Background and purpose:It is still a great challenge to clarify the signal molecules that mediate the communication between cancer-associated fibroblasts(CAFs)and tumor cells.These signal molecules are very important for cancer metastasis.The purpose of this study was to explore the communication mechanism of pleckstrin-2/miR-196a signal axis mediated by lung cancer cells in tumor microenvironment.Methods:Human lung adenocarcinoma cell line H1299 and human embryonic lung cell MRC-5 were selected as the research objects.H1299 cells were transfected with lentivirus(PLEK2)expressing PLEK2 and Vector control,and exosomes(Vector_exo,PLEK2_exo)were isolated after 24 h of transfection.MRC-5 cells were transfected with miR-196a mimetic or inhibitor.The expressions of PLEK2 and epithelial-mesenchymal transition(EMT)-related proteins were analyzed by Western blot.The expression of miR-196a was analyzed by polymerase chain reaction(PCR),and the metastasis and invasion ability of cells were determined by transwell assay.Six female BALB/c-nu mice were randomly divided into Vector group and PLEK2 group,with 3 mice in each group.Mice in each group were injected with H1299 cells transfected with Vector or PLEK2 through the tail vein.After 4 weeks,lung tissue was taken out for H-E staining and immunohistochemical staining to analyze the expression of α-smooth muscle actin(α-SMA).All animal experiments were approved by the ethics committee of First Hospital of Changsha City(Changsha Hospital,Xiangya School of Medicine,Central South University)(ethics number:EI-2021-103).Results:Compared with the Vector group,the number of pulmonary metastatic nodules and the expression of α-SMA in metastatic cancer in PLEK2 group increased significantly(P＜0.001).Compared with Vector group,the expression level of miR-196a in H1229 cells in PLEK2 group increased significantly(P＜0.05),and the expression level of miR-196a was significantly higher in PLEK2_exo than in Vector_exo(P＜0.05).Compared with Vector_exo group,the expression levels of miR-196a,α-SMA and fibroblast activation protein(FAP)in MRC-5 cells in PLEK2_exo group increased significantly(P＜0.05).Compared with the negative control(NC),the expression levels of α-SMA and FAP in MRC-5 cells transfected with miR-196a increased significantly(P＜0.05).On the contrary,by transfection with miR-196a inhibitors(si-miR-196a#1 and si-miR-196a#),the expression levels of α-SMA and FAP were significantly inhibited(P＜0.05).Compared with NC-CM group,the number of metastatic cells,invasive cells and the expression of vimentin in miR-196a-CM group increased significantly(P＜0.001),and the expression of E-cadherin decreased significantly(P＜0.001).In addition,compared with Vector_exo-CM group,PLEK2_exo-CM group had significant increase in number of metastatic and invasive cells and the expression of vimentin(P＜0.01),and significant decrease in the expression of E-cadherin(P＜0.001).Conclusion:Upregulation of PLEK2 can enhance the level of exosomes miR-196a derived from lung cancer cells,thereby promoting the activation of CAFs.The activated CAFs can further enhance the invasive ability of lung cancer cells.

The clinical data, laboratory test, and gene mutations were collected from a family with pseudohypoaldosteronism type II(PHA2). The proband, aged 1 year and 7 months, presented with hyperkalemia(6.69 mmol/L; reference range 3.5-5.3 mmol/L), blood pressure of 110/68 mmHg(normal<106/61 mmHg, 1 mmHg=0.133 kPa), blood chloride of 111.5 mmol/L(reference 99-110 mmol/L), blood HCO 3- of 17.1 mmol/L(reference 22-29 mmol/L), estimated glomerular filtration rate of 128.5 mL·min -1·(1.73 m 2) -1[>90 mL·min -1·(1.73 m 2) -1], and blood renin concentration of 0.30 μIU/mL(reference 4.2-45.6 μIU/mL). The mother and maternal grandfather also exhibited normal renal function with hyperkalemia, hypertension, hyperchloremia, metabolic acidosis, and low renin. Genetic testing revealed a heterozygous missense mutation(c.1685A>G, p. E562G) in exon 7 of the no-lysine kinase 4(WNK4) gene. Treatment with hydrochlorothiazide was effective. Literature review comparing this E562G pedigree with other WNK4 variants suggested clinical heterogeneity of WNK4 mutations. For unexplained hyperkalemia, especially with concurrent hypertension, PHA2 should be considered early for genetic screening to prevent misdiagnosis or delayed diagnosis.

