28 days after injury (P < 0.05). In the fracture+spinal cord injury group, the level of serum transforming growth factor beta 1 had a rapid increase on the 7th day, and reached the peak on the 14th day, and then, this level had no significant decrease until the 28th day. In the simple fracture group, the level of serum transforming growth factor beta 1 began to increase on the 2nd day, reached the peak on the 7th day, and then decreased gradualy. Remarkable changes of serum transforming growth factor beta 1 levels in patients with bone fracture combined with spinal cord injury may be associated with fracture healing in different periods.

BACKGROUND:The method in repair of primary lumbar intervertebral infection is different in different positions, mainly containing anterior, posterior and anteroposterior pathways. In posterior pathway, muscle gap approach is recognized by many fel ows. This program has its special advantage compared with conventional posteromedial approach. OBJECTIVE:To evaluate the clinical effects of debridement, bone graft and internal fixation of pedicle screw placement in repair of primary lumbar intervertebral infection through posterior paraspinal muscle approach. METHODS:Clinical data of 13 patients with primary lumbar intervertebral infection were analyzed retrospectively. There were one case of L 2-L 3 , two cases of L 3-L 4 , four cases of L 4-L 5 and six cases of L 5-S 1 . Lumbar pain was obvious in al patients. Nine cases suffered from radioactive lower limb pain. Al patients received debridement, bone graft and internal fixation of pedicle screw placement through paraspinal muscle approach via posteromedial incision. After treatment, clinical effects were evaluated using Visual Analog Scale and lumbar Japanese Orthopaedic Association scores. RESULTS AND CONCLUSION:Al patients were fol owed up for 12-18 months, no recurrence. X-ray review demonstrated bony fusion, so loosening and breakage were not found in al patients. Visual Analog Scale scores revealed that Visual Analog Scale score was 8.15 preoperatively, 2.15 at 1 week postoperatively, 1 at final fol ow-up, showing significant difference (P<0.05). There was significant difference in pain between pre-treatment and post-treatment, and pain apparently relieved after treatment. Lumbar Japanese Orthopaedic Association score showed that effective outcomes were found in al patients after treatment, including nine cases of excel ent effects, three cases of good effects, one case of average effects, with the excel ent and good rate of 92%. Above findings confirmed that one-stage debridement, bone graft and internal fixation of pedicle screw placement through posterior muscle gap approach provides a good repair method for patients with primary lumbar intervertebral infection. It can achieve intervertebral space directly through intervertebral foramen, retain the central spinous process and lamina, reduce the injury to paraspinal muscle, and keep spinal ligament complex. However, strong spine fixation contributes to bone fusion, and keeps the stability of the spine after repair.

Objective To explore the changing trend of Ia on monocyte, lymphocyte apoptosis rate, TNF-α and IL-6 in abdominal aorta of burned rats with delayed resuscitation and the influence of application of carbachol on them. Methods Adult male Wistar rats were randomly divided into normal control group(n =8), scald group(n =48) and scald with carbachol treatment group(n =48). In latter two groups, rats were inflicted with 30% total body surface area (TBSA) full-thickness scald and delayed fluid resuscitation. All scald rats were sacrificed at the 6th hours or 1st, 2nd, 3rd, 7th, 14th day after scald, with 8 rats at each time point. Expression of Ia antigen on monocyte and lymphocyte apoptosis rate were determined by direct immunofluorescence on a flow cytometer, and TNF-α and IL-6 was measured by ELISA. Results Expression of la on monocyte was obviously lower than that of controls. The lowest levels were recorded on the 6th hours and 1st day after scald. Subsequently, Ia was elevated gradually, but still lower than that of normal rats(P <0. 01). After administration of carbachol, Ia expression was obviously promoted, compared with the simple scald group (P <0. 01). Lymphocyte apoptosis rate, TNF-α and IL-6 was higher than that of controls(P <0. 01). After administration of cavachol, , lymphocyte apoptosis rate and TNF-α and IL-6 was obviously down-regulated on the 6th hours, 1st day, 2nd day and 3rd day after scald injury, compared with the simple scald group (P < 0. 01 or 0. 05). Conclusion After severe burn with delayed fluid resuscitation, there is a low la expression, high lymphocyte apoptosis rate and increased releasing of proinflammatory cytokine. Immune function was suppressed. Carbacho] could improve the immune function of scald rats with delayed fluid resuscitation.

