammation and an elevated concentration of IL-5, -6, -8 with no significant difference regarding eosinophil count.

CONCLUSIONThe gastroesophageal reflux-triggered airway inflammation is characterized by a neutrophilic airway inflammation which differed from that caused by asthma, and enhanced levels of IL-5, -6 and -8, which are similar to that caused by asthma.

Animals ; Asthma ; etiology ; Bronchoalveolar Lavage Fluid ; immunology ; Disease Models, Animal ; Female ; Gastroesophageal Reflux ; complications ; Inflammation ; etiology ; Interleukin-5 ; analysis ; Interleukin-6 ; analysis ; Interleukin-8 ; analysis ; Male ; Rats ; Rats, Sprague-Dawley

Calcifying tendinopathy is a tendon disorder with calcium deposits in the mid-substance presented with chronic activity-related pain, tenderness, local edema and various degrees of incapacitation. Most of current treatments are neither effective nor evidence-based because its underlying pathogenesis is poorly understood and treatment is usually symptomatic. Understanding the pathogenesis of calcifying tendinopathy is essential for its effective evidence-based management. One of the key histopathological features of calcifying tendinopathy is the presence of chondrocyte phenotype which surrounds the calcific deposits, suggesting that the formation of calcific deposits was cell-mediated. Although the origin of cells participating in the formation of chondrocyte phenotype and ossification is still unknown, many evidences have suggested that erroneous tendon cell differentiation is involved in the process. Recent studies have shown the presence of stem cells with self-renewal and multi-differentiation potential in human, horse, mouse and rat tendon tissues. We hypothesized that the erroneous differentiation of tendon-derived stem cells (TDSCs) to chondrocytes or osteoblasts leads to chondrometaplasia and ossification and hence weaker tendon, failed healing and pain, in calcifying tendinopathy. We present a hypothetical model on the pathogenesis and evidences to support this hypothesis. Understanding the key role of TDSCs in the pathogenesis of calcifying tendinopathy and the mechanisms contributing to their erroneous differentiation would provide new opportunities for the management of calcifying tendinopathy. The re-direction of the differentiation of resident TDSCs to tenogenic or supplementation of MSCs programmed for tenogenic differentiation may be enticing targets for the management of calcifying tendinopathy in the future.
Animals ; Cell Differentiation ; physiology ; Humans ; Mice ; Rats ; Stem Cells ; pathology ; Tendinopathy ; etiology ; pathology ; Tendons ; pathology

This study was aimed to investigate the expression of blood Th17 and CD4(+) CD25(+) regulatory T cells (Treg) in the patients with aplastic anemia (AA). Forty-five patients with AA were enrolled into this study, and were divided into mild aplastic anemia (MAA) group (n = 25) and severe aplastic anemia group (SAA) (n = 20), blood cell count was recorded. 15 healthy donors were enrolled as control. Proportions of blood Th17 and CD4(+) CD25(+) Treg cells were determined by flow cytometry. The serum levels of IL-17, IFN-γ and TNF-α, as well as their concentrations in culture supernatant of phytohemagglutinin (PHA) -stimulated peripheral blood mononuclear cells, were measured by ELISA. The results showed that the proportions of blood Th17 cells and concentration of blood serum IL-17 and IFN-γ increased in patients with SAA, compared with MAA and normal controls, but CD4(+) CD25(+) Foxp3(+) Treg cells obviously decreased in patients with SAA. The concentrations of IL-17 and IFN-γ significantly increased in culture supernatant of SAA group. Hemoglobin level in the patients with AA negatively correlated with the population of Th17 cells and serum IL-17 level, whereas positively correlated with the expression of CD4(+) CD25(+)Treg cells. It is concluded that the increased response of Th17 cells and deficiency of CD4(+) CD25(+) Treg cells present in severe aplastic anemia. The severity of anemia may be related with the imbalance between Th17 and CD4(+) CD25(+)Treg cell response.
Adult ; Aged ; Anemia, Aplastic ; blood ; metabolism ; Case-Control Studies ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-17 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; metabolism ; Th17 Cells ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

Aged ; Animals ; Asphyxia ; etiology ; Asthma ; physiopathology ; Catheter Ablation ; Female ; Fundoplication ; Gastroesophageal Reflux ; complications ; surgery ; Humans ; Laryngismus ; etiology ; Male ; Middle Aged ; Rats ; Rats, Sprague-Dawley ; Respiration Disorders ; etiology ; physiopathology ; Treatment Outcome

OBJECTIVETo investigate the changes in transforming growth factor beta 1 (TGF-beta1)/Smads signaling pathway in rats with chemical hepatocarcinogenesis.