Objective:To explore the effect of standardized operation procedure of prone position placement in patients undergoing posterior vertebral pedicle screw system internal fixation.Methods:From May 2019 to May 2021, 98 patients with spinal stenosis who were treated with posterior vertebral pedicle screw system internal fixation in Wuxi Second People's Hospital were selected as the study subject by convenience sampling. The patients were randomly divided into the control group and the observation group with 49 cases in each group. The control group was given routine nursing, and the observation group received standardized operation procedure of prone position placement. The time needed to complete the position placement, the time of position recovery, the incidence of pipeline accidents and the total incidence of pressure injuries were compared between the two groups.Results:After the intervention, the time of position placement and recovery in the observation group were shorter than those in the control group, with statistical differences ( P<0.05) . The incidence of pipeline accidents in the observation group was 2.05%, lower than that in the control group 12.24%; the total incidence of stress injury in the observation group was 6.12%, lower than 18.37% in the control group; but there was no significant difference between the two groups ( P>0.05) . Conclusions:Standardized operation procedure of prone position placement can clarify the division of labor, shorten the prone position placement and recovery time of patients undergoing posterior vertebral pedicle screw system internal fixation, to some extent, it can reduce the risk of pipeline accidents and pressure injuries, and provide safe surgical experience for patients.

Root canal obturation is conducted by using filling materials to tightly seal the root canal system after the procedure of preparation in order to control infection and promote periapical healing. The quality of root canal obturation is one of the essential factors affecting the prognosis of root canal treatment. Qualified root canal filling is defined as a homogeneous radiographic apical filling within the cemento-dentine junction with neither overfilling nor underfilling. This review elucidates the long-term outcome of root canal overfilling and its causes, the influence of apical overfilling on adjacent structures and the prevention and management of overfilling, so as to help the clinicians achieving a better outcome of root canal treatment and obtaining an optimal long-term prognosis.

Objective To analyze the relationship between pain sensation, emotion and recognition in three dimensions. Methods By using questionnaires which contained general information questionnaire, Cancer Pain Questionnaire, Self-reporting Inventory (SCL-90), Pain Beliefs and Perceptions Inventory (PBPI) to investigate pain sensation, emotion and recognition of 46 patients with cancer pain. Results There were 13(28.3%) cases sufferd from mild pain,17 (37.0%) cases were moderate pain, 16 (34.8% )cases were severe pain.As to the result of SCL- 90,patients showed obvious symptom in somatization, depression, anxiety and hostility.They holded deep belief of that pain was very mysterious. There was a significant correlation between pain severity and depression(rs=0.377) , anxiety(rs=0.388) on the condition that confidence level was 0.01;there was also a significant correlation between pain degree and interpersonal sensitivity(rs=0.308), hostility(rs=0.320) on the condition that confidence level was 0.05. As to pain beliefs, pain degree had a significant correlation with it in the dimension of pain as mystery (rs=0.529) and pain was persistent(rs=0.680) on the condition that confidence level was 0.01. Conclusions The survey shows a positive correlation between pain severity,emotion of pain(such as anxiety,depression, hostility and interpersonal sensitivity)and beliefs about pain as mystery or permanent.

