HPV16 L2E7 is a fusion protein used for therapeutical vaccine targeting HPV virus. To increase its expression in Escherichia coli, we optimized the codon usage of HPV16 l2e7 gene based on its codon usage bias. The optimized gene of HPV16 sl2e7 was cloned into three different vectors: pGEX-5X-1, pQE30, ET41a, and expressed in JM109, JM109 (DE3) and BL21 (DE3) lines separately. A high expression line was selected with pET41a vector in BL21 (DE3) cells. After optimization of the growth condition, including inoculation amount, IPTG concentration, induction time and temperature, the expression level of HPV16 L2E7 was increased from less than 10% to about 28% of total protein. HPV16 L2E7 protein was then purified from 15 L culture by means of SP Sepharose Fast Flow, Q Sepharose Fast Flow and Superdex 200 pg. After renaturing, HPV16 L2E7 protein with ≥ 95% purity was achieved, which was confirmed via SDS-PAGE gel and Western blotting. The combined use of purified HPV16 L2E7 and CpG helper has shown clear inhibition of tumor growth in mice injected with tumor cells, with six out of eight mice shown no sign of tumor. This study lays a solid foundation for a new pipeline of large-scale vaccine production.
Animals ; Capsid Proteins ; biosynthesis ; Codon ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Human papillomavirus 16 ; Mice ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; biosynthesis ; Papillomavirus E7 Proteins ; biosynthesis ; Papillomavirus Vaccines ; therapeutic use ; Recombinant Fusion Proteins ; biosynthesis

Objective To investigate the predictors of systemic inflammatory response syndrome (SIRS) in patients with acute ischemic stroke. Methods The patients with acute ischemic stroke admitted to hospital from January 2010 to April 2014 were enroled. The demographics, vascular risk factors, baseline clinical data, and laboratory tests in both groups were colected. Flow cytometry was used to analyze the peripheral blood T helper cel (Th) subgroup. Double antibody sandwich enzyme-linked immunosorbent assay was used to detect the levels of peripheral blood interferon-γ (IFN-γ) and interleukin-4 (IL-4). Results A total of 143 patients with acute ischemic stroke were enroled, including 56 in a SIRS group and 87 in a non-SIRS group. Univariate analysis showed that there were significant differences in the history of hypertension, history of stroke, baseline systolic blood pressure, low-density lipoprotein cholesterol level, National Institutes of Health Stroke Scale (NIHSS) score, percentage of Th1 cels, and IFN-γ concentration in patients of both groups (al P < 0. 05). Multivariate logistic regression analysis showed that the NIHSS score ≥6 (odds ratio [ OR] 2. 40, 95% confidence interval [ CI] 1. 24 - 5. 15, P = 0. 008), decreased percentage of Th1 cels (OR 2. 81, 95% CI 1. 51 - 6. 83, P = 0. 013), and decreased IFN-γ concentration (OR 4. 63, 95% CI 1. 01 - 9. 72, P = 0. 004) were the independent predictive factors for occurring SIRS in patients with acute ischemic stroke. Conclusions Severe neurological deficit, decreased percentage of Th1 cels or decreased IFN-γ level may increase the risk of acute ischemic stroke patients with SIRS.

OBJECTIVETo explore a novel non-invasive method in detection of thyroid cancer by Fourier transform infrared (FTIR) spectroscopy.

METHODSSurface FTIR spectra of 15 cases of thyroid cancer and 51 cases of normal subjects were collected. 22 variables of 11 bands including peak positions and relative intensities were measured and all data were statistically analyzed.

RESULTSIn the cancer group: (1) the peak position of 1743 cm(-1) was shifted toward higher wave number (P < 0.05), and that of 1250 cm(-1) to the lower (P < 0.05), when compared to those of normal ones. (2) The relative intensity ratios of I(1546)/I(1460), I(1250)/I(1460), I(1120)/I(1460), I(1080)/I(1460) were significantly increased (P < 0.05). (3) The presence rate of band of 1340 cm(-1) was significantly decreased (P < 0.05).

CONCLUSIONFTIR surface spectra may become a novel powerful non-invasive approach of detecting thyroid cancer in regular routine check-up.