METHODSFresh diethylnitrosamine (DENA) solution was administered in SD rats to induce hepatocellular carcinoma (HCC). The protein expressions of TGF-beta1, phosphorylated Smad2, Smad4 and Smad7 were detected in these rats with immunohistochemistry, and the mRNA expression of Smad4 was evaluated with RT-PCR.

RESULTSCirrhotic nodules occurred in the rats 8 weeks after DENA treatment, and HCC nodules were found 16 weeks after the treatment. In the normal liver tissue, very low levels of TGF-beta1 and Smad4 expressions, low Smad7 expression and high phosphorylated Smad2 expression were detected. The development of liver cirrhosis was accompanied by increased expressions of TGF-beta1, Smad4 and Smad7 but at 8 weeks after DENA treatment, the expression of phosphorylated Smad2 was significantly decreased, followed then by gradual increment till nearly the normal level. Twenty-two weeks after DENA treatment, Smad4 expression in liver tissue decreased markedly as compared with the levels at 8 and 16 weeks. The expressions of Smad4 and phosphorylated Smad2 in the HCC tissue was significantly lower than those in normal liver tissue.

CONCLUSIONHepatocarcinogenesis involves very complex mechanisms, can can be related partially to the decreased Smad4 and phosphorylated Smad2 expression and TGFbeta1 and Smad7 overexpression in advanced stage of liver cirrhosis.

Animals ; Diethylnitrosamine ; Liver Neoplasms, Experimental ; chemically induced ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad2 Protein ; metabolism ; Smad4 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

BACKGROUNDTo confirm if Puumala like viruses exist in China.

METHODSRNA was extracted from lungs of bank voles captured in the Northeast China, partial S segments genome of Puumala viruses were amplified and sequenced.

RESULTS926 bp cDNA of S segments of Puumala like virus was amplified and sequenced. The phylogenetic analysis revealed that the Puumala like viruses found in China were most close to that found in Far East region of Russia.

CONCLUSIONSPuumala like virus does exist in Northeast China, and the nucleotides sequence of the viruses have high homolog to Puumala viruses found in Russia.

Animals ; China ; DNA, Viral ; analysis ; Hemorrhagic Fever with Renal Syndrome ; virology ; Lung ; virology ; Mice ; Phylogeny ; Puumala virus ; isolation & purification ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo observe the efficacy of Jianbu Tongluo Xunzheng Liquid (JTXL) in treating knee osteoarthritis (KOA), and to explore the correlation between changes of infrapatellar fat pad scanned by CT and the efficacy.

METHODSTotally 105 KOA outpatients were randomly assigned to three groups, i.e., the treatment group, the control group, and the combination group, 35 in each group. Patients in the treatment group were fumigated by JTXL, 30 min each time, once daily, 10 times as a course of treatment, 3 courses in total. Those in the control group received intra-articular injection of Sodium Hyaluronate Injection (SHI), 3 mL each time, once per 6 days, 5 times in total. Those in the combination group were treated by fumigation of JTXL + intra-articular injection of SHI in the same way as the aforesaid two groups. All patients were treated for 30 days. Their clinical efficacy and changes of infrapatellar fat pad scanned by CT were observed, and their correlation was analyzed.

RESULTSThe total effective rate was 88.57% in the combination group, better than that of the control group (74.29%) and the treatment group (80.00%; both P < 0.05). Besides, the score for knee joint functions at Hospital for Special Surgery (HSS) was better in the combination group than the other two groups (P < 0.05). The anteroposterior diameter, exterior-interior diameter, the superior-inferior diameter were shortened, and the density decreased in the treatment group and the combination group (P < 0.05). Besides, they were superior to those of the control group (P < 0.05).

CONCLUSIONSChanges of infrapatellar fat pad scanned by CT only existed in the combination group and the treatment group, indicating changes of CT scanning was only correlated with effect on changing physicochemical properties of infrapatellar fat pad. Treatment by Chinese medicine could omnipotently and balanced regulate functions and structures of every tissue. Therefore, CT could be taken as a better method for clinical efficacy observation by Chinese medicine.

Adipose Tissue ; diagnostic imaging ; Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; diagnostic imaging ; drug therapy ; Patella ; diagnostic imaging ; Phytotherapy ; Tomography, X-Ray Computed