ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

There are millions of patients with taxane-induced peripheral neuropathy （TIPN）， and there is no effective treatment or prevention measure in clinical practice. The occurrence of TIPN may be related to the dosage form of paclitaxel drugs， genetic and molecular markers， drug dosage and chemotherapy cycle， patient factors， etc. At present， drugs for treating TIPN mainly include those that inhibit axonal degeneration （such as dosazosin， tamsulosin）， prevent mitochondrial dysfunction （such as glutathione trisulfides， antioxidants α -lipoic acid）， improve calcium imbalance in the internal environment （Shaoyao gancao decoction， N-type voltage-gated calcium channel inhibitor IPPQ）， and inhibit neuroinflammation （such as chemokine inhibitors and selective interleukin-8 receptor inhibitors DF2726A）. Further exploration of drug treatment strategies targeting different induction mechanisms is expected to become a new direction for precise clinical prevention and personalized treatment of TIPN.

Humans ; SARS-CoV-2 ; COVID-19 ; Antibodies ; Antibodies, Viral/therapeutic use* ; Antibodies, Neutralizing/therapeutic use*

Objective:To explore the effect and mechanism of interleukin-35 gene modified mesenchyma stem cells(MSC)on ameliorating cardiac allograft rejection and prolonging graft survival of transplanted heart in mice.Methods:In this study, IL-35-MSC secreting IL-35 continuously and steadily were successfully constructed in vitro. Abdominal heterotopic heart transplantation model was established successfully. And they were randomly divided into syngeneic control group; saline control group, MSC treatment group and IL-35-MSC experimental group(n=12 each). Six mice were randomly selected for sacrificing at Day 5 post-operation for detecting the related indicators in each group: Hematoxylin eosin staining was used for pathological examination. Enzyme-linked immunosorbent assay(ELISA)was employed for detecting the concentration of IL-35 in peripheral blood and the proportion of T lymphocyte subsets in spleen was analyzed by flow cytometry(FCM). Then the remaining mice were used for recording the graft survival.Results:The model of abdominal heterotopic heart transplantation in mice was successfully constructed. As compared with saline control group(6.50±0.55 d)and MSC treatment group(12.00±0.89 days), IL-35-MSC significantly alleviated rejection after transplantation and effectively prolonged the survival time of graft(18.50±1.64 days)(n=6, P<0.01). As compared with other groups, percentage of Th17 cells and Th1/Th2 ratio in spleen decreased significantly while the proportion of CD4 + Foxp3 + Treg increased significantly in IL-35-MSC experimental group at Day 5 post-transplantation(n=6, P<0.01). Conclusions:IL-35-MSC may alleviate cardiac allograft rejection and prolong graft survival. And cellular immunotherapy based upon IL-35-MSC may provide a new approach for inducing immune tolerance.

BACKGROUND:Previous study has found that hsa-miR-182 is probably related to the apoptosis-related genes such as cytochrome C (Cycs C) and calcineurin subunit CnB (PPP3R1) in nucleus pulposus cells.
OBJECTIVE:To determine whether miR-182 plays a regulatory role in nucleus pulposus cel apoptosis by detecting the relative gene expression levels after transfecting miR-182 with Cycs C and PPP3R1 into nucleus pulposus cel s via plasmid delivery.
METHODS:After a bioinformatics prediction about miR-182, miR-182 and target genes were transfected into the nucleus pulposus cel s, and at the same time, blank control group was established. Then the expression levels of the target genes were detected through cel lysis.
RESULTS AND CONCLUSION:miR-182 significantly inhibited the expression of Cycs C in nucleus pulposus cel s compared with the blank control group (P<0.05). Compared with the blank control group, miR-182 made no inhibitory effect on the expression of PPP3R1. These findings suggest that miR-182 may play a regulatory part in nucleus pulposus cel apoptosis by inhibiting the expression of Cycs C.

BACKGROUND:We have designed and manufactured a novel artificial cervical vertebra and intervertebral complex (ACVC) which combines the cervical titanium cage with the artificial cervical disc, and also developed the ACVC with a hydroxyapatite biocoating (ACVC-HA). OBJECTIVE:To evaluate biomechanical properties of the joint system, and the role of HA coating in promoting osseointegration and long-term stability. METHODS:Twenty-four goats were randomly divided into three groups and underwent the anterior C2/3 and C3/4 discectomy, and C3 subtotal corpectomy, fol owed by ACVC implantation (group 1) and ACVC-HA implantation (group 2), and given no intervention (black control group), respectively. group. At 12 weeks after surgery, C1-5 samples were col ected to undergo biomechanical tests and histological staining. RESULTS AND CONCLUSION:Prior to the fatigue test, compared with the blank control group, the range of motion and neural zone of groups 1 and 2 in the directions of flexion-extension and lateral bending showed no significant differences, but the above indicators were significantly increased in the direction of rotation (P<0.05). Additional y, the stiffness in al three directions was significantly lower than that in the blank control group (P<0.05). There were no significant differences in the range of motion and neural zone in al directions between groups 1 and 2. Similar results were found after the fatigue test. The histological staining showed that both two implants had good biocompatibility and abradability, but more new bone formed on the ACVC-HA. These results suggest that ACVC can effectively reconstruct the motor function of the cervical spine after decompression. Furthermore, HA coating can markedly improve bone-implant interface to promote osseointegration.

