BACKGROUND:Previous studies have confirmed that Wen-Shen-Tong-Du Decoction can promote the recovery of spinal cord injury by inhibiting pyroptosis of splenic B cells,promoting the phagocytosis of myelin debris by microvascular endothelial cells,affecting the migration and infiltration of microglia,promoting the recovery of damaged neurons,and decreasing neuronal apoptosis after spinal cord injury,but the mechanism of this is still not clear. OBJECTIVE:To investigate the effect of Wen-Shen-Tong-Du Decoction on the triggering receptor expressed on myeloid cells 2(TREM2)and PI3K/Akt signaling pathways in mice following spinal cord injury. METHODS:Thirty-six C57BL/6 mice were selected and randomly divided into a sham-operation group,a model group and a Wen-Shen-Tong-Du Decoction group,with 12 mice in each group.In the model and Wen-Shen-Tong-Du Decoction groups,mouse models of T10 spinal cord injury were prepared by the modified Allen's method.On the 1st day after modeling,the Wen-Shen-Tong-Du Decoction group was given Wen-Shen-Tong-Du Decoction by gavage,and the sham-operation group and the model group were given saline by gavage once a day for 28 days.During the drug administration period,mouse motor function was evaluated by Basso Mouse Scale score and inclined plane test.On the 7th and 28th days after modeling,hematoxylin-eosin staining was used to observe the histopathological changes in the spinal cord tissue of the mice;immunofluorescence double staining was used to detect the protein expression of ionized calcium binding adaptor molecule 1(IBA1)and TREM2;and western blot assay was used to detect the expression of TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,Bax and Caspase3 in spinal cord tissue. RESULTS AND CONCLUSION:Basso Mouse Scale scores and inclined plane test results indicated that the motor function of the mouse hindlimbs was declined after spinal cord injury,and Wen-Shen-Tong-Du Decoction significantly improved motor function in mice with spinal cord injury.Hematoxylin-eosin staining results revealed that Wen-Shen-Tong-Du Decoction significantly ameliorated the pathological structure of spinal cord tissue compared with the model group,manifesting as reduced degrees of dorsal white matter and neuronal atrophy,decreased cytoplasmic vacuolization,and reduced inflammatory cell infiltration.Immunofluorescence double staining results showed that on the 7th day after modeling,the protein expression of IBA1 and TREM2 in the model group was lower than that in the sham-operation group(P＜0.05),and the protein expression of IBA1 and TREM2 in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P＜0.05);on the 28th day after modeling,the protein expression of TREM2 in the model group was lower than that in the sham-operation group(P＜0.05),and the protein expression of TREM2 in the spinal cord tissue of the mice in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P＜0.05).Western blot results analysis demonstrated that on the 7th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,and Bcl2/Bax(P＜0.05),as well as a significant increase in p-Akt,Bax and p-Akt/Aktp-PI3K(P＜0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,p-PI3K/PI3K,p-Akt/Ak,and Bcl2/Bax(P＜0.05),as well as a significant decrease in Bax and Caspase3 protein expression(P＜0.05).On the 28th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2 and Bcl2/Bax(P＜0.05),as well as a significant increase in Bax protein expression(P＜0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,Akt,p-Akt,Bcl2,and Bcl2/Bax(P＜0.05),as well as a significant decrease in Bax protein expression(P＜0.05).To conclude,Wen-Shen-Tong-Du Decoction may activate the PI3K/Akt signaling pathway by up-regulating the expression of TREM2 protein in microglia,and then inhibit neuronal apoptosis,thus exerting neuroprotective effects and promoting the repair of spinal cord injury.

