ObjectiveTo study on the different therapeutic effects and potential mechanisms of Rhei Radix et Rhizoma（RH） before and after steaming with wine on constipation model mice. MethodsFifty-four male ICR mice were randomly divided into control group， model group， lactulose group（1.5 mg·kg^-1）， high， medium and low dose groups of RH and RH steaming with wine（PRH）（8， 4， 1 g·kg^-1）. Except for the control group， the constipation model was replicated by gavage of loperamide hydrochloride（6 mg·kg^-1） in the other groups. After 2 weeks of modeling， each administration group was gavaged with the corresponding dose of drug solution， and the control and model groups were given an equal volume of normal saline， 1 time/d for 2 consecutive weeks. After administration， the feces were collected for 16S rRNA sequencing， the levels of gastrin（GAS）， motilin（MTL）， interleukin-6（IL-6）， γ-interferon（IFN-γ） in the colonic tissue were detected by enzyme-linked immunosorbent assay（ELISA）， the histopathological changes of colon were observed by hematoxylin-eosin（HE） staining， flow cytometry was used to detect the proportion changes of CD4⁺， CD8⁺ and regulatory T cell（Treg） in peripheral blood. ResultsCompared with the control group， the model group showed significantly decrease in fecal number in 24 h， fecal quality and fecal water rate（P<0.01）， the colon was seen to have necrotic shedding of mucosal epithelium， localized intestinal glands in the lamina propria were degenerated， necrotic and atrophied， a few lymphocytes were seen to infiltrate in the necrotic area in a scattered manner， the contents of GAS and MTL， the proportions of CD4⁺， CD8⁺ and Treg were significantly reduced（P<0.01）， the contents of IL-6 and IFN-γ were significantly elevated（P<0.05， P<0.01）. Compared with the model group， the fecal number in 24 h， fecal quality and fecal water rate of high-dose groups of RH and PRH were significantly increased（P<0.05， P<0.01）， the pathological damage of the colon was alleviated to varying degrees， the contents of GAS， MTL， IL-6 and IFN-γ were significantly regressed（P<0.05， P<0.01）， and the proportions of CD4⁺ and CD8⁺ were significantly increased（P<0.01）， although the proportion of Treg showed an upward trend， there was no significant difference. In addition， the results of intestinal flora showed that the number of amplicon sequence variant（ASV） and Alpha diversity were decreased in the model group compared with the control group， and there was a significant difference in Beta diversity， with a decrease in the relative abundance of Lactobacillus and an increase in the relative abundances of Bacillus and Helicobacter. Compared with the model group， the ASV number and Alpha diversity were increased in the high-dose groups of RH and PRH， and there was a trend of regression of Beta diversity to the control group， the relative abundance of Lactobacillus increased， and the relative abundances of Bacillus and Helicobacter decreased. ConclusionRH and PRH can improve dysbacteriosis， promote immune system activation， inhibit the release of inflammatory factors for enhancing the gastrointestinal function， which may be one of the potential mechanisms of their therapeutic effect on constipation.

Objective To explore the mediating role of intolerance of uncertainty(IU)and moderating role of the negative emotion differentiation in the influence of perceived stress on anxiety among college students from a cognitive perspective.Methods A total of 271 participants were surveyed using the perceived stress scale,intolerance of uncertainty scale,depression anxiety and stress scale(Chinese version),and the test on negative emotional differentiation.SPSS 22.0 was used to perform descriptive statistics and correlation analyses and to test the moderated mediation model.Results Perceived stress affected anxiety and IU played a mediating role-perceived stress could affect anxiety through influencing IU.At the same time,the influence of IU on anxiety could be adjusted through the negative emotion differentiation.The higher the degree of negative emotion differentiation,the lower the degree of anxiety increase(β=0.17,t=5.70,P＜0.01).Conclusion It may be effective to develop training programs to reduce anxiety by regulating perceived stress,increasing acceptance of uncertainty,and improving the negative emotion differentiation,which can help individuals reduce anxiety by perceiving and adjusting anxiety-related emotional or cognitive factors in a timely manner.

