ObjectiveTo determine the syndrome of a rat model of follicular dysplasia induced by Tripterygium glycosides based on prescriptions and investigate the mechanism of traditional Chinese medicine intervention via the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/hypoxia-inducible factor-1 （HIF-1）/vascular endothelial growth factor （VEGF） pathway. MethodForty-eight rats with regular estrous cycles were randomly assigned into a normal group （n=8） and a modeling group （n=40）. The rats in the modeling group were administrated with Tripterygium glycoside suspension （75 mL·kg^-1） by gavage for 30 days. The modeled rats were assigned into model， Siwutang （3.69 g·kg^-1）， Youguiyin （3.11 g·kg^-1）， Zuoguiyin （7.29 g·kg^-1）， and Guishenwan （10.35 g·kg^-1） groups， with 8 rats in each group. The drug intervention lasted for 14 days. The changes of estrous cycle were detected by Pap staining， and a stereoscope was used to observe the morphology of the ovarian tissue. Hematoxylin-eosin staining was employed to observe the pathological changes and follicle count in the ovarian tissue. Enzyme-related immunosorbent assay （ELISA） was used to measure the levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E₂） in the serum. Real-time fluorescence quantitative polymerase chain reaction and Western blot were employed to determine the mRNA and protein levels， respectively， of AMPK， mTOR， HIF-1， and VEGF in the ovarian tissue. ResultCompared with the normal group， the model group had a disordered estrous cycle， reduced secondary and mature follicles， increased atretic follicles， elevated FSH and LH levels， lowered E₂ level， up-regulated mRNA and protein levels of AMPK， and down-regulated mRNA and protein levels of mTOR， HIF-1， and VEGF （P<0.01）. Compared with the model group， Guishenwan increased secondary and mature follicles， decreased atretic follicles， lowered the FSH and LH levels， elevated the E₂ level， down-regulated the mRNA and protein levels of AMPK， and up-regulated the mRNA and protein levels of mTOR， HIF-1， and VEGF （P<0.01）. Compared with Guishenwan group， Siwutang， Youguiyin， and Zuoguiyin decreased mature follicles， increased atretic follicles （P<0.01）， elevated the LH （P<0.01） and FSH （P<0.05） levels， and lowered the E₂ level （P<0.05）. In addition， Youguiyin up-regulated the protein level of AMPK （P<0.05） and down-regulated the mRNA levels of mTOR and HIF-1 （P<0.01） as well as the mRNA and protein levels of VEGF （P<0.01）. Siwutang down-regulated the mRNA levels of mTOR and HIF-1 as well as the mRNA and protein levels of VEGF （P<0.05）. Zuoguiyin down-regulated the mRNA level of mTOR and the protein and mRNA levels of VEGF （P<0.05）. ConclusionGuishenwan may improve the ovarian function and promote follicle maturation in a rat model of follicular dysplasia by inhibiting the AMPK/mTOR/HIF-1/VEGF pathway， with the therapeutic effect superior to Zuoguiyin， Youguiyin， and Siwutang. It was hypothesized that this model presented the syndrome of kidney-essence deficiency.

Objective To study the effect of Ditan Decoction combined with aripiprazole and olanzapine in the treatment of schizophrenia and its influence on serum inflammatory factors chnage.Methods Seventy-seven patients with schizophrenia meeting the requirements visiting the outpatient department and hospitalized in Dazu District Hospital of Traditional Chinese Medicine and Dazu District Mental Health Center from July 2021 to March 2023 were selected as the study subjects and divided into the observation group(n=38)and control group(n=39).The control group was treated with aripiprazole and olanzapine,and the observation group was combined with Ditan Decoction on the basis of the control group.After 8 weeks of treatment,the TCM syndrome scores,Positive and Negative Syndrome Scale(PANSS)score,serum inflammatory factors(IL-6,IL-1β,IL-17)levels were compared between the two groups.Results The total effective rate was 97.37％in the observation group and 84.65％in the control group,and the difference was statistically signifi-cant(P＜0.05).The TCM syndrome score of each item and total scores after treatment in the observation group were lower than those in the control group(P＜0.05),the PANSS positive symptoms,negative symp-toms,general psychopathology and total scores in the observation group were lower than those in the control group(P＜0.05).The IL-17,IL-6 and IL-1β levels after treatment in observation group were lower than those in the control group(P＜0.05).Conclusion Ditan Decoction combined with aripiprazole and olanzapine has significant clinical efficacy in the treatment of schizophrenia,which could further reduce the symptom score of the patients and improve the serum inflammatory factors levels.The treatment is highly safe and worthy of clinical recommendation.