ObjectiveTo explore the effects and possible mechanism of Wenshen Tongdu Formula （温肾通督方， WTF） on spinal cord injury. MethodsThirty-six C57BL/6 female mice were randomly divided into sham operation group， model group and WTF group， with 12 mice in each group. The spinal cord injury model was established in the model group and the WTF group using the modified Allen's method， while in the sham operation group the spinal cord was only exposed. Since the 1st day after surgery， 50 g/（kg·d） of WTF solution was given to the WTF group by gavage， while 20 ml/（kg·d） of normal saline was given to the sham operation and model group by gavage， all for 14 days. Before surgery and on the 1st， 7th， and 14th days after surgery， the motor function of the mice was evaluated using the inclined plane test and hind limb motor function score （by BMS）. On the 3rd day after surgery， the nerve electrophy-siology was detected through electromyography and motor evoked potential； the spleen length was measured， and B cells in the spleen were sorted by magnetic beads； the differential expression of proteins were detected through proteomics technology； and the protein expression of mitochondrial outer membrane transport porin 20 （Tom20） and downstream cleaved caspase-3 in spleen B cells were measured using Western blotting. On the 14th day after surgery， MRI was used to observe the recovery of the spinal cord. ResultsCompared to those in the sham operation group at the same time， the BMS scores and subscores and the inclined plane test angle in the model group were reduced on the 1st， 7th and 14th days after surgery； the peak value of electromyogram and motor evoked potential were reduced， and the spleen length was shortened， while the expression of Tom20 and cleaved caspase-3 increased in splenic B cells increased （P＜0.05）. Compared to those in the model group at the same time， the BMS subscores on the 14th day and the angle of the inclined plane test on the 7th and 14th days after surgery increased in the WTF group； the peak value of electromyography and motor evoked potential， as well as the length of spleen increased， and the expression of Tom20 and cleaved caspase-3 decreased （P＜0.05）. The proteomics results showed that there were 100 differential proteins in the WTF group versus the model group， of which 37 were up-regulated and 63 were down-regulated. GO enrichment analysis showed that differential proteins mainly played their roles in oxygen binding， exogenous apoptosis negative feedback， zinc ion response， and oxygen transport. KEGG enrichment analysis showed that differential proteins were mainly concentrated in metabolic pathways， Huntington's disease， oxidative phosphorylation and other pathways. Subcellular localization showed that differential proteins were associated with mitochondria. Magnetic resonance imaging on the 14th day after surgery showed that the spinal cord structure of the mice in the sham operation group was intact， and the segments were clear， with normal spinal cord signal； the low signal area in the spinal cord injury area increased in the model group， and the spinal cord became significantly thinner； the injured segment had obvious depression in the WTF group， but the structure was more complete than that in the model group. ConclusionWTF may promote spinal cord injury repair by regulating immune function， and its mechanism may be related to inhibiting pyroptosis of spleen B cells.

Objective To investigate the anti-inflammatory effect and mechanism of sappanone A(SA)on lipopolysaccharide(LPS)-induced RAW264.7 cell model based on JAK2-STAT3 signaling pathway.Methods MTT assay was used to detect the effects of sappanone A,LPS and AG490 on RAW264.7 cell viability.The LPS-induced inflammatory model in RAW264.7 cells was established,and the secretion level of interleukin-6(IL-6)in the supernatant was detected by ELISA.mRNA expressions of IL-6,Janus kinase 2(JAK2)and signal transducer and activator of transcription 3(STAT3)were tested by RT-PCR.The protein expressions of JAK2,phosphorylated JAK2(p-JAK2),STAT3 and phosphorylated STAT3(p-STAT3)were determined by Western Blot.Results Compared with control group,IL-6 secretion level was significantly increased,mRNA expressions of IL-6,JAK2 and STAT3 were up-regulated,and protein expressions of p-JAK2 and p-STAT3 were increased(all P＜0.01)in model group.Compared with model group,high-dosed sappanone A(5 μg·mL-1)was significantly decreased the secretion of IL-6,down-regulated the mRNA expressions of IL-6,JAK2 and STAT3,and inhibited the protein expressions of p-JAK2 and p-STAT3(all P＜0.01).Conclusion Sappanone A may play an anti-inflammatory role by inhibiting the JAK2-STAT3 signaling pathway and hence inhibiting the secretion of IL-6.

As a kind of mechanical effector cells, chondrocytes can produce a variety of physical and chemical signals under the stimulation of multiaxial load
Apoptosis ; Cells, Cultured ; Chondrocytes ; Stress, Mechanical

