C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

Objective To investigate the efficiency of Trichostatin A (TSA) in inducing cell apoptosis and altering the Notch pathway genes expression in PANC-1 cells line.Methods The survival rate and apoptosis of PANC-1 cells were measured by MTT assay and Hoechst 33258 staining,respectively.mRNA expression levels of the genes,numb,gcn512,dll3,hes6,eaf2,cytohesins,in PANC-1 cells were assessed by real-time quantitive PCR.Western blot was used to measure the expression of bcl-2,bax,actived caspase-3 and NICD protein which was the biologically active form of Notch-1.Results After culturing with 0.1,0.2,and 0.4 μmol/L TSA for 24 hours,the cellular survival rate of PANC-1 cells significantly decreased to 72％,58％ and 39％,respectively.The survival rate of PANC-1 was negatively correlated to time length of culture with TSA.Increased apoptosis of PANC-1 cells after 12,24 and 36 h culture with TSA was detected by Hoechst 33258 staining.Western blotting showed that the expression of bax,actived caspase-3 and NICD protein increased while the bcl-2 protein decreased after culture with TSA.In real time quantitive PCR assessment,the mRNA expression of numb and hes6 in PANC-1 cells were upregulated by TSA (P ＜ 0.05),while the mRNA expression of gcn512 and dll3 were down-regulated by TSA (P ＜ 0.05).While mRNA expressions of eaf2 and cytohesin1,2,3,4 were not affected by TSA.Conclusions TSA induces apoptosis of pancreatic cancer cell line PANC-1.The Notch signal pathway may be involved in inducing cellular apoptosis of PANC-1 when cultured with TSA.

Some unhealthy life habits, such as long-term smoking, heavy drinking, sexual overstrain and frequent stay-up could induce the Yin deficiency symptoms of zygomatic red and dysphoria. Stems of Dendrobii officinalis flos (DOF) showed the efficacy of nourishing Yin. In this study, the hyperthyroidism Yin deficiency model was set up to study the yin nourishing effect and action mechanism of DOF, in order to provide the pharmacological basis for developing DOF resources and decreasing resource wastes. ICR mice were divided into five groups: the normal control group, the model control group, the positive control group and DOF extract groups (6.4 g · kg(-1)). Except for the normal group, the other groups were administrated with thyroxine for 30 d to set up the hyperthyroidism yin deficiency model. At the same time, the other groups were administrated with the corresponding drugs for 30 d. After administration for 4 weeks, the signs (facial temperature, pain domain, heart rate and autonomic activity) in mice were measured, and the facial and ear micro-circulation blood flow were detected by laser Doppler technology. After the last administration, all mice were fasted for 12 hours, blood were collected from their orbits, and serum were separated to detect AST, ALT, TG and TP by the automatic biochemistry analyzer and test T3, T4 and TSH levels by ELISA. (1) Compared with the normal control group, the model control group showed significant increases in facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01), serum AST, ALT (P < 0.01), T3 level (P < 0.05), TSH level (P < 0.05) and notable deceases in pain domain (P < 0.01), TG level (P < 0.01). (2) Compared with the model control group, extracts from DOF (6 g · kg(-1)) could notably reduce facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01) and AST (P < 0.05) and enhance pain domain (P < 0.01) and TG (P < 0.01). Extracts from DOF (4 g · kg(-1)) could remarkably reduce AST and ALT levels (P < 0.01, 0.05). Extracts from DOF (6 g · kg(-1) 4 g · kg(-1)) could significantly reduce T3 and increase serum TSH level (P < 0.05). DOF could improve Yin deficiency symptoms of zygomatic red and dysphoria in mice as well as liver function injury caused by overactive thyroid axis. According to its action mechanism, DOF may show yin nourishing and hepatic protective effects by impacting thyroxin substance metabolism, improving micro-circulation and reducing heart rate.
Animals ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flowers ; chemistry ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Thyroxine ; metabolism ; Yin Deficiency ; drug therapy ; metabolism

As the main chemical component of aromatic traditional Chinese medicine (TCM), the volatile oil of TCM has significant pharmacological effects, such as antibacterial, anti-inflammatory, antioxidant and so on. However, TCM volatile oil is easy to volatilize and oxidize, which seriously limits its application. As a kind of grid structure material, porous material has the characteristics of high specific surface area, large pore volume, adjustable pore size, strong adsorption capacity and controllable surface chemical properties. It has been widely used in adsorption separation, biomedicine, industrial catalysis, wastewater treatment and other fields. In recent years, the use of porous materials to adsorb volatile oil has provided a new strategy and method for improving the stability of TCM volatile oils. At the same time, it can realize the solidification and stability of TCM volatile oils and the application of preparations. In this review, the development and characteristics of porous materials such as mesoporous silica, mesoporous carbon, mesoporous nano hydroxyapatite, porous metal organic framework, porous starch and their application in improving the stability of TCM volatile oils are summarized, and the research strategies affecting the adsorption stability of porous materials for TCM volatile oils are discussed, in order to provide reference for the stabilization control and application of TCM volatile oils.