Objective:To investigate the clinical efficacy of the vascularised semi-split fibular flap with lateral approach in ankle fusion.Methods:A total of 54 patients who underwent ankle fusion through the lateral menstrual approach by the Department of Trauma and Micro Orthopaedics, Zhongnan Hospital of Wuhan University from June 2015 to December 2020 was retrospectively analysed. Of the 54 patients, 27 patients who underwent ankle fusion with a semi-split fibular flap carrying blood supply were assigned to the fibular flap group, while other 27 patients who had ankle fusion with lateral plate were assigned to the plate group. The ankle fusions for both groups were performed by the same surgical team. The clinical efficacy of the 2 groups was evaluated by the success rate of bone fusion, the American Orthopedic Foot and Ankle Surgery(AOFAS) Ankle-Hindfoot Score System, and the Visual analog score(VAS). All the patients entered the postoperative follow-up at outpatient clinic.Results:The follow-up lasted for 12-24 months, with an average of 15 months. At 6 months after surgery, 24 patients in the fibular flap group achieved osseous union with 88.9% in the success rate of bone fusion. While 17 patients in the plate group achieved osseous union, with the success rate of bone fusion at 63.0%. There was a statistically significant difference between the groups( P<0.05). At 1 year after surgery, all 27 patients in the fibular flap group achieved bony union, with a 100% of the success rate of bone fusion, while 23 patients in the plate group achieved bony union at a 85.2% success rate of bone fusion, with 4 patients failed to heal. The difference was statistically significant( P<0.05). The AOFAS scores of the fibular flap group were 41.3±12.0, 65.6±5.6, and 79.1±7.0 before operation, at 6 months after operation and 1 year after operation, respectively, while the scores for the plate group at the same time were 40.8±11.3, 64.5±4.1 and 69.3±7.2. There was no significant difference in the scores between the groups before surgery and at 6 months after the operation ( P>0.05). The scores at 1 year after surgery in the fibular flap group was significantly higher than the plate group, and there was a statistically significant difference( P<0.05). The preoperative VAS score in the fibular flap group was 7.6±1.3, while it was 7.5±1.1 in the plate group. There was no significant difference between the groups in VAS score( P>0.05). However, the VAS score at 1 year after surgery was found at 1.8±0.9 in the plate group, and 1.9±0.8 in the fibular flap group and the difference between groups was statistically significant( P<0.05). Conclusion:The surgical procedure of semi-split fibular flap through the lateral approach has lower complications and higher success rate in ankle fusion in comparison with those of the fusion with lateral plate. Thus, further clinical investigations can be considered.

Objective:To establish a method of serum detection by Raman spectroscopy for the diagnosis of gastric cancer.Methods:Between April and November 2019, 110 patients with gastric cancer [73 males, 37 females, age (57.4±10.3) years] and 74 patients with colorectal cancer [48 males and 26 females, aged (58.3±12.2) years] were collected at the First Affiliated Hospital of Army Military Medical University, along with 100 healthy subjects [59 males and 41 females, aged (55.6±10.61) years] during the same period. Fasting venous blood serum samples were collected from the subjects. A Raman spectrometer XploRA PLUS was used in this experiment, with an excitation light source of 532 nm, a field of view of 100 times, and a spectrum range of 200-2 000 cm -1, etc. The serum samples were detected by nondestructive and non-contact rapid detection, and the Raman spectra of serum samples were collected. Using the Raman spectrum acquisition and processing supporting software LabSpec6 to smooth, baseline, and normalize the obtained Raman spectrum. Multivariate statistical analysis software SIMCA14.1 were applied to import and analyze the obtained Raman spectrum data by principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and other methods for statistical analysis. An operating characteristic curve (ROC) was constructed to evaluate the model analysis effect between serum samples of healthy people and those with gastric cancer. Serum samples from the colorectal cancer group were used to verify the reliability of the model. Results:Six Raman peaks with good repeatability were detected in serum samples in health and gastric cancer group, and peaks were located at 1 001.17, 1 154.63, 1 337.89, 1 446.85, 1 515.33, and 1 658.34 cm -1, respectively. Raman intensities at six Raman peaks were significantly different between healthy and gastric cancer groups. At the displacement of 1 001.17, 1 154.63, and 1 515.33 cm -1, the Raman intensity in the healthy group was higher than that in the gastric cancer group. At 1 337.89, 1 446.85, and 1 658.34 cm -1 displacement, the Raman intensity of the gastric cancer group was higher than that of the healthy group. An OPLS-DA model was constructed to analyze the serum samples of the healthy group and the gastric cancer group. In the model, R 2 is the fitting power, and Q 2 is the predictive ability. The closer the values of R 2 and Q 2 are to 1, the better the performance of the model, and the obtained model's R 2X(cum)=0.809, R 2Y(cum)=0.819, Q 2(cum)=0.758. ROC characteristic curve was drawn based on the OPLS-DA model. The area under the curve (AUC) of the gastric cancer group was 0.998. Six peaks with good repeatability were detected in the serum Raman spectra of gastric cancer stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ, which were located at the displacement of 1 001.85, 1 155.07, 1 338.36, 1 445.75, 1 515.92, and 1 657.68 cm -1, respectively, and at the displacement of 1 155.07 and 1 515.92 cm -1. The Raman intensity of gastric cancer stage Ⅳwas significantly higher than that of gastric cancer stages Ⅰ, Ⅱ, and Ⅲ. Conclusions:According to the model reliability verification, the healthy group, gastric cancer group and colorectal cancer group can also be effectively separated based on OPLS-DA results; it showed a good performance in separating the healthy group from the gastric cancer group. It is possible to detect serum samples from healthy people and gastric cancer patients unlabeled by combining Raman spectroscopy and the OPLS-DA method in multivariate statistics.