Objective: To setup a custom-made gait simulator, and to provide an efficient tool for biomechanics research of ankle and foot. Methods: From November 2017 to April 2018, a total of 6 fresh frozen specimens of the foot and ankle were collected. The donated specimens, free of diseases in the foot and ankle part, were from the Department of Anatomy, Shanghai Medical College of Fudan University. Donors were 3 males and 3 females, aged from 48 to 69 years old, with an average age of 58.8 years old. The nine tendons in the foot were divided into 4 bundles, including anterior group comprised of tibialis anterior (TA), extensor hallucis longus (EHL) and extensor digitorum longus (EDL). Posterior group comprised of Achilles's tendon (AT). Medial group comprised of tibialis posterior (TP), flexor hallucis longus (FHL), and flexor digitorum longus (FDL). Lateral group comprised of peroneus brevis (PB) and peroneus longus (PL). A custom-made gait simulator was set up by using four independent electro motors to actuate 4 bands of tendons in the foot and another six motors to control tibia to achieve 6 degree-freedom parallel mechanism. And a hydraulic machine was used to provide axial pressure along tibia. Gait cycle of six fresh frozen cadaver feet was reproduced using this machine, and the kinematics data of ankle movement and ground reaction force (GRF) data was collected. By comparing the data above with the normal human gait data, the simulation results were analyzed to explore the clinical usage of this machine. Results: On the sagittal plane, the ankle appeared to plantarflexion at the beginning of gait, and then turned to dorsiflexion after the max plantar flexion (about 10°) at 18% of gait cycle. At the 40% gait cycle, ankle joint was in neutral position and reached its max dorsiflexion (about 22°) at 83% gait cycle. On the coronal plane, ankle joint appeared inversion at the beginning and eversion afterwards with 10° range of change. On the horizontal plane, movement of ankle joint was small. Results showed that the first peak of vertical ground reaction force can reach to 1.1-1.3 times of bodyweight at 25% of gait cycle and the second peak appeared little lower at 70% of gait cycle. The GRF in posterior direction reached its peak at 30%, and then turned to anterior with its peak at 83% gait cycle. The GRF had small variation at the internal to external direction.All of the coefficients of multiple correlations (CMC) of GRF and ankle joint movements were close to or greater than 0.90. Conclusion This custom-made gait simulator has good gait simulation ability with high intra repeatability in respect of ankle rotation and ground reaction force, and can satisfy the request for ankle and foot biomechanics research.

OBJECTIVE: To summarize the clinical characteristics and treatment of type Ⅲ-b congenital intestinal atresia (CIA). METHODS: The clinical data of 12 type Ⅲ-b CIA treated in the Children's Hospital of Zhejiang University School of Medicine from January 2015 to December 2017 were analyzed retrospectively. RESULTS: Of the 12 patients diagnosed as type Ⅲ-b CIA in operation, treatment was refused during operation by their parents in 2 cases. For one child, only the proximal intestine was partly resected in the first operation, dilatation and dysplasia of the duodenum was diagnosed and total duodenum was resected and sutured in the second operation, as the child had postoperative intestinal obstruction. For one child, due to the long distal normal intestine, distal apple-peel like intestine was partly resected without mesenteric reformation. For the rest 8 children total duodenum resection and mesenteric reformation were performed. During the postoperative follow-up, one case was early rejected for further treatment by parents, one case died from complex congenital heart disease, 5 cases had the complication of short bowel syndrome. All 8 survival children received parenteral nutrition support after operation, 5 of whom received parenteral nutrition support for more than 42 days, and they were followed up for 1-3 years after discharge. The short-time efficacy was satisfactory. CONCLUSIONS For children with type Ⅲ-b CIA, the distal apple-peel like intestine should be preserved as much as possible, the mesenteric reformation should be performed and the proximal dilated bowel should be partly resected and sutured. Postoperative nutritional support and early intestinal rehabilitation contribute to the compensation for rest intestines.
Child ; Humans ; Intestinal Atresia ; complications ; surgery ; therapy ; Intestines ; surgery ; Parenteral Nutrition ; Retrospective Studies ; Short Bowel Syndrome ; complications ; Treatment Outcome

Objective: To explore the mid-term efficacy of derotation of talus and medial column fusion for stage II-V Müller-Weiss disease (MWD). Methods: Data of thirty-six patients (38 feet) with MWD treated by derotation of talus and talona-vicular (TN) or talonavicular-cuneiform (TNC) arthrodesis in our center during January 2008 to December 2016 were retrospective-ly analyzed. There were 3 males (3 feet) and 33 females (35 feet) with an average age of 54.0±9.2 years old (range, 32-80 years old); there were 11 right feet and 27 left feet. According to Maceira and Rochera staging system, there were 9 stage II cases, 11 stage III cases, 10 stage IV cases and 8 stage V cases. The external rotation of talus was intensively focused. After the TN articular surface was debrided, an "H" shape plate and a hollow lag screw were used to firmly fix the TN or TNC joint. American Orthopae-dic Foot and Ankle Society Scale (AOFAS), visual analog scale (VAS) and relative radiological parameters were evaluated preoper-atively and during follow-up. Results: The mean follow-up duration was 37.2 months (range, 25-113 months). The latest follow-up showed satisfactory outcomes. Overall, AOFAS score was improved from 41.3±11.3 points (range, 20-56 points) preoperatively to 85.5±7.1 points (range, 68-100 points) postoperatively (Z=5.16, P< 0.001), and the VAS score was reduced from 5.6±1.5 points (range, 3-8 points) preoperatively to 0.8±1.2 points (range, 0-4 points) postoperatively (Z=5.19, P< 0.001). These differences of functional score and pain evaluation were statistically significant. The Tomeno-Méary angle decreased from-6.6°±12.2° to 0.5°±2.9° (Z=3.43, P=0.001); the calcaneal pitch angle increased from 13.5°±3.9° to 22.1°±3.4°(t=10.12, P< 0.001), and the talarnavic-ular coverage angle decreased from 27.1°±5.9° to 7.6°±3.3°(Z=5.16, P< 0.001). The AP talar-first metatarsal angle decreased from-16.0°±10.7° to-7.0°±8.5°(t=5.49, P< 0.001). The differences of radiological parameters were statistically significant. Conclusion The mid-term results showed that the TN or TNC joint fusion could achieve a favorable clinical and radiological outcome for MWD patients. Even for the patients with severe deformities (stage II-V), such treatment strategy could also achieve satisfacto-ry deformity correction and functional improvement. Intraoperative restoration of talus rotation is the key to normal alignment of the subtalar joint/TN joint.

Neurogenic bowel dysfunction (NBD) manifested as constipation and fecal incontinence often occurs after spinal cord injury (SCI).NBD affects patients' quality of life and is an urgent clinical problem to be solved.The mechanism of NBD is related to central and autonomic nervous system dysfunction,intestinal nervous system dysfunction,changes in intestinal microorganism composition and abnormal content of neurotransmitters.The evaluation method of NBD is mainly based on scoring and imaging,which lacks unified criteria,and the treatment method for NBD is the combination of traditional Chinese and Western medicine.The author summarizes the mechanism,evaluation method,treatment and nursing of NBD in order to provide new insight into these aspects to improve clinical efficacy.

Objective: To analyze the clinical response in cases of multi-victims bitten by a dog in Beijing (one of whom was HIV positive), and to explore the route and prognosis of dual exposure to rabies virus and HIV. Methods: Descriptive epidemiological methods were used to analyze the exposed cases, the post-exposure prophylaxis(PEP) of rabies and HIV and the follow-up outcomes. Results: After six months-follow up, there was no rabies case was found among those nine victims bitten by the suspected dog. In addition, six HIV-negative victims who had been exposed to HIV via dog saliva were consistently tested to be HIV negative in the follow up period. Conclusions In cases with multi-victims bitten by one dog, except exposure to rabies virus, it is necessary to be alert to the presence of HIV, HBV or HCV infection in sequential victims. Therefore, the risk of the potential spreading of other blood-borne infectious pathogens should be assessed. It is important to initiate PEP as early as possible.

Objective: To understand the prophylaxis of population exposed to rabies, and provide a basis for prevention and control of rabies. Methods: The registration data of 1 019 cases with rabies exposure in Beijing Ditan Hospital Affiliated to Capital Medical University from Mar, 2017 to Feb, 2018 were analyzed using the descriptive epidemiologic method. Results: Among the 1 019 cases of rabies exposure, the sex ratio of men to women was 0.90∶1, with the highest proportion of them were between 26 and 46 years of age old, accounting for 46.5%. Rabies exposure reached its peak (46.9%) between June and September. In addition, 63.3% of the injuries were caused by dogs and 36.7% by cats. Most of the wounds (55.1%) occurred in hands; and the second was lower limbs (26.6%). Most of the cases (60.7%) had grade II wounds; followed by grade III wounds (39.1%); 69.3% of the cases completed immunization with rabies vaccine. In grade III exposure, 73.7% of the patients were immunized with rabies immunoglobulin. Most of the persons of grade III exposure (73.7%) received inoculation with human rabies immunoglobulin (HRIG). Conclusions Measures to control rabies should be focused on the management of dogs and cats and the standardization of rabies post-exposure prophylaxis.