Objective To investigate whether the effects of rapamycin on promoting oral squamous cancer cell apoptosis is related with inhibiting TLR4 expression and inflammatory factor IL-6 and PGE2 expression.Methods Human oral squamous carcinoma SCC-15 cell line was cultured and interfered by rapamycin.Then the activity of SCC-15 cells was detected by CCK 8,the SCC-15 cells apoptosis was detected by the Hochest33342/PI double staining,the invasion ability was determined by the transwell method.The TLR4 protein expression level was detected by Western blot and IL-6 and PGE2 expressions were detected by ELISA.Results Rapamycin could promote cell apoptosis,the invasion ability of SCC-15 cells was significantly decreased.After rapamycin intervention,the TLR4 protein expression was decreased and expression levels of IL-6 and PGE2 were reduced,showing statistical difference as compared with the negative group (P＜0.05).Conclusion Rapamycin promotes oral squamous cancer SCC-15 cell apoptosis,which may be related with inhibiting TLR4,IL-6 and PGE2 expression.

ObjectiveTo investigate the effect of therapeutic endoscopic retrograde cholangiopancreatography (ERCP) on liver function and treatment outcome in patients with pancreaticobiliary diseases. MethodsA total of 397 patients with pancreaticobiliary diseases who underwent ERCP with an electronic duodenoscope in Beijing Chuiyangliu Hospital from January 2005 to July 2015, were enrolled. According to the type of disease, they were divided into benign obstruction group (109 patients), malignant obstruction group (47 patients), pancreatitis group (27 patients), foreign body obstruction group (127 patients), simple cholangitis group (19 patients), biliary fistula group (15 patients), juxta-ampullary duodenal diverticula group (29 patients), and liver transplantation group (24 patients). Liver function parameters including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), and total bilirubin (TBil) were measured at 1 day before ERCP and at 4 weeks after ERCP. The therapeutic effect was evaluated at 1 month after ERCP. The paired t-test was used for comparison of continuous data before and after treatment within one group, an analysis of variance was used for comparison between multiple groups, and the chi-square test was used for comparison of categorical data between groups. ResultsAfter ERCP, all the patients showed significant improvements in ALT, AST, ALP, GGT, and TBil, suggesting that ERCP significantly improved the subjects′ liver function (all P＜0.05). The evaluation of therapeutic effect showed that after ERCP, all the groups showed significant remission of symptoms; all the patients in the simple cholangitis group were cured, while those in the liver transplantation group and malignant obstruction group only achieved certain improvements. ConclusionTherapeutic ERCP can significantly improve the patients′ liver function. However, as for patients with liver transplantation and pancreaticobiliary tumors, laparoscopic surgery should be performed in combination with ERCP.