Objective To investigate the surgical outcomes of one-stage repair of the ankle fracture combined with grade Ⅲ injury to the lateral ligament.Methods From April 2015 to December 2017,23 patients with ankle fracture and lateral ligament injury were treated at Department of Traumatic Reconstruction Surgery/Foot and Ankle Surgery,Zhongnan Hospital.There were 15 males and 8 females,aged from 18 to 65 years (average,37.8 years).After intraoperative treatment of their fractures,22 ruptures of the ligamentous insertion were repaired with suture anchor and one rupture of the middle ligament with Internal Brace.Routine postoperative X-rays,physical examination and Sefton grading system were used to evaluate the stability of the ankle joint and the subtalar joint.The Karlsson-Peterson and American Orthopedic Foot and Ankle Society (AOFAS) scores were used to evaluate the function and pain of the ankle.Results The follow-up for this cohort ranged from 12 to 24 months (average,13.9 months).The bone union time ranged from 6 to 24 weeks,averaging 10.1 weeks.There was no chronic instability of lateral ankle or subtalar joint in all the cases.Two cases developed osteochondral lesion of the medial talus which was treated with micro-fracture surgery one year after injury;one case suffered subtalar arthritis which was treated by subtalar arthrodesis 14 months after injury.Their Karlsson-Peterson scores averaged 84.6 points and AOFAS ankle-hindfoot scores 93.7 points with 19 excellent and 4 good cases.The Sefton grading system gave 9 cases of grade Ⅰ and 14 ones of grade Ⅱ.Conclusion One-stage repair of the ankle fracture combined with grade Ⅲ injury to the lateral ligament can lead to the stability of the ankle joint and the subtalar joint and improved function of the ankle.

Objective To investigate the clinical effectivity of the muscular flap transposition and induced membrane technique in the emergency treatment for the limb salvage of Gustilo type Ⅲ B/C open fracture of lower leg. Methods From July, 2015 to December, 2017, 10 cases of Gustilo type Ⅲ B/C fracture of lower leg with bone defects were performed limb salvage surgery. Induced membrane technique was used to fill the bone defects in the emergency room.The gastrocnemius and/or soleus muscular flaps were transposed to cover the bone cement or ex-posed bone simultaneously in emergence treatment. After the wound healed completely, traditional bone grafting was used to repair the bone defects. There were 4 cases of Gustilo type Ⅲ B and 6 cases of Gustilo type Ⅲ C. The aver-age length of bone defect was (5.25±1.70) cm ranging from 3.0 cm to 11.0 cm. The gastrocnemius medial head flaps were performed in 5 cases, the combined application with the gastrocnemius medial head flaps and the medial hemimuscular flaps of soleus were performed in 2 cases, and medial hemimuscular flaps of soleus were transposed in 3 cases. Results The wounds in 6 cases were healed at one stage, but 2 cases healed by dressing because the exudate after skin grafting.In 1 case, the cross-leg flap was used to cover the exposed bone cement due to the necro-sis of soleus flap. The other 1 was performed the transposition of the lateral gastrocnemius flaps because the exposure of bone cement after the necrosis of the upper and lateral muscles in lower leg. In the second-stage, the bone defects were reconstructed by traditional bone grafting. The average healed time of bone was 7.2 months ranging from 5 months to 9 months. At the last followed-up time, all patients recovered their function of weight-bearing. The Paley's score of the adjacent joints: excellent in 8 cases and good in 2 cases. Conclusion The combination with induced membrane technique and local muscular flap transposition in emergency surgery is an effective method to limb salvage for the Gustilo type Ⅲ B/C open fracture of lower leg.