Objective:To investigate the clinical effect of minimally invasive-locking block modified Krackow (MI-LBMK) and open giftbox technique in the treatment of Achilles tendon rupture.Methods:Fifty-six patients with Achilles tendon rupture from January 2016 to December 2018 were collected, including 54 males and 2 females, aged 40.7±9.4 years (range 26 to 65 years). The MI group (30 patients) used two minimally invasive incisions without exposing the rupture site, and the LBMK technique was used to repair the Achilles tendon. The open group (26 patients) used a posteromedial longitudinal incision and the giftbox technique was used to repair the rupture tendon. The Achilles tendon was repaired with 6-strand sutures in both groups. Early rehabilitation programs were adopted for postoperative rehabilitation, and regular follow-up (6 weeks, 3, 6, 12 and 24 months after operation) was performed to record the Achilles tendon resting angle (ATRA), American Orthopaedic Foot and Ankle Society ankle-hindfoot score (AOFAS), Achilles tendon total rupture score (ATRS). The rupture gap and cross-sectional area (CSA) were measured by MRI at 6 weeks and 3 months after surgery.Results:A total of 30 patients in the MI group and 26 in the open group were enrolled. The differences between the two groups in age, body mass index, interval from injury to operation, and tendon rupture site were not statistically significant ( P>0.05). All patients were followed up to 24 months after surgery. There were no wound complications in MI group, and 2 cases of superficial infection and 1 case of wound skin necrosis occurred in open group. There was no re-rupture in both groups. The relative ATRA of MI group was -6.32°±0.99°, -3.90°±1.05°, -2.38°±0.84°, -0.25°±1.37° at 3, 6, 12 and 24 months after operation, respectively. The relative ATRA of open group was -7.88°±3.71°, -6.16°±1.10°, -4.53°±0.95°, -3.01°±0.95° at 3, 6, 12 and 24 months after operation, respectively. The differences between the two groups were statistically significant ( P<0.05). The ATRS of minimally invasive group at 6 months and 12 months were 72.70±7.41 and 92.97±3.35 respectively, and the ATRS of open group at 6 months and 12 months were 68.08±6.64 and 90.85±4.27 respectively, and the differences were statistically significant ( P<0.05). The AOFAS of minimally invasive group at 6 months and 12 months were 88.60±2.76 and 93.83±1.98 respectively, and the AOFAS of open group at 6 months and 12 months were 85.77±3.20 and 92.08±2.64 respectively, and the differences were statistically significant ( P<0.05). The difference in the gap between the tendon rupture ends measured by MRI sagittal plane T2WI between the two groups was not statistically significant ( P>0.05). The cross-sectional area of Achilles tendon in the MI group was higher than that of the open group at 12 weeks ( P<0.05). Conclusion:The MI-LBMK technique may protect the peritendon tissue and has fewer complications, and can enable the patient to return to daily life faster, with lower postoperative Achilles tendon elongation and better recovery of Achilles tendon function.