OBJECTIVE To estimate the value of video-assisted thoracoscopic surgery (VATS) in diagnosis and treatment of children with chest diseases. METHODS From May 1997 to October 2001, forty-one children (25 boys and 16 girls) with chest diseases received VATS under general anesthesia in our hospital. Their average age was 6.9 years (range 9 days to 16 years) and their median body weight was 22.5 kg (2.8-54.0 kg). Operative procedures included fibrinopurulent empyema with debridement and decortication in 15 children, biopsy and(or) resection of mediastinal tumor in 11, bullectomy and cystectomy of the lung in 6, lobectomy with huge cyst of the lung or sequestration in 5, clearance of hemothorax in 2, and exploration, and repair of diaphragmatic hernia in 2. RESULTS The mean operative time was 74 minutes (range 30 to 220 minutes). The lost blood volume was 33 ml (range 10 to 150 ml). The mean duration of chest drainage and hospital stay after surgery was 2.4 days and 7.0 days (range 4 to 15 days) respectively. One infant born after 9 days with congenital diaphragmatocele died of respiratory failure due to left pulmonary hypoplasia 10 days after operation. Postoperative morbidity was 7.3% (3 patients). Forty patients were followed up for an average of 15.6 months, 38 patients lived and developed normally, and 2 received chemotherapy. CONCLUSION Video-assisted thoracoscopy is a safe and effective diagnostic and therapeutic procedure for children with chest disease, and this approach has an important place in pediatric thoracic surgery.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Postoperative Complications ; epidemiology ; Thoracic Surgery, Video-Assisted ; adverse effects ; methods

OBJECTIVETo assess the clinical value of three-dimensional contrast-enhanced magnetic resonance angiography (3D-CE-MRA) in the diagnosis of mesenteric arteriosclerosis.

METHODS3D-CE-MRA of the mesenteric arteries was performed in 21 patients with 23 healthy subjects as the control. After 3D image reconstruction and maxi intense projection, and the abnormalities of the mesenteric arteries were observed and analyzed. The diameter and number of the arterial branches were compared between the patients and the control subjects.

RESULTSAll the 21 patients suffered arteriosclerosis in the arteries other than the mesenteric arteries. On 3D-CE-MRA, mesenteric arteriosclerosis was characterized by thinning of the arterial trunk, luminal stenosis, irregular arterial margins and homo- or heterogeneous thickening of vascular walls. Multiple filling defects were found in the mesenteric artery lumens with reduced second order branches, which showed rigid lining with dashed line appearance or disappeared in some cases. The inferior mesenteric arteries were seen in only 2 patients. The diameters of superior and inferior mesenteric arteries were 3.8-/+0.32 mm and 1.20-/+0.12 mm in the patients, significantly smaller than those of in the control subjects (6.51-/+1.01 mm and 2.90-/+0.90 mm, respectively, P<0.01). The number of the mesenteric artery branch of the patients was also significantly reduced as compared with that in the control subjects (P<0.05). In som cases, the intestinal enhancement was attenuated with the intestinal contraction, dilatation and lowering of the intestinal tension.

CONCLUSION3D-CE-MRA can clearly display mesenteric arteriosclerosis and secondary intestinal changes, and provides a useful means for the diagnosis and assisting the therapy of mesenteric arteriosclerosis.

Aged ; Aged, 80 and over ; Arteriosclerosis ; diagnosis ; pathology ; Case-Control Studies ; Contrast Media ; Female ; Humans ; Image Enhancement ; Imaging, Three-Dimensional ; Magnetic Resonance Angiography ; methods ; Male ; Mesenteric Arteries ; pathology ; Middle Aged

Objective To describe the epidemic status of hepatitis B along the Yangzi River area,ill Anhui province.Methods A croSS-sectional seroepidemiological survey was conducted and the samples were collected by stratified cluster sampling.Serological biomarkers to hepatitis B virus were tested using enzyme-linked immunosorbent assay(ELISA)reagents.Results Among 2282 people.the prevalence rates of HBsAg,HBsAb,HBeAg.HBeAb,HBeAb and hepatitis B virus infection were 9.8%,43.5%.1.0%,7.8%,10.3%and 40.6%respectively.The prevalence rate of HBsAg among males was higllel"than that offemales(P＜0.05).The vaccination rate was 24.7%,higher in urban than in taral areas.The vaccination rate was high in children younger than ten years old and in students.The prevalence rates of HBsAg and HBV among people who had received vaccines were lower than those who had not.Conclusion The standardized prevalence rates of HBsAg(9.3%)Was high in the area along the Yangzi River in Althai province.The vaccinate rate Was low in the country side.Expanded vaccinate which can obviously reduce the prevalence rate of HBsAg should be enhanced.

Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.

Objective: To explore the effects of autologous platelet-rich plasma (PRP) in the repair of soft tissue defects of rabbits with free flap. Methods: Thirty 6-month-old New Zealand white rabbits, male and female unlimited, were used to harvest blood from the heart. PRP was prepared by Aghaloo method, then free flap model with size of 5 cm×3 cm was reproduced on each ear of the rabbit. According to the random number table, one ear of each rabbit was recruited to PRP group, and the other ear was recruited to normal saline group. The base of flap on rabbit ear in PRP group was evenly spread with 1.0 mL autologous PRP, and equivalent volume of normal saline was applied to that in normal saline group. Then, the flap was replanted in situ. On post surgery day (PSD) 2, 3, 5, 7, and 14, 6 rabbits in each group were taken. The survival of flap was observed and recorded. The morphology of the basal tissue of flap was observed by hematoxylin-eosin staining. The expressions of CD31 and α smooth muscle actin (α-SMA) in the basal tissue of flap were detected by immunofluorescence method. Another 6-month-old male New Zealand white rabbit without making flap under the same experimental conditions was used for harvesting whole blood and preparing PRP. Then blood platelet count in whole blood and PRP was determined, and the content of vascular endothelial growth factor (VEGF) and transforming growth factor β (TGF-β) was detected by double-antibody sandwich enzyme-linked immunosorbent assay. Data were processed with analysis of variance of factorial design, paired sample t test, and Bonferroni correction. Results: (1) On PSD 2, the flaps of wounds of rabbits in PRP group were reddish and adhered well to the basal tissue; the flaps of wounds of rabbits in normal saline group were dark red and poorly attached to the basal tissue. On PSD 3, the flaps of wounds of rabbits in PRP group were ruddy and closely adhered to the basal tissue; the flaps of wounds of rabbits in normal saline group were scattered in the plaque-like dark red and generally attached to the base. On PSD 5, the flaps of wounds of rabbits in PRP group were reddish and closely adhered to the basal tissue, and the flaps were alive; while flaps of wounds of rabbits in normal saline group were rosy and closely adhered to the basal tissue. On PSD 7, the surface of flaps of wounds of rabbits in PRP group was covered with a medium amount of rabbit hair. The color of flap was similar to that of the surrounding skin. The flaps of wounds of rabbits in normal saline group were generally attached to the base, and the surface was only covered with a small amount of fluff. On PSD 14, the incisions were healed well in PRP group, while small wounds in normal saline group were not healed. (2) On PSD 2, inflammatory cell infiltration was observed in flaps of wounds of rabbits in both groups. On PSD 3, the flaps of wounds of rabbits in PRP group showed neovascularization, with less interstitial hemorrhage; while there were less neovascularization in the flaps of wounds of rabbits in normal saline group. On PSD 5, a medium number of inflammatory cell infiltration and a small amount of new microvessels were observed in flaps of wounds of rabbits in normal saline group. Many fibroblasts, a small amount of inflammatory cells, and scattered new microvessels were observed in flaps of wounds of rabbits in PRP group. On PSD 7, the number of new microvessels in normal saline group was significantly lower than that in PRP group. On PSD 14, the new microvessels in the flaps of wounds of rabbits in PRP group gradually matured, and a large number of fibroblasts distributed around them. Some of the newly formed microvessels in the flaps of wounds of rabbits in normal saline group were mature, and the healing was slower than that of PRP group. (3) On PSD 2, 3, 5, 7, and 14, the expressions of CD31 and α-SMA in the basal tissue of flaps of wounds of rabbits in PRP group were significantly higher than those in normal saline group (t=10.133, 5.444, 9.450, 6.986, 8.394, 14.896, 10.328, 9.295, 13.902, 10.814, P<0.01). (4) The platelet count in activated PRP of rabbits was (2 863±962)×10⁹/L, which was significantly higher than (393±49)×10⁹/L in whole blood (t=7.690, P<0.05). (5) The content of VEGF and TGF-β in activated PRP of rabbits was (564.3±3.2) and (1 143±251) pg/mL, which was significantly higher than (99.7±0.4) and (274±95) pg/mL in whole blood, respectively (t=287.390, 9.648, P<0.05 or P<0.01). Conclusions PRP of rabbits contains high concentrations of VEGF and TGF-β. Therefore, PRP can effectively promote microvascular regeneration in free flap tissue and accelerate the survival of free flap.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.