Objective: To observe the influence of TRPV6 gene silence on SW480 colon cancer cell biological behavior, change in the in-tracellular concentration of calcium, as well as influence of 1,25 (OH)2D3CaCl2and CuCl2on SD rat colonic neoplasm models. Method: SW480 colon cancer cells were infected using lentivirus particles. TRPV6 protein and mRNA expression was detected using immunohistochemical tests, Western blot, and PCR. Moreover, the proliferation, metastasis, and apoptosis of SW480 colon cancer cells were detected through MTT assay and metastasis and apoptosis experiments, and the concentration of Ca2+in SW480 colon cancer cells was measured using high-speed ionic imaging. The SD rat colon cancer model was established based on DMH, and were assigned into experimental group (DMH group, 15) and intervention group (DMH+1,25 (OH)2D3group, DMH+CuCl2group) and control group, 10 in each group. The SD rat colon cancer model is established based on DMH, given 1,25(OH)2D3(37.5 nmol/kg) and CuCl2(375 μmol/kg) separately as intervention. The occurrence of colonic neoplasms and glandular cancers in each group of rats was observed, and Western blot was employed for detection of the TRPV6 protein expression. Results: After the transfection of SW480 colon cancer cells by TRPV6-RNAi, the expression of TRPV6 mRNA and protein decreased, intracellular concentration of Ca2+decreased, proliferation and metastasis rate of SW480 colon cancer cells decreased, and apoptosis rate of these cells increased. The differences between the groups with intervention and the blank control group and negative control group showed statistical significance (P<0.05). The colon cancer occurrence rate in the control group was 0, while that of the DMH+1,25 (OH)2D3 group, DMH group, and DMH+CuCl2were 100%, 84.62%, and 33.33%, respectively. The TRPV6 protein expression was detected in all groups, while DMH+1,25(OH)2D3group was observed to exhibit the highest level of expression, followed by the DMH group, DMH+CuCl2group, and control group. The differences were of statistical significance (P<0.05). Conclusions: The proliferation and metastasis of SW480 colon cancer cells can be prohibited by lowering the concentration of Ca2+in the cells. Thus, the apoptosis of the cells can be induced. 1,25 (OH)2D3can help improve the expression of TRPV6 protein in experimental rat colon tissues and promote the formation of colon neoplasms. CuCl2can help lower the expression of TRPV6 protein in experimental rat colon tissues and prevent the formation of colon neoplasms.

Objective To investigate the clinical efficacy of three-stage Masquelet technique in the treatment of infective bone defects of foot and ankle.Methods A retrospective case series study was conducted on the clinical data of 19 patients with infective bone defects of foot and ankle admitted to Zhongnan Hospital of Wuhan University from December 2014 to October 2017.There were 15 males and four females,aged 18-68 years [(39.6 ± 12.3)years].Among the patients,16 patients were infected with bacteria and three patients were infected with Mycobacterium tuberculosis.The infection involved humeral end,talus and surrounding joints in 14 patients,internal hemorrhoids in two patients,midfoot and interphalangeal joints in one patient,and humerus and metatarsophalangeal joints in two.The operation included three stages:the first stage was thorough debridement,supplemented with negative pressure closed drainage (VSD) and continuous washing to clarify the pathogenic bacteria;the second stage was to fill the bone defect with targeted antibiotic bone cement to prevent or treat infection;in the third stage,after filling the antibiotic bone cement for 3 months with no sign of local wound infection,the bone cement was taken out,and the bone reconstruction operation was performed by means of internal fixation and bone grafting.The flap survival and wound healing were observed,and the time of fracture healing was recorded.The American Orthopaedic Foot and Ankle Society (AOFAS) score and the Visual Analogue Score (VAS) were used to evaluate the improvement of the function of the foot and ankle before operation and at the last follow-up,and the bone healing was evaluated according to the radiographic union scale in tibial (RUST) fractures.Results The patients were followed up for 9-12 months [(11.1 ±1.0) months].Two patients with soft tissue defects caused by preoperative infection and necrosis received posterior tibial artery perforator flap and anterolateral thigh flap repair in the second stage,and the flaps all survived.The postoperative bone healing time was 3 7 months [(3.5 ± 1.4)months].Nineteen patients underwent three-stage surgery,and the ankle and foot wound or sinus tract were all healed,with no infection recurrence during follow-up.At the last follow-up,the AOFAS score was improved significantly from preoperative (36.3 s-12.1) points to (71.4 ± 5.7) points (P ＜ 0.05).The VAS was decreased significantly from preoperative (5.3 ± 1.2) points to (1.4 ± 0.9) points (P ＜ 0.05).The RUST bone healing score at the last follow-up was 8-12 points [(10.2 ± 1.1) points].Conclusion In treating the infective bone defects of foot and ankle,the three-stage Masquelet technique can effectively control infection,facilitate wound healing,promote bone union,and improve foot and ankle function.