Objective:To compare the efficacy of 3D-printed guide plate assisted versus freehand placement of cannulated screws for the treatment of Sanders type II and III calcaneal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 29 patients with Sanders type II and III calcaneal fractures admitted to Chonggang General Hospital from June 2020 to October 2022. Among them, there were 18 males and 11 females, with an age range of 22-69 years [(40.1±11.5)years]. Nineteen patients were treated with individualized 3D-printed guide plate assisted placement of cannulated screws (3D-printed group) and 10 were treated with freehand placement of cannulated screws (freehand group). The surgical time, fluoroscopy times, postoperative 6-month calcaneal morphology (length, width, height, B?hler angle and Gissane angle), and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Maryland functional score assessed at 3, 6 months after operation and at the final follow-up were compared between the two groups. The incision healing and complications were observed.Results:The patients were followed up for 6-24 months [(11.3±2.5)months]. The surgical time and fluoroscopy times in the 3D-printed group were (53.4±9.1)minutes and (7.3±1.1)times, respectively, which were shorter than (90.2±16.0)minutes and (16.0±3.2)times in the freehand group (all P<0.01). At 6 months after operation, there was no significant difference in calcaneal length between the two groups ( P>0.05); the calcaneal width, height, B?hler angle and Gissane angle in the 3D-printed group [(34.0±1.8)mm, (47.2±1.6)mm, (27.8±1.0)°, (129.2±2.8)°] were superior than those in the freehand group [(37.5±2.0)mm, (43.0±2.7)mm, (25.8±1.5)°, (125.9±2.5)°] (all P<0.01). At 3, 6 months after operation and at the final follow-up, the values of AOFAS ankle-hindfoot score in the 3D-printed group [(72.2±2.3)points, (79.7±2.3)points, (86.5±4.4)points] were higher than those in the freehand group [(64.2±6.9)points, (73.4±4.2)points, (81.8±3.1)points] (all P<0.05); the values of Maryland score in the 3D-printed group [(71.4±7.7)points, (84.7±2.6)points, (91.5±2.5)points] were higher than those in the freehand group [(65.2±5.6)points, (79.1±3.8)points, (87.1±2.9)points] (all P<0.05). All surgical incisions were healed in stage I. In the 3D-printed group, there were no complications regarding infection, iatrogenic vascular or nerve injury, or fixation failure after surgery. In the freehand group, one patient with lateral sural cutaneous nerve injury was resolved spontaneously without specific treatment. Conclusion:Compared with freehand placement of cannulated screws, 3D-printed guide plate assisted placement of percutaneous placement has the advantages of shorter surgical time, fewer fluoroscopy times, lower reduction loss, better ankle joint function recovery, and less complications in treating Sanders type II and III calcaneal fractures.

Objective:To introduce a new TIANJI robot assisted targeted puncture technique, and discuss the feasibility and clinical effect of transforaminal percutaneous endoscopic lumbar discectomy (tPELD) using this technique.Methods:The first 14 consecutive cases of single level lumbar disc herniation who underwent robot assisted tPELD procedure were retrospectively analyzed. The mean age was 46.3±16.0 years old (ranged from 16-72). After data transferred from C-arm to robot system and automatic registration, surgeons made plans of the trajectory on robot system based on intraoperative 3-dimensional images of lumbar spine. Move robotic arm to planned position, guide an accurate puncture pathway and establish working cannula. 25 consecutive patients who underwent conventional C-arm assisted tPELD surgery during the same period of time were assessed as the controlled group. The mean age was 45.5±13.7 years old (ranged from 16-68). All patients were followed up for 12 months. Clinical effect was assessed by visual analogue scale (VAS), Oswestry disability index (ODI) and Modified Macnab criteria. Intraoperative parameters and surgery-related complications were recorded.Results:The baseline data of age, surgical level, types of herniation, preoperative VAS scores and ODI had no significant difference between two groups ( P>0.05). In robot group, one case was converted to open microdiscectomy during operation due to technical failure. The other thirteen cases had successful robot assisted tPELD surgeries and were assessed accordingly. The new technique had good clinical outcomes. The immediate post-operative VAS score 2.85±1.79 and the last follow-up VAS score 1.50±1.04 were both significantly decreased than that before surgery 7.62±0.92 ( F=69.747, P<0.01); the last follow-up ODI 18.89%±12.16% was significantly reduced from the pre-operative ODI 71.19%±12.12% ( t=15.430, P<0.01). Between two groups, the immediate post-operative VAS score ( t=0.568, P=0.574), the last follow-up VAS score ( t=0.713, P=0.481), and last follow-up ODI had no significant difference ( t=0.171, P=0.865). The excellent or good rate of modified Macnab criteria at the last follow-up was 92.30% in robot group, comparing to 84.0% in controlled group. The fluoroscopic times during surgery of robot group 8.8±5.5 was significantly lowered the in controlled group 21.3±8.3 ( P<0.01). One case in robot group and two cases in controlled group had recurrence during follow-up period (recurrence rate 7.7% vs. 8.3%). However, there was no significant complications such as nerve root injury, dura injury or increased intracranial pressure in both groups. Conclusion:This study confirmed the feasibility of this new technique. Preliminary results indicated that TIANJI robot could help to build an easy, accurate and safe procedure of tPELD surgery.