Humans ; Spectroscopy, Fourier Transform Infrared ; methods ; Thyroid Gland ; chemistry ; Thyroid Neoplasms ; chemistry ; diagnosis

OBJECTIVE: To observe the clinical effect of six-step manipulation combined with extracorporeal shock wave in the treatment of knee osteoarthritis. METHODS: Seventy-six patients with KOA from December 2016 to June 2018 were divided into control group and treatment group, 38 in each group. The patients in the control group were treated with oral medicine combined with extracorporeal shock wave therapy, while the patients in the treatment group were treated with six-step manipulation combined with shock wave therapy. The VAS score, WOMAC score and clinical efficacy of the two groups were compared before treatment, 1 day, 1 month and 6 months after treatment. RESULTS: There was no significant difference in VAS score and WOMAC score between the two groups before treatment(>0.05). VAS score and WOMAC score in treatment group were significantly lower than those in control group at 1 day, 1 month and 6 months after treatment, and the difference was statistically significant(<0.05). CONCLUSIONS Six-step manipulation combined with extracorporeal shock wave therapy can significantly alleviate pain and improve knee function in patients with knee osteoarthritis, and the clinical effect is obvious.
Acupuncture Therapy ; Extracorporeal Shockwave Therapy ; Humans ; Knee Joint ; Osteoarthritis, Knee ; therapy ; Treatment Outcome

OBJECTIVETo evaluate the efficacy and safety of capecitabine as first-line therapy in patients with advanced and recurrent colorectal cancer.

METHODSFrom December 2000 to November 2001, sixty patients with advanced and recurrent colorectal cancer received first-line capecitabine treatment given at a dose of 1250 mg/m(2) twice daily, on days 1 - 14 every 21 days. At least 2 cycles were administered.

RESULTSThe overall response rate was 23.3% with 14 PR, 24 SD (40.0%) and 15 PD. The median survival time was 14.7 months. The survival rate was 63.9% at 12-months and 33.4% at 24-months. Grade III-IV adverse effects were diarrhea in 4 patients (6.6%), anemia in 2 (3.3%) and hand-foot syndrome (HFS) in 1 (1.7%); Grade I-II adverse effects were hyperpigmentation in 20 (33.3%), HFS in 18 (30.0%) and diarrhea in 10 (16.7%).

CONCLUSIONCapecitabine is an efficacious and better-tolerated alternative treatment for the patients with advanced and recurrent colorectal cancer.

Adult ; Aged ; Antimetabolites, Antineoplastic ; therapeutic use ; Capecitabine ; Colorectal Neoplasms ; drug therapy ; mortality ; Deoxycytidine ; adverse effects ; analogs & derivatives ; therapeutic use ; Female ; Fluorouracil ; analogs & derivatives ; Humans ; Male ; Middle Aged ; Survival Rate

At present, severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)infection is still rampant worldwide.As of September 10, 2021, there were about 222 million confirmed cases of corona virus disease 2019(COVID-19)and more than 4.6 million deaths worldwide.With the development of COVID-19 vaccines and the gradual vaccination worldwide, the increasing number of cases in children and unvaccinated young people has drawn attention.According to World Health Organization surveillance data, the proportion of COVID-19 infection cases in children gradually increased, and the proportion of cases in the age groups of under 5 years and 5-14 years increased from 1.0% and 2.5% in January 2020 to 2.0% and 8.7% in July 2021, respectively.At present, billions of adults have been vaccinated with various COVID-19 vaccines worldwide, and their protective effects including reducing infection and transmission, reducing severe disease and hospitalization, and reducing death, as well as high safety have been confirmed.Canada, the United States, Europe and other countries have approved the emergency COVID-19 vaccination in children and adolescents aged 12 to 17 years, and China has also approved the phased vaccination of COVID-19 vaccination in children and adolescents aged 3 to 17 years. For smooth advancement and implementation of COVID-19 vaccination in children, academic institutions, including National Clinical Research Center for Respiratory Diseases, National Center for Children′s Health, and The Society of Pediatrics, Chinese Medical Association organized relevant experts to reach this consensus on COVID-19 vaccination in children.