COVID-19 epidemic continues to spread around the world till these days, and it is urgent to develop more safe and effective new drugs. Due to the limited P3 biosafety laboratories for directly screening inhibitors of virulent viruses with high infectivity, it is necessary to develop rapid and efficient screening methods for viral proteases and other related targets. The main protease (M^pro), which plays a key role in the replication cycle of SARS-CoV-2, is highly conserved and has no homologous proteases in humans, making it an ideal target for drug development. From two different levels, namely, molecular level and cellular level, this paper summarizes the reported screening methods of SARS-CoV-2 M^pro inhibitors through a variety of representative examples, expecting to provide references for further development of SARS-CoV-2 M^pro inhibitors.

As a common protease with high similarity among coronavirus species, the main protease (M^pro) of SARS-CoV-2 is responsible for the catalytic hydrolysis of viral precursor proteins into functional proteins, which is essential for coronavirus replication and is one of the ideal targets for the development of broad-spectrum antiviral drugs. This paper reviews the main protease inhibitors of SARS-CoV-2, including their molecular structures, potencies and drug-like profiles, binding modes and structure-activity relationships, etc.

BACKGROUNDEstrogen might play an important role in type 2 diabetes mellitus pathogenesis. A number of polymorphisms have been reported in the estrogen receptor alpha (ERalpha) gene (also named ESR1), including the XbaI and PvuII restriction enzyme polymorphisms of ESR1, which may be involved in disease pathogenesis. The aim of this study was to determine whether ERX gene polymorphisms are associated with type 2 diabetes mellitus and serum lipid level.

METHODSTwo hundred and ninety-nine patients with type 2 diabetes mellitus were compared with three hundred and forty-one health controls of Guangzhou in China, both were male and postmenopausal female residents at 51 - 70 years. ESR1 genotyping was performed using polymerase chain reaction (PCR) and PvuII and XbaI restriction fragment length polymorphism (PCR-RFLP) analysis.

RESULTSESR1 allelic frequencies of P, p and X, x alleles were 0.408, 0.592; 0.360, 0.640 in the type 2 diabetes mellitus group and 0.318, 0.682; 0.328, 0.672 in the control group, respectively. In case-control study, there was significant difference in PvuII, but not XbaI, allele frequency between the type 2 diabetes mellitus and control groups (P = 0.001 and P = 0.122). When the group was separated into men and women, the difference was significant in women (P < 0.001) but not in men (P = 0.854) with the PvuII genotype, and the effect of PvuII variant on the development of type 2 diabetes mellitus was improved with aging. In addition, PvuII genotype was associated with blood glucose [fasting blood glucose (FBG), postprandial blood glucose (PBG)] and serum lipid [total cholesterol (TC) and low density lipoprotein (LDL)-c] concentration in healthy women.

CONCLUSIONSPvuII polymorphism of ESR1 increases susceptibility to type 2 diabetes mellitus in Chinese Guangzhou women. ESR1 variants may also impact serum lipid metabolism, which might provide a mechanism connecting ESR1 to type 2 diabetes.

Aged ; Blood Glucose ; analysis ; Cholesterol, LDL ; blood ; Diabetes Mellitus, Type 2 ; blood ; genetics ; Estrogen Receptor alpha ; genetics ; Female ; Genotype ; Humans ; Lipids ; blood ; Logistic Models ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo investigate the effect of VEGF expression in osteosarcoma cell line and the target killing effect of HSV1-TK/GCV system on transfected osteosarcoma cells under hypoxia conditions.

METHODSEukaryotic expression plasmid with HRE promoter was constructed to express the antisense VEGF165 cDNA and Hygromycin phospho-transferase-thymidine kinase (HyTK) fusion gene. The recombinant vectors were then transfected into osteosarcoma cell line MG63 with lipofectin mediated gene transfer methods. PCR and RT-PCR were used to confirm the presence and expression of TK gene. The sensitivity of transfected cells to GCV and "bystander effect (BSE)" of HSV1-TK/GCV system under normoxia or hypoxia conditions were measured by MTT assay and mixed co-culture experiment. The expression of VEGF protein was detected by ELISA under hypoxia condition. Cell cycle phase distribution was determined by flow cytometry. In addition, electromicroscopy was used to document ultrastructural alterations.

RESULTSThe eukaryotic expression vector pBI-HRE-AsVEGF165 -HyTK was constructed successfully. The transfected cell line MG63TV was established and confirmed by PCR and RT-PCR of the presence of transgene and its mRNA expression. GCV was toxic to transfected cells in a concentration-dependent manner. The sensitivity to GCV toxicity was 100 times higher under hypoxia condition than that under normoxic condition. The mixed culture experiments showed that the "bystander effect" was enhanced significantly under hypoxia condition. VEGF expression of transgene cells under hypoxia condition decreased 50% compared to that of normal condition. Under hypoxia and GCV, DNA synthesis of MG63TV cells was inhibited along with an increase of cells at G0 approximately G1 phase, apoptosis and necrosis.