To develop Senecavirus A (SVA) virus-like particles (VLPs), a recombinant prokaryotic expression plasmid pET28a-SVA-VP031 was constructed to co-express SVA structural proteins VP0, VP3 and VP1, according to the genomic sequence of the field isolate CH-FJ-2017 after the recombinant proteins were expressed in E .coli system, and purified by Ni+ ion chromatographic method. The SVA VLPs self-assemble with a high yield in vitro buffer. A typical VLPs with an average diameter of 25-30 nm which is similar to native virions by using TEM detection. Animals immunized by SVA VLPs shown that the VLPs induced high titers neutralizing antibodies in Guinea pigs. This study indicated that the VLPs produced with co-expressing SVA structural proteins VP0, VP3 and VP1 in prokaryotic system is a promising candidate and laid an important foundation for the development of a novel SVA VLPs vaccine.
Animals ; Antibodies, Neutralizing ; Escherichia coli/genetics* ; Genomics ; Guinea Pigs ; Picornaviridae/genetics*

Objective:To explore an ideal method for establishing a mouse model of chronic atrophic gastritis (CAG).Methods:CAG mouse models were established with five different modeling methods ( N-methyl- N′-nitro- N-nitrosoguanide (MNNG), sodium salicylate, sodium deoxycholate, Helicobacter pylori infection, and combinations of them) in BALB/c and C57 mice. The effect of each modeling method was evaluated by histological observation of gastric mucosa, plasma biochemical parameters, inflammatory response score, and the expression of anti-inflammatory factors. Results:The results of histological observation of gastric mucosa showed that all of the 5 methods could successfully establish CAG mouse models. In BALB/c mice, compared with the healthy control group, significant features of CAG accompanied with intestinal metaplasia was found in the model group established by combination of MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate. From the results of serological detection, compared with the normal control group, the mRNA expression levels of related anti-inflammatory factors interleukin-2, interleukin-10, interleukin-13 and growth differentiation factor-15 of each model group decreased, which indicated that the mice of each CAG model group had different degrees of inflammation. The results of plasma biochemical parameters indicated that plasma gastrin of each group decreased and the ratio of pepsinogen Ⅰ and pepsinogen Ⅱ significantly dropped. The above results demonstrated that in BLAB/c mice, MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate was better than other four modeling methods. For C57 mice, it was also found that simple chemical drug mutagenesis and Helicobacter pylori replication method both could successfully establish CAG models. No matter from pathological observation, relative expression of anti-inflammatory factors and analysis of plasma biochemical parameters, the effects of combination of the two methods was better. Conclusion:The CAG mouse model established by MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate can provide a certain reference for the establishment and application of mouse model in CAG experiments in the future for pharmacological research.

