Objective To investigate the effect of combination of rope-assisted proprioceptive neuromuscular facilitation(PNF)training and rope-assisted brain-computer interface(BCI)training on upper limb function in stroke patients with hemiplegia. Methods From March,2022 to February,2023,96 inpatients with stroke hemiplegia from the Second Affiliated Hospital of Guangxi Medical University were randomly divided into conventional group(n=32),PNF group(n=32)and combined group(n=32).All the groups received routine rehabilitation treatment.The conventional group re-ceived upper limb PNF training,the PNF group received upper limb rope-assisted PNF training,and the com-bined group received both upper limb rope-assisted PNF training and upper limb rope-assisted BCI training,for four weeks.They were assessed with Functiongal Test for the Hemiplegic Upper Extremity-Hong Kong version(FTHUE-HK),Fugl-Meyer Assessment-Upper Extremities(FMA-UE)and modified Barthel Index(MBI)before and after treatment. Results The intra-group effect(F>341.219,P<0.001),inter-group effect(F>21.705,P<0.001)and interaction effect(F>3.171,P<0.05)were significant in the scores of FTHUE-HK and MBI.The intra-group effect(F=520.472,P<0.001)and inter-group effect(F=41.939,P<0.001)were significant in the scores of FMA-UE,and the interaction effect was not(P>0.05).After treatment,the FTHUE-HK,FMA-UE and MBI scores were the best in the combined group(P<0.05). Conclusion The combination of rope-assisted PNF training with rope-assisted BCI device training could further improve the motor function of the upper limbs in stroke patients with hemiplegia,and enhance their activities of daily liv-ing.

ObjectiveTo observe the effect of Youguiwan on the leptin/Janus kinase 2 （JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway in the lung tissue of the rat model of chronic obstructive pulmonary disease （COPD） due to kidney-Yang deficiency. MethodForty rats were modeled for COPD with the syndrome of kidney-Yang deficiency by intratracheal instillation of lipopolysaccharide on day 1 and day 14 and continuous fumigation for 6 weeks， during which hydrocortisone was injected intramuscularly at an interval of 3 days. The modeled rats were randomized into model， high- （11.7 g·kg^-1）， medium- （5.85 g·kg^-1）， and low-dose （2.93 g·kg^-1） Youguiwan， and aminophylline （0.054 g·kg^-1） group. In addition， 8 SD rats were set as the blank group. After the completion of modeling， the rats in each group were administrated with the corresponding drug by gavage for 28 consecutive days. After the last administration， samples were collected. A lung function analyzer was used to evaluate the lung function of rats. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-17A （IL-17A）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the bronchoalveolar lavage fluid （BALF）. Hematoxylin-eosin staining was employed to observe the pathological changes in the lung tissue， and Masson staining was employed to observe the deposition of blue collagen fibers around bronchi in the lung tissue and calculate the inflammation score. The immunofluorescence assay was employed to measure the protein content of collagen type Ⅰ （ColⅠ） and α-smooth muscle actin （α-SMA） in the bronchi. The protein and mRNA levels of leptin， IL-17A， JAK2， and STAT3 in the lung tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction， respectively. ResultCompared with the blank group， the model group showed decreased lung function （P<0.01）， elevated levels of IL-6， IL-17A， and TNF-α in the BALF （P<0.01）， and increased lung inflammation score， deposition of subcutaneous collagen fibers in the airway， and ColⅠ and α-SMA proteins （P<0.01）. Furthermore， the modeling up-regulated the proteins and mRNA levels of leptin， IL-17A， JAK2， and STAT3 in the lung tissue （P<0.01） and enhanced the phosphorylation of JAK2 and STAT3 （P<0.01）. Compared with the model group， high- and medium-dose Youguiwan improved the lung function， decreased the inflammation score， reduced collagen fiber deposition and ColⅠ and α-SMA proteins， lowered the levels of IL-6， IL-17A， and TNF-α in the BALF， down-regulated the mRNA and protein levels of leptin， JAK2， STAT3， and IL-17A， and weakened the phosphorylation of JAK2 and STAT3 （P<0.05， P<0.01）. The aminophylline group had higher IL-17A and TNF-α levels than the high-dose Youguiwan group， lower IL-17A level than the medium and low-dose Youguiwan groups， and lower TNF-α level than the low-dose Youguiwan group. Compared with the aminophylline group， the high- and medium-dose Youguiwan groups showed reduced deposition of collagen fibers and protein levels of ColⅠ and α-SMA around the bronchi in the lung tissue （P<0.05， P<0.01）， decreased inflammation score， and down-regulated protein and mRNA levels of leptin， JAK2， STAT3， and IL-17A in the lung tissue. ConclusionYouguiwan can prevent airway remodeling by inhibiting IL-17A to reduce inflammation and collagen deposition in COPD rats， which may be related to the inhibition of the leptin/JAK2/STAT3 signaling pathway.