Objective Todetecttheexpressionlevelsoftumornecrosisfactor-α(TNF-α)and interleukin-6(IL-6)inintracranialaneurysms.Methods Sixteenconsecutivepatients(aneurysm group)with intracranial aneurysm confirmed by digital subtraction angiography (DSA)and clipped by microneurosurgery were enrolled retrospectively. A total of 19 trauma patients without vascular disease confirmed by CT and magnetic resonance imaging (MRI)in the same period were used as a control group. Hematoxylin-eosin (HE)staining and immunohistochemical staining were used to detect the aneurysm wall tissue and the colored portions of TNF-α and IL-6 in normal vessel wall,the mean value of optical density after its expression was analyzed,and the intensity of staining was compared. Results (1)Each layer of artery walls of the control group had no obvious TNF-α and IL-6 expression. The inner,media and out membranes of the aneurysm wall tissue of the aneurysm group had positive expression of TNF-αand IL-6. (2)The mean optical densities of TNF-α and IL-6 in patients of the aneurysm group were 0. 182 ± 0. 069 and 0. 148 ± 0.062 respectively,and they were higher than 0. 144 ± 0. 031 and 0. 105 ± 0. 020 of the control group. The differences were statistically significant (all P<0. 05). (3)The mean optical densities of TNF-α expression of each layer of the inner,media and out membranes in the aneurysm walls were 0. 224 ± 0. 071,0. 134 ± 0. 040,and 0. 106 ± 0. 065,respectively. There were significant differences (P<0.01). (4)The mean optical density expressed by IL-6 in the out membrane of the aneurysm walls was lower than the media and inner membranes (0. 096 ± 0. 018 vs. 0. 145 ± 0. 050,and 0. 148 ± 0. 070). There were significant differences (P<0. 05). (5)The results of Spearman correlation analysis showed that the mean optical density of TNF-αof the aneurysm group was positively correlated with that of IL-6 (r=0. 452, P<0.05).Conclusion TheexpressionlevelsofTNF-αandIL-6intheaneurysmwalltissueare higher,and they may be involved in intracranial aneurysm formation and rupture.

OBJECTIVETo explore the value of preoperative evaluation with three-dimensional endoanal ultrasonography (3D-EAUS) for anal fistula in order to provide preoperative assessment for anal fistula.

METHODSOne hundred patients diagnosed with anal fistula undergoing surgery between March 2012 and March 2013 in our department were prospectively enrolled. All the patients were randomly divided into the ultrasound group and the control group with fifty patients in each group. The ultrasound group received 3D-EAUS and the control group received routine examinations (digital examination and probe) to assess the position of the internal opening, the type of fistula and secondary tracks, respectively. The concordance rate of the preoperative assessment and intraoperative exploration was evaluated between the two groups.

RESULTSThe accuracy of identifying internal opening was 96.0% for the ultrasound group and 82.0% for the control group with statistically significant difference (P=0.02). The accuracy of identifying internal opening for simple anal fistula was similar (95.0% vs. 91.3%, P=1). For complex anal fistula, the accuracy was also higher in the ultrasound group (96.7% vs. 74.1%, P=0.025). The accuracy of fistula classification was 78.0% for the ultrasound group and 96.0% for the control group with significant difference (P=0.01). The accuracy of identifying a second track was higher in the ultrasound group (96.0% vs. 82.0%, P=0.025).

CONCLUSIONSIt is significantly superior for 3D-EAUS to detect the internal opening, fistula classification and identification of a second track in complex anal fistulas as compared to conventional examination. 3D-EAUS should be recommended as a preoperative assessment for anal fistula, especially for complex one.

Objective To explore the value of preoperative evaluation with three-dimensional endoanal ultrasonography (3D-EAUS) for anal fistula in order to provide preoperative assessment for anal fistula. Methods One hundred patients diagnosed with anal fistula undergoing surgery between March 2012 and March 2013 in our department were prospectively enrolled. All the patients were randomly divided into the ultrasound group and the control group with fifty patients in each group. The ultrasound group received 3D-EAUS and the control group received routine examinations (digital examination and probe) to assess the position of the internal opening, the type of fistula and secondary tracks, respectively. The concordance rate of the preoperative assessment and intraoperative exploration was evaluated between the two groups. Results The accuracy of identifying internal opening was 96.0%for the ultrasound group and 82.0% for the control group with statistically significant difference (P=0.02). The accuracy of identifying internal opening for simple anal fistula was similar (95.0% vs. 91.3%, P=1). For complex anal fistula, the accuracy was also higher in the ultrasound group (96.7%vs. 74.1%, P=0.025). The accuracy of fistula classification was 78.0% for the ultrasound group and 96.0% for the control group with significant difference (P=0.01). The accuracy of identifying a second track was higher in the ultrasound group (96.0% vs. 82.0%, P=0.025). Conclusions It is significantly superior for 3D-EAUS to detect the internal opening , fistula classification and identification of a second track in complex anal fistulas as compared to conventional examination. 3D-EAUS should be recommended as a preoperative assessment for anal fistula , especially for complex one.