Objective:To evaluate the feasibility of surgery for upper cervical spine anomaly instability achieved by readjusting the clivus-axial angle (CAA) and evaluating available screw trajectories based on preoperative virtual simulation reduction using 3D CT reconstruction.Methods:From January 2014 to September 2019, 52 patients (28 males, 24 females; mean age 46.7 years; range 18-64 years) with upper cervical spine anomalies were enrolled in a retrospective study. DICOM data of preoperative CT scanning of cervical spine were imported into Mimics 15.0 to reconstruct the 3D model of atlantoaxial joint. The target range of CAA was set with the value measured in fusing images of extension view of cervical spine as the upper bound; and 140°, the lower limit of normal CAA range, was set as the lower bound. The trajectories of screws were sought within the target range of CAA. The exact CAA value of the available trajectories was set as the target angle of intraoperative reduction . The preoperative, postoperative and simulated target CAA values were compared. The accuracy of screw placement was evaluated according to the Gertzbein-Robbins scale. The pre- and postoperative neck disability index (NDI) and Nurick scale were also compared.Results:Among 52 patients, available screw trajectories existed in 35 patients. With the validation under computer-assisted navigation as the gold standard, 37 patients underwent Magerl screws fixation (Magerl group) while the other 15 patients underwent screw-rods fixation (screw-rods group). In Magerl group, no significant difference was found between simulated target CAA values (150.1°±6.6°) and postoperative CAA values (149.0°±6.5°)( t=1.194, P=0.240); postoperative CAA values were significantly larger than preoperative CAA values (124.0°±8.9°)( t=-13.499, P< 0.001). In screw-rods group, no significant difference was found between simulated target CAA values (150.4°±5.6°) and postoperative CAA values (150.2°±6.1°) ( t=0.319, P=0.754); postoperative CAA values were also significantly larger than preoperative CAA values (121.9°±8.3°)( t=-12.431, P< 0.001). Follow-up data were obtained in all 52 patients, with a mean follow-up time of 1.7±0.9 years (range, 0.5-4 years). A total of 106 screws were placed into 52 patients, including 74 in Magerl group and 32 in screw-rods group. According to the Gertzbein-Robbins scale, 88 and 15 were classified into grade A and B respectively; and 97.1% (103/106) of screws were acceptable. No severe complications such as vertebral artery injury and low cranial nerve palsy occurred. The postoperative NDI (9.7±6.4 vs. 27.4±8.7) and Nurick scale (1.1±1.4 vs. 2.6±1.3) decreased significantly compared to preoperative ones, respectively. Conclusion:Preoperative virtual simulation can evaluate available screw trajectories and provide the target CAA values for guidance of intraoperative reduction. Based on this, readjusting the CAA under computer-assisted navigation were safe and feasible, and warranted the possibility of a single posterior reduction and fusion approach for reducible upper cervical anomaly instability.

Spinal surgery is a technically demanding and challenging procedure because of the complicated anatomical structures of the spine and its proximity to several important tissues. Surgical landmarks and fluoroscopy have been used for pedicle screw insertion but are found to produce inaccuracies in placement. Improving the safety and accuracy of spinal surgery has increasingly become a clinical concern. Computerassisted navigation is an extension and application of precision medicine in orthopaedic surgery and has significantly improved the accuracy of spinal surgery. However, no clinical guidelines have been published for this relatively new and fast-growing technique, thus potentially limiting its adoption. In accordance with the consensus of consultant specialists, literature reviews, and our local experience, these guidelines include the basic concepts of the navigation system, workflow of navigation-assisted spinal surgery, some common pitfalls, and recommended solutions. This work helps to standardize navigation-assisted spinal surgery, improve its clinical efficiency and precision, and shorten the clinical learning curve.

Objective:To describe the characteristics of genotype and phenotype in 3 families with X-linked retinoschisis (XLRS) due to RS1 mutations. Methods:A cross-sectional approach was adopted.Three XLRS families at the Ningxia Eye Hospital from October 2017 to March 2019 were included.Clinical data and peripheral blood of patients and related families were collected and clinically staged were formulated through a comprehensive eye examination.The disease-causing genes screened by panel sequencing underwent conservative analysis, pathogenicity analysis and protein structure prediction by software tools.Analysis of the mutations pathogenicity was performed according to the American College of Medical Genetics and Genomics guidelines.The research was approved by Medical Ethics Committee of the People's Hospital of Ningxia Hui Autonomous Region and followed the Declaration of Helsinki.Written informed consent was obtained from each participant.Results:Total 5 young male patients and 1 middle-aged patient in these three families.The optical coherence tomography(OCT) findings of 5 young patients showed typical macular retinoschisis, which were characterized by stage I of XLRS.One middle-aged patient (Ⅱ-9) showed a stage Ⅲ lesion of macular atrophy.The mutations of c. 668G>A, c.618G>A and exon 1 deletion in RS1 gene were found in the three families.C223 and W206 were verified to be highly conserved in mammals and were predicted to be pathogenic mutations by software and the change of protein structure.Conservation analysis and prediction of protein structure were not performed for the mutation of exon 1 deletion.All the mutations were pathogenic variants according to the ACGM guidelines. Conclusions:Mutations of c. 668G>A/p.C223Y, c.618G>A/p.W206X and exon 1 deletion in RS1 gene are pathogenic mutations in Chinese XLRS families.The combination of Panel sequencing with pathogenicity analysis and protein structure prediction have important effect to diagnosis and identify the causative genes for the hereditary retinal diseases.