ObjectiveTo investigate the effect of combination of proprioceptive neuromuscular facilitation (PNF) rope training and repetitive transcranial magnetic stimulation (rTMS) on upper limb motor function of stroke patients with hemiplegia. MethodsFrom March, 2021 to February, 2022, 90 stroke inpatients with hemiplegia in the Department of Rehabilitation Medicine, the Second Affiliated Hospital of Guangxi Medical University were divided into control group (n = 30), rope group (n = 30) and combination group (n = 30) randomly. All the groups received routine rehabilitation, while the control group received routine PNF training, the rope group received PNF rope training, and the combination group received PNF rope training and rTMS, for four weeks. They were assessed with Functional Test for the Hemiplegic Upper Extremity-Hong Kong version (FTHUE-HK), Fugl-Meyer Assessment-Upper Extremities (FMA-UE) and modified Barthel Index (MBI) before and after treatment. ResultsThe scores of FTHUE-HK, FMA-UE and MBI increased in all the three groups after treatment (t > 2.167, P < 0.05), and they were more in the combination group than in the rope group (P < 0.05), and in the rope group than in the control group (P < 0.05). ConclusionThe combination of PNF rope training and rTMS is more effective on motor function of upper limbs of stroke patients with hemiplegia than PNF training alone.

Objective:To evaluate the efficacy and safety of ultrasound-guided high intensity focused ultrasound(HIFU)on pain intensity,pain sensation and overall survival in patients with advanced pancreatic cancer. Methods:Clinical data of advanced pancreatic cancer patients treated by HIFU were collected from the patients enrolled during August 2020 to September 2022 at the second department for oncology of Yueyang Hospital of Integrated Chinese and Western Medicine affiliated to Shanghai University of Traditional Chinese Medicine.In this study,SPSS 26.0 software was used for the statistical analysis of NRS score and BPI score.The Kaplan-Meier survival analysis method was applied to calculate the median overall survival(OS)and then the survival curve was drawn.At the same time,the incidence of related adverse reactions during and after HIFU treatment was counted. Results:(1)Among the 45 patients,30 patients received HIFU combined with chemotherapy,and the other 15 patients only received HIFU.(2)Among the 45 patients,32 patients had pain relief after HIFU treatment,and the NRS score kept decreased across 1 week,2 weeks,3 weeks and 1 month after HIFU treatment(P＜0.05).The pain sensation score of BPI scale also decreased correspondingly,and the difference was statistically significant(P＜0.05).(3)The median OS of 45 patients was 11.1 months(95％Cl:9.30-1 2.90),of which 30 patients treated with HIFU combined chemotherapy had a median OS of 12.4 months(95％Cl:9.1 8-15.62),and 15 patients treated with HIFU only had a median OS of 4.6 months(95％Cl:1.11-8.10).(4)No serious adverse events were observed in all patients during and after HIFU treatment.Only 5 patients had asymptomatic mild elevation of blood amylase,and the incidence of mild adverse reactions was 11.1％. Conclusion:HIFU can effectively relieve pain and prolong the median survival time in patients with advanced pancreatic cancer.

