1.Cloning and expression of Chlamydia trachomatis heat shock protein 60 gene
Juanhua LIU ; Mukai CHEN ; Qiman LIAO ; Haipian LI ; Yuying TU ; Jiande HAN
Chinese Journal of Dermatology 2009;42(5):318-320
Objective To clone and express Chlamydia trachomatis (Ct) heat shock protein 60 (hsp60) gene. Methods The hsp60 gene fragment was amplified from Ct chromosomal DNA by PCR. After purification and digestion with enzymes Sal I and Not I , the hsp60 gene fragment was inserted into the compatible site of prokaryotic expression vector pET-28a. The constructed recombinant plasmid was identified by PCR, restriction enzyme cleavage and sequencing, then, it was transfected into an expression strain Escherichia coli BL21 (DE3). The expression of fusion protein was induced by isopropy-β-D- thiogalactoside (IPTG) in the host bacteria, and the expressed product was identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blot. Results PCR and restriction enzymes cleavage analysis confirmed that the hsp60 gene was successfully cloned into the recombinant plasmid. DNA sequencing showed that the sequence of cloned gene was fully consistent with the published sequence in Genebank. As revealed by SDS-PAGE, the size of expressed fusion protein approximated 60 kilodaltons, and Western-blot confirmed the expressed product to be the expected protein. The final concentration of fusion protein was 17.85 mg/L with a purity of more than 90%. Conclusions A recombinant expression plasmid pET-28a-hsp60 is successfully constructed in this study, and soluble hsp60 protein is expressed by the recombinant plasmid-transfected E. coli.
2.Efficacy and safety of Tianmeng Oral Liquid in the treatment of hyperprolactinemia caused by antipsychotics in female patients with schizophrenia: a Meta-analysis
Xianjun LAN ; Zhenjuan QIN ; Qiman LIU ; Dongbin CAI ; Yaoyao LU ; Xiong HUANG ; Wei ZHENG
Sichuan Mental Health 2023;36(1):25-31
ObjectiveTo systematically evaluate the efficacy and safety of Tianmeng Oral Liquid in the treatment of hyperprolactinemia caused by antipsychotics in female patients with schizophrenia, and to provide references for intervention of antipsychotic-induced hyperprolactinemia patients. MethodsEnglish databases (PubMed, Cochrane Library, PsycINFO and Embase) and Chinese databases (CNKI and Wanfang) were systematically searched, with a search time frame of the period from database creation to September 16, 2022. Randomized controlled studies (RCTs) on adjunctive Tianmeng Oral Liquid in the treatment of hyperprolactinemia caused by antipsychotics in female patients with schizophrenia were included. Three investigators independently screened the literatures and extracted data according to PICOS principles and evaluated the quality of including literatures, and the Meta-analysis was performed by using RevMan 5.3. ResultsA total of 3 articles including 256 female schizophrenic patients with hyperprolactinemia were included. Meta-analysis results showed that at the treatment endpoint, the improvement of hyperprolactinemia symptoms reported a significant superiority of the intervention group than the control group (RR=1.73, 95% CI: 1.07~2.79, P˂0.05). Similarly, the significant superiority of the intervention group than the control group were also found in reducing serum prolactin levels (WMD=-55.17, 95% CI: -68.16~-42.18, P<0.01) and Positive and Negative Syndrome Scale (PANSS) total scores were lower in the intervention group than in the control group (WMD=-7.36, 95% CI: -8.94~-5.77, P<0.01). ConclusionTianmeng Oral Liquid may help improve clinical efficacy and reduce serum prolactin levels in female schizophrenia patients with hyperprolactinemia. It may improve psychiatric symptoms in female patients with schizophrenia.