Objective:To investigate the role of TRPV5 and TRPV6 in intracellular calcium regulation and biological behaviors of SW480 colon cancer cells. Methods:qRT-PCR, Western blot, and immunocytochemistry were applied to determine the mRNA and protein ex-pression levels of TRPV5 and TRPV6 in SW480 colon cancer cell line upon treatment with TRPV5 and TRPV6 agonist, 1-25(OH)2D3, and inhibitor, CuCl2. The change of intracellular Ca2+level was examined with a confocal laser scanning microscope. Scratch test, MTT, and TUNEL assays were used to analyze the cell migration, proliferation, and apoptosis, respectively. Results:As an agonist of TRPV5 and TRPV6, 1-25(OH)2D3 significantly up-regulated the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 cell lines. On the other hand, CuCl2, being an inhibitor of TRPV5 and TRPV6, effectively down-regulated the TRPV5 and TRPV6 mRNA and protein expres-sion levels (P<0.05). The intracellular calcium concentration in SW480 cell line significantly increased upon treatment with 1-25 (OH)2D3, and significantly decreased with CuCl2 treatment (P<0.05). 1-25(OH)2D3 promoted cell proliferation and migration, and inhibit-ed apoptosis of SW480 cell in a time-and dose-dependent manner (P<0.05). However, CuCl2 significantly repressed cell proliferation and migration and induced apoptosis (P<0.05). Conclusion: TRPV5 and TRPV6 can affect the biological behaviors of colon cancer SW480 cells by regulating intracellular Ca2+level.

Background and purpose:Previous studies have suggested Na+-Ca2+ exchanger isoform 1 (NCX1) as a key component of calcium homeostasis was involved in the tumorigenesis. However, the role of NCX1 and calcium signal in tumorigenesis of hepatocellular carcinoma (HCC) has not been explored. This study aimed to investigate the effect of NCX1 on cell proliferation and migration of HCC HepG2 cells in vitro and the possible mechanism.Methods:Both the real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were applied to assess the expression of NCX1 mRNA and protein in normal hepatic cells (LO2), HCC cell line (HepG2), human normal hepatic tissues and hepatocellular carcinoma tissues. The change of intracellular calcium signal in LO2 and HepG2 cells via acti-vated NCX1 channel in the presence or absence of Na+ was examined by a confocal laser scanning microscope. The effects of NCX1 special inhibitor KB-R7943 on cell proliferation and migration of HepG2 cells were measured by MTT and cellscratch test.Results:Both mRNA and protein expression of NCX1 were higher in HCC tissues and cell line HepG2 than in the normal tissues and cell line LO2 (P<0.05). The activation of NCX1 channel induced a slight rise in cytoplasmic Ca2+concentration ([Ca2+]cyt) in normal cells, but caused a marked increase in cancer cells. And the NCX1 activation induced intracellular calcium increase was significantly reversed by NCX1 inhibitor KB-R7943 (P<0.05). Both NCX1-mediated proliferation and migration of HepG2 were also significantly attenuated by the KB-R7943 (P<0.05).Conclusion:NCX1 is up-regulated in HCC cells and tissues. The activation of NCX1 mediates intracellular calcium homeostasis. The inhibition of NCX1 activity can suppress the proliferation and migration of HepG2 cells. It is suggested that NCX1 may be involved in the development and progression of HCC.