OBJECTIVE: By observing the effect of inner-heating acupuncture on the expression of Wnt1, Axin and β-catenin in the intervertebral disc of rats with lumbar degenerative disease and to explore the regulatory mechanism of inner-heating acupuncture on the apoptosis of annulus fibrosus cells in rats with lumbar degenerative disease. METHODS: Forty SD rats were randomly divided into normal group, model group, inner heating acupuncture group and acupuncture group with 10 rats in each group. Except for normal group, all rats were modeled. Bilateral were selected for inner-heating acupuncture and acupuncture treatment. Western-bolt assay was used to detect the expression of Wnt1, Axin and β-catenin in rat intervertebral discs. RESULTS: Compared with the model group, Wnt1 and β-catenin expression in annulus fibrosus cells of rats in the inner heating acupuncture group and the acupuncture group were significantly decreased(<0.05);the expression of Axin in annulus fibrosus of rats in the inner heating acupuncture group was significantly increased(<0.05). There was no significant difference in the expression of Axin in annulus fibrosus of rats between acupuncture group and model group(>0.05). CONCLUSION Inner heating acupuncture can down regulate the expression of Wnt1 and β-catenin and up regulate the expression of Axin in annulus fibrosus of rats with lumbar degenerative desease. It is suggested that the mechanism may be to regulate the expression of related factors in the β-catenin signaling pathway, so as to achieve the goal of treating lumbar degenerative diseases.
Acupuncture Therapy ; Animals ; Heating ; Intervertebral Disc ; Rats ; Rats, Sprague-Dawley ; beta Catenin

A new neolignan, (-)-(7R,8S,7'E)-3',4-dihydroxy-3-methoxy-8,4'-oxyneoligna-7'-ene-7,9,9'-triol(1), and seven known compounds, 9-(tetrahydropyran-2-yl)-nona-trans,trans-2,8-diene-4,6-diyn-1-ol (2), 9-(tetrahydropyran-2-yl)-trans-non-8-ene-4,6-diyn-1-ol (3), lobetyol (4), lobetyolin (5),dehydrodieoniferyl alcohol (6), 5-hydroxymethylfurfural (7), and 4, 4'-dihydroxy-3, 3'-dimethoxy-trans-stilbene (8), were isolated from the H2O extract of Codonopsis pilosula. The structures of 1-8 were elucidated by spectroscopic methods including NMR, HR-ESI-MS, and CD. In addition, compounds 2 and 3 were isolated from the genus Codonopsis for the first time.

Severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)infection is still worldwide.As a vulnerable group, severe and dead pediatric cases are also reported.Under this severe epidemic situation, children should be well protected.With the widespread vaccination of SARS-CoV-2 vaccine in adults, the infection rate have decreased.Therefore, SARS-CoV-2 vaccine inoculation for children groups step by step is of great significance to the protection of children and the prevention and control of corona virus disease 2019(COVID-19) as a whole.But the safety of children vaccinated with SARS-CoV-2 vaccine is a main concern of parents.Therefore, in order to ensure the safety of vaccination and the implementation of vaccination work, National Clinical Research Center for Respiratory Diseases, National Center for Children′s Health and the Society of Pediatrics, Chinese Medical Association organized experts to interpret the main issue of parents about SARS-CoV-2 vaccine for children, in order to answer the doubts of parents.

Monkeypox is a zoonotic disease.Previous studies have shown that children are vulnerable to monkeypox and are also at high risk for severe disease or complications.In order to improve pediatricians′ understanding of monkeypox and achieve early detection, early diagnosis, early treatment and early disposal, the committee composed of more than 40 experts in the related fields of infectious diseases, pediatrics, infection control and public health formulate this expert consensus, on the basis of the latest clinical management and infection prevention and control for monkeypox released by the World Health Organization (WHO), the guidelines for diagnosis and treatment of monkeypox (version 2022) issued by National Health Commission of the People′s Republic of China and other relevant documents.During the development of this consensus, multidisciplinary experts have repeatedly demonstrated the etiology, epidemiology, transmission, clinical manifestations, laboratory examinations, diagnosis and differential diagnosis, treatment, discharge criteria, prevention, case management process and key points of prevention and control about monkeypox.