CONCLUSIONSAntisense VEGF expression driven by HRE promoter in combination with hypoxia can provide a target inhibition of VEGF expression in human osteosarcoma cells, with an enhanced selective killing effect and BSE of the HSV-TK/GCV system. The double-gene co-expression system in study provides experimental basis for therapy against osteosarcoma by a synchronous antiangiogenic and suicide gene approach.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Bystander Effect ; Cell Hypoxia ; Cell Line, Tumor ; Cell Proliferation ; DNA, Neoplasm ; biosynthesis ; Ganciclovir ; pharmacology ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1 ; genetics ; Oligodeoxyribonucleotides, Antisense ; Osteosarcoma ; metabolism ; pathology ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Thymidine Kinase ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVEUtilizing the hypoxia inducible factor 1/hypoxia reaction element (HIF-1/ HRE) gene regulation system to construct antisense vascular endothelial growth factor (VEGF165) cDNA eukaryotic expression vector promoted by HRE, and investigate its targeted inhibiting VEGF expression of osteosarcoma cells in hypoxia environment.

METHODSEukaryotic expression plasmid with HRE promoter was constructed containing luciferase reporter gene and antisense VEGF165 cDNA by using PCR and recombinant DNA techniques. The recombinant vectors were transfected into osteosarcoma cells with lipofectin method. Hypoxia-inducible reporter gene expression was determined by liquid scintillation analyzer and the expression of VEGF protein was detected by ELISA method.

RESULTSThe eukaryotic expression plasmid containing antisense VEGF165 and luciferase promoted by HRE was constructed successfully. After being transferred into MG63 cells, luciferase expression was increased 3.5 x 10(2) times and VEGF protein expression decreased 45% under hypoxia condition.

CONCLUSIONAntisense VEGF165 cDNA expression, efficiently realized by HRE promoter under hypoxia condition, provides an experimental basis for targeted antiangiogenesis of tumors.

Bone Neoplasms ; metabolism ; pathology ; Cell Hypoxia ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1 ; genetics ; Luciferases ; genetics ; metabolism ; Oligodeoxyribonucleotides, Antisense ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Promoter Regions, Genetic ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

ObjectiveTo explore a new model for lens-induced myopia （LIM） in mice and describe the changes of diopter and ocular biological parameters. MethodsTwenty-seven 21-day-old C57BL/6 mice were divided into three groups （ratio， 5：1：3）： LIM group， plano lens （PL） group and normal control （N） group. The right eyes were intervened while the left eyes were left as control. The refraction was detected with retinoscopy after the pupils were dilated with compound topicamide and ocular axial length was measured by optical coherence tomography （OCT） in vivo at baseline and 1， 2， 3， and 4 weeks after the intervention. Paired t test was performed between left and right eyes within each group. Welch's ANOVA was used for comparison among the three groups. When the difference was statistically significant， the Dunnett's T3 was used to correct P value for pairwise comparison. ResultsAfter 2 weeks of defocus induction， the refraction of the intervened eye in LIM group shifted to myopia about （-2.55±1.54） D（t=6.430， P＜0.000 1）， and the ocular axial length （AL） increased about （0.051±0.024） mm（t=7.837， P＜0.000 1）. The difference of interocular change in refraction in LIM group compared with PL group and N group was -2.30 D （P=0.014） and -2.55 D （P＜0.000 1）， respectively. The difference of interocular change in AL in LIM group was 0.048 mm （P<0.000 1） and 0.047 mm （P<0.000 1） compared with that in PL group and N group， respectively. With the extension of intervention time， the degree of myopia drift increased. ConclusionIn this study， a clasp-based and detachable LIM model was described and validated. After 2 weeks of intervention， the refraction shifted significantly toward myopia and the AL increased significantly. The LIM model is simple to construct and can provide a reference for the model construction of animal experiments in myopia research.

OBJECTIVETo practice a more atraumatic, physiological and aesthetically valued approach of construction for neovagina.

METHODSLaparoscopically using peritoneum as neovagina lining.

RESULTSFrom March 2005 to September 2006, this technique was adopted to treat 10 patients whose diagnosis was congenital absence of vagina. The ages of the patients were from 19 to 32. The operation lasted average 2.34 hours. And hospitalization was about 20.5 days. Follow-up ranged from 3 - 12 months. No complication occurred. All of the patients was satisfied with their sexual life.

CONCLUSIONSLaparoscopically assisted neovaginaplasty, in which peritoneum was substituted for vaginal mucous membrane, was a kind of ideal approach of vaginal creation.

Adult ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Peritoneum ; transplantation ; Reconstructive Surgical Procedures ; methods ; Vagina ; abnormalities ; surgery ; Young Adult