Objective To prepare the infected Oncomelania hupensis by artificial method for the research on the activity, vaccine, and genetic variation of Schistosoma Japonicum (S. Japonicum).Methods The mature eggs of S. Japonicum were collected by Nylon silk method and the miracidia were incubated under appropriate conditions. Negative snails were infected with miracidia in different proportion by means of individual or collective infection to seek the best method and proportion of infection between miracidia and snails. Infected snails were divided into 12 groups in total. Ⅰ-Ⅵ groups were for individual infection and Ⅶ-Ⅻ groups were for collective infection. There were 200 snails in each group. The infection ratios between snails and miracidia in Group Ⅰ-Ⅵ or screened, numbered, and reared singly. The amount of cercariae was calculated once every 10 days until the infected snails died. Then cercariae shedding quantity, infection quantity, and mortality of infected snails in every group were compared to find the best infection method and the best infection proportion between miracidia and snails. The cercariae were collected from the first generation of infected snails and were used to infect experimental animals. The mature eggs of S. Japonicum were saved from the infected experimental animals and incubated to get miracidia. The snails were artificially infected by miracidium to get the second generation of infected snails. The developmental rates of adult worms, the egg density in fecal and liver were compared between artificially and naturally infected snails. Results In individual infection GroupⅠ-Ⅵ,the average infection value of snails were 0±0,22.7±4.2,31.7±4.5,53.0±5.3,39.3±5.9,32.7±4.7,the average fatality of snails were 21.7±3.1,25.0±3.6,31.3±4.9,44.7±6.5,78.3±9.5,89.7±13.6, and the average value of cercariae shedding from infected snails were 0.0±0.0,308.0±96.6,428.1±146.2,527.0±171.1,571.4±148.9,602.9±356.3, respectively. In collective infection Group Ⅶ-Ⅻ,the average infection value of snails were 0±0,12.3±2.5,18.7±4.7,28.3±4.2,33.3±4.7,29.3±5.5,and the average fatality of snails were 22.7±3.8,23.7±4.5,28.3±5.5,47.0±9.5,75.7±8.5,86.3±12.2, and the average value of cercariae shedding from infected snails were 0±0,244.5±57.3,292.3±74.8,347.1±100.8,477.2±142.1,447.3±161.4, respectively. The second generation of artificially infected snails was obtained successfully. The average infection rate and fatality rate for the second generation of artificially infected snails were 24.65% and 24.50%, both of which were not obviously different from that of the first generation of artificially infected snails (P>0.05). In the animal experiment, the worm growth rate for the naturally infected snails, the first or second generation of artificially infected snails were 68.50%,73.50% or 71.00%. There was no obvious difference among them (P>0.05). The fecal (or liver) eggs per gram for the naturally infected snails, the first or the second generation of artificially infected snails were 1 503±269,1 683±233, or 1 541±117 (or 6 641±1 819,6 272±1 419, or 7 263±1 643). There was no significant difference among the 3 groups (P>0.05). Conclusion Infected snails can be obtained through the artificial method by using S. Japonicum miracidia to infect snails. Individual infection has the advantage over collective infection. The optimal proportion of infection between first and the second generation of artificially infected snails in the average of cercariae shedding, infection, and fatality average of snails. There was no significant difference between artificially and naturally infected snails in the developmental rate of adult worms, fecal and liver eggs per gram.

Objective To study the effect of magnetic stimulation on the expression of B cell lymphoma/leukemia gene 2 ( Bcl-2 ) and caspase-3 genes, and the apoptosis of neurons in rats with spinal cord injury (SCI).Methods Sixty rats were randomly divided into a magnetic stimulation group, a model group and a sham-operation group. An SCI model was established in the magnetic stimulation and model groups. The magnetic stimulation was applied at the 6th, 12th, 24th and 72nd hour after the operation to the rats in the magnetic stimulation group, and sham magnetic stimulation was given to the model group and sham-operation group rats at the same time points. Two hours after treatment, 5 rats of each group were sacrificed and their injured spinal cords were sectioned. The gene expressions were detected using immunohistochemical techniques, and apoptosis of neurons was observed by the TUNEL method. Results Few apoptotic cells were found in the sham-operation group, but more were found in the model group. Apoptotic cells in the magnetic stimulation group were significantly fewer than in the model group. The expression of both Bcl-2 and caspase-3 in the magnetic stimulation and model groups was significantly higher than in the sham-operation group at the different time points. Expression of Bcl-2 in the magnetic stimulation group was significantly higher than in the model group, but expression of caspase-3 in the magnetic stimulation group was significantly lower than in the model group. Conclusions Magnetic stimulation up-regulates the expression of Bcl-2 genes and down-regulates the expression of caspase-3 in injured neurons. Magnetic stimulation might have protective and rehabilitative effects after human SCI.

ObjectiveTo investigate the effects of hyperuricemia on motor function, activities of daily living (ADL) in patients with cerebral infarction. Methods120 patients with cerebral infarction were divided into control group (n=60) and hyperuricemia group (n=60). Two groups accepted the similar training of equal time and intensity, and were assessed with Fugl-Meyer Assessment (FMA) and Modified Barthel Index (MBI). ResultsAfter 4 weeks training, the increase of score of FMA and MBI in the hyperuricemia group was obviously lower than in the control group (P＜0.01). ConclusionHyperuricemia could adversely affect the discovery of motor function and ADL in patients with cerebral infarction who accepted rehabilitation.