Objective:To select the relevant evidence of percutaneous pericardial drainage tube nursing and summarize the best evidence.Methods:Evidence-based questions were established based on PIPOST model. BMJ Best Clinical Practice, China Biology Medicine disc (CBM) , UpTodate, Cochrane Library, Joanna Briggs Institute Evidence-based Health Care Database, China Guide Network, British Guide Network, National Guide Line Clearing House (NGC) , PubMed, EMbase, Evidence-based Medicine (EBM) , Registered Nurses' Association of Ontario (RNAO) , The European Society of Cardiology (ESC) , American Heart Association (AHA) , Chinese Journal Full-text Database and Wanfang Database were conducted computer retrieval. The search time was from the establishment of the database to December 31, 2020. Two researchers respectively evaluated the quality of the included literature and extracted data and summarized and summarized the evidence that met the standards.Results:Finally, 12 articles were included, including 1 evidence summary, 2 systematic reviews, 1 systematic assesment, 2 guidelines, 1 expert consensus and 5 case series studies. Finally, 11 pieces of evidence were formed, including 6 themes such as drainage tube selection, puncture wound nursing, drainage flow control, flushing and sealing of the tube, observation and recording points, extubation indications and care.Conclusions:This study summarizes the best evidence for percutaneous pericardial drainage tube nursing, which provides evidence-based basis for improving the quality of percutaneous pericardial drainage tube care.

Objective:To study the effect of COVID-19 on pregnancy outcomes and neonatal prognosis in Hubei Province.Method:s A retrospective comparison of the pregnancy outcomes was done between 16 women with COVID-19 and 45 women without COVID-19. Also, the results of laboratory tests, imaging examinations, and the 2019 novel coronavirus (2019-nCoV) nucleic acid test were performed in 10 cases of neonatal delivered from women with COVID-19.Result:s (1) Of the 16 pregnant women with COVID-19, 15 cases were ordinary type and 1 case was severe type. No one has progressed to critical pneumonia.The delivery method of the two groups was cesarean section, and the gestational age were (38.7±1.4) and (37.9±1.6) weeks,there was no significant difference between the two groups ( P>0.05). Also, there wee no significant differences in the intraoperative blood loss and birth weight of the newborn between the two groups (all P>0.05). (2) Ten cases of neonates delivered from pregnant women with COVID-19 were collected. The 2019-nCoV nucleic acid test were all negative.There were no significant differences in fetal distress, meconium-stained amniotic fluid, preterm birth, and neonatal asphyxia between the two groups (all P>0.05).(3) In the treatment of uterine contraction fatigue, carbetocin or carboprost tromethamine was used more in cesarean section for pregnant women with COVID-19 (1.3±0.6), compared with Non-COVID-19 group (0.5±0.7),the difference was statistically significant ( P=0.001). Conclusions:If there is an indication for obstetric surgery or critical illness of COVID-19 in pregnant women, timely termination of pregnancy will not increase the risk of premature birth and asphyxia of the newborn, but it is beneficial to the treatment and rehabilitation of maternal pneumonia. Preventive use of long-acting uterotonic agents could reduce the incidence of postpartum hemorrhage during surgery. 2019-nCoV infection has not been found in neonates delivered from pregnant women with COVID-19.

Objective:To investigate the regulatory effects of sphingosine kinase-2 (SphK2) on the function of activated astrocytes and the progression of experimental autoimmune encephalomyelitis (EAE) in mice.Methods:Primary mouse astrocytes were isolated from wild-type (WT) C57BL/6 mice and sphk2 gene knock-out ( sphk2 -/-) mice and stimulated in vitro with interleukin 17 (IL-17). Real-time PCR was used to measure the expression of inflammatory cytokines and chemokines at mRNA levels. Western blot and immunofluorescence were used to detect the expression of glial fibrillary acidic protein (GFAP) and the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). An EAE mouse model was constructed using myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) polypeptide. Western blot was used to detect the expression of GFAP and p-STAT3 at protein level and real-time PCR was used to detect the expression of inflammatory cytokines and chemokines at mRNA level in spinal cords. Hematoxylin-Eosin (HE) and Luxol fast blue (LFB) staining were used to observe the changes in inflammatory cell infiltration and demyelination in spinal cords. Results:Compared with the WT group, the phosphorylation of STAT3 was obviously reduced in in vitro activated mouse astrocytes of sphk2 -/- mice, and the expression of inflammatory cytokines and chemokines including monocyte chemotactic protein 1 (MCP-1), TNF-α and IL-6 at mRNA level was also significantly decreased. Compared with the WT EAE group, changes in the above-mentioned cytokines and relative proteins in sphk2 -/- EAE mice in vivo were similar to those in vitro. Moreover, inflammatory cell infiltration and demyelination were significantly reduced in spinal cords of sphk2 -/- EAE mice. However, no significant difference in in vitro or in vivo GFAP expression was observed between WT and sphk2 -/- mice. Conclusions:SphK2 might regulate the function of reactive astrocytes through STAT3 molecular pathway, thereby regulating the production of inflammatory cytokines and chemokines and participating in the pathological process of EAE.

Objective:The diagnostic values of urine free and fractionated catecholamine metabolites (including metanephrine MN and normetanephrine NMN, usually known as MNs) were established and their clinical value was evaluated.Methods:Using high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS), urine free MNs (f-MN, f-NMN) and fractionated MNs(t-MN, t-NMN) from 180 cases of non-pheochromocytoma and 54 cases of pheochromocytoma (PCC)patients were detected respectively.Receiver operating characteristic curve (ROC) was used to establish clinical reference cut-off value for different forms of MNs, and diagnostic efficacy of free, fractionated and total MNs was evaluated.Results:(1) The cut-off values of f-MN, f-NMN, t-MN and t-NMN were 47.8 μg/24 h, 52.3 μg/24 h, 224.9 μg/24 h and 664 μg/24 h, respectively. The cut-off values of total f-MNs and total t-MNs were 126 μg/24 h and 1 070 μg/24 h, respectively. (2) The correlation between f-MN and t-MN ( r=0.976, P<0.001), f-NMN and t-NMN ( r=0.940, P<0.001) was good. The area under ROC curve(AUC)of f-MN was lower than that of t-MN(0.579 vs 0.730, P<0.001), the sensitivity was slightly lower than that of t-MN((37.01% vs 51.85%, P=0.036), and the specificity was similar (99.44% vs 96.67%, P>0.05). There was no significant difference in sensitivity (90.74% vs 92.59%, P>0.05), specificity (99.44% vs 96.67%, P>0.05) and AUC (0.944 vs 0.959, P>0.05) between f-NMN and t-NMN. The combined diagnostic value of MN and NMN (total MNs) was higher than MN (free type:0.932>0.579, fractionation type: 0.960>0.730), which was similar to NMN. Conclusions:The diagnostic performance of urine free NMN or total MNs for PCC is similar to that of fractionated typewhich can meet the clinical needs.With few influencing factors, free type MNs may be used as an alternative indicator for PCC screening in the future.

Objective: To facilitate using the CRISPR/Cas9 gene editing system in human liver and gallbladder cancer cells, we established Cas9 stably expressed human liver and gallbladder cancer cell lines, and validated the gene editing activity of Cas9. Methods: Human liver cancer cell lines (Huh7, PLC/PRF/5, HepG2, Hep3b, SK-HEP-1 and Li-7), human cholangiocarcinoma cells (RBE) and human gallbladder cancer cells (GBC-SD) were infected with 3 Cas9-expressing lentivirus vectors (pLv-EF1α-Cas9-Flag-Neo, pLv-EF1α-Cas9-Flag-Puro, Cas9m1.1), respectively, and Cas9 stably expressed colonies were screened and selected. We extracted the genomic DNA and protein, validated the stable expression of Cas9 by using genomic polymerase chain reaction (PCR) and western blot. Three of cell lines were further infected with Lv-EF1α-mCherry. Then mCherry positive cells were sorted by flow cytometry and infected with designed guide RNA (gRNA) vectors which targeted mCherry gene. Subsequently the gene editing activity of Cas9 was detected by genomic PCR, fluorescence microscopic observation and flow cytometry analysis. Results: One hundred Cas9-expressing human liver and gallbladder cancer cell lines were selected. Among them, 35 cell lines expressed Cas9-Neo, 25 expressed Cas9-puro, and 40 expressed mutant Cas9 (mCas9). We also established 3 cell lines with stable expression of mCherry (Huh7-mCas9-M, PLC/PRF/5-Cas9-M and SK-HEP-1-Cas9-M). The results of genomic PCR and sequencing showed that by lentiviral infection with 2 types of designed gRNA, the long fragment deletion of mCherry gene was found in these 3 cell lines. Moreover, mCherry^-EGFP⁺ cells infected with 2 types of gRNA were observed by fluorescence microscope. The results of flow cytometry showed that mCherry^-EGFP⁺ cells accounted from 0.3% to 93.6%. Conclusion We successfully establish 100 human liver and gallbladder cancer cell lines with stable expression of Cas9 protein and validate